Molecular basis for selection and inhibition of HIV-1 escape virus by T cells and KIR2DL2+NK cells
Takayuki Chikata, Kimiko Kuroki, Nozomi Kuse, Anna E. Kliszczak, Wayne Paes, Nanami Tomioka, Robert Parker, Aure Aflalo, Tomohiro Akahoshi, Yu Zhang, Ryoya Yamashita, Ryuma Sakata, Hiroki Kusaka, Yosuke Watanabe, Annalisa Nicastri, Haruki Matsubara, Toyoyuki Ose, Shunsuke Kita, Shinichi Oka, Hiroyuki Gatanaga, Zhansong Lin, Nicola Ternette, Persephone Borrow, Katsumi Maenaka, Masafumi Takiguchi
Nature Communications, 16, 1, Springer Science and Business Media LLC, 06 Nov. 2025, [Peer-reviewed], ［Internationally co-authored］
Scientific journal

Sld3CBD–Cdc45 structural insights into Cdc45 recruitment for CMG complex formation during DNA replication
Hao Li, Izumi Ishizaki, Koji Kato, Xiaomei Sun, Sachiko Muramatsu, Hiroshi Itou, Toyoyuki Ose, Hiroyuki Araki, Min Yao
eLife, 13, eLife Sciences Publications, Ltd, 08 Sep. 2025, [Peer-reviewed]
Scientific journal, DNA replication requires recruitment of Cdc45 and GINS into the MCM double hexamer by initiation factors to form an active helicase, the Cdc45–MCM–GINS (CMG) complex, at the replication origins. The initiation factor Sld3 is a central regulator of Cdc45 and GINS recruitment, working with Sld7 together. However, the mechanism through which Sld3 regulates CMG complex formation remains unclear. Here, we present the structure of the Sld3 Cdc45-binding domain in complex with Cdc45 (Sld3CBD–Cdc45), showing detailed interactions and conformational changes required for binding to each other. The mutant analysis indicated that the binding between Sld3CBD and Cdc45 could be broken easily. We also revealed that Sld3CBD, GINS, and MCM bind to different sites on Cdc45 in the Sld3CBD–CMG model, indicating that after recruitment of Cdc45, Sld7–Sld3 could remain in Cdc45–MCM until CMG formation. The consistency between the particle size of Sld7–Sld3–Cdc45 and the distance between Sld3CBDs in the Cdc45–MCM dimer indicated the binding manner of the Cdc45–Sld3–[Sld7]₂–Sld3–Cdc45 off/on MCM double hexamer. A DNA-binding assay of Sld3 and its complexes with single-stranded ARS1 (autonomously replicating sequence 1) fragments revealed a relationship between the dissociation of Sld7–Sld3 from CMG and the unwound single-stranded DNA. These findings help to further our understanding of the molecular basis of the regulation of CMG complex formation by Sld3.

Solution structure of the C-terminal domain of the measles virus V protein in its free form and mechanistic analysis of STAT2 targeting　　　　　　　　　　　　　　　
Kaho Morita, Nanaka Goda, Madoka Kimoto, Satomi Inaba-Inoue, Nana Yabuno, Aoi Sugiyama, Hiroyuki Kumeta, Toyoyuki Ose
Journal of Virology, Sep. 2025, [Peer-reviewed], [Last author, Corresponding author]

Chain-length preference of trans-acting enoylreductases involved in the biosynthesis of fungal polyhydroxy polyketides
Yuichiro Takekawa, Junya Takino, Shusuke Sato, Hideaki Oikawa, Toyoyuki Ose, Atsushi Minami
Biochemical and Biophysical Research Communications, 761, 151737, 151737, Elsevier BV, May 2025, [Peer-reviewed], [Corresponding author]
Scientific journal

Structural analysis reveals how tetrameric tyrosine-phosphorylated STAT1 is targeted by the rabies virus P-protein
Aoi Sugiyama, Miku Minami, Kaito Ugajin, Satomi Inaba-Inoue, Nana Yabuno, Yuichiro Takekawa, Sun Xiaomei, Shiho Takei, Mina Sasaki, Tomo Nomai, Xinxin Jiang, Shunsuke Kita, Katsumi Maenaka, Mika Hirose, Min Yao, Paul R. Gooley, Gregory W. Moseley, Yukihiko Sugita, Toyoyuki Ose
Science Signaling, 18, 878, American Association for the Advancement of Science (AAAS), 18 Mar. 2025, [Peer-reviewed], [Last author, Corresponding author], ［Internationally co-authored］
Scientific journal, Signal transducer and activator of transcription (STAT) family members mediate signaling in the Janus kinase (JAK)–STAT pathway and are activated by phosphorylation at a conserved tyrosine residue, resulting in dimerization through reciprocal interactions between the phosphotyrosine and a Src homology 2 (SH2) domain. Tyrosine-phosphorylated STAT (pY-STAT) then translocates to the nucleus to induce the expression of genes encoding antiviral proteins. Although the active and functional forms of STATs are conventionally considered to be dimers, STATs can undergo higher-order oligomerization, which is implicated in regulating transcriptional activity. We present the cryo–electron microscopy (cryo-EM) structure of the tetrameric form of intact pY-STAT1 in complex with DNA, which indicates that interactions between the amino-terminal domains (NTDs) of STAT1 induce oligomerization. The tetrameric structure revealed a compact conformation with a previously uncharacterized binding interface: Two DNA-bound dimers are twofold symmetrically aligned to transform into a tandem DNA-binding model without NTD dimer separation. Moreover, biochemical analyses indicated that the rabies virus P-protein selectively targeted tetrameric pY-STAT1. Combined with data showing which regions contribute to the interaction between pY-STAT1 and the P-protein, we constructed a binding model explaining how P recognizes the pY-STAT1 tetramer. These data provide insight into how pathogenic viruses target signaling pathways that mediate the host immune response.

Crystallographic analysis of the Escherichia coli tRNA seleno-modification enzyme in complex with tRNA
Takuya Usui, Sayaka Ono, Akiyoshi Nakamura, Koji Kato, Toyoyuki Ose, Min Yao
Acta Crystallographica Section F Structural Biology Communications, 81, 2, International Union of Crystallography (IUCr), 09 Jan. 2025, [Peer-reviewed]
Scientific journal, The bacterial enzyme tRNA 2-selenouridine synthase (SelU) catalyzes the conversion of 5-substituted 2-thiouridine (R5S2U) to 5-substituted 2-selenouridine (R5Se2U) at the wobble positions of several tRNAs. Seleno-modification potentially regulates translation efficiency in response to selenium availability. Notably, SelU uses the 2-geranylthiouridine (R5geS2U) intermediate for sulfur removal, and this geranylthiol (geS) is a unique leaving group among tRNA-maturation enzymes. However, the underlying sequence of the SelU reaction remains unclear. Here, a crystallographic study of the Escherichia coli SelU–tRNA complex is reported. Robust and well formed SelU–tRNA crystals were obtained after several optimizations, including co-expression with tRNA and additive screening. Diffraction data were collected at a resolution of 3.10 Å using a wavelength of 1.0000 Å. The crystals belonged to space group C2, and the phase was determined by molecular replacement using the AlphaFold2-predicted SelU structure as a search model. Electron-density mapping revealed the presence of two SelU–tRNA complexes in the asymmetric unit.

Molecular mechanism for the substrate specificity of Arthrobacter globiformis M6 α-glucosidase CmmB, belonging to glycoside hydrolase family 13 subfamily 30
Wataru Saburi, Takayoshi Tagami, Takuya Usui, Jian Yu, Toyoyuki Ose, Min Yao, Haruhide Mori
Food Bioscience, 61, 104516, 104516, Elsevier BV, Oct. 2024, [Peer-reviewed]
Scientific journal

Glycan-shielded homodimer structure and dynamical features of the canine distemper virus hemagglutinin relevant for viral entry and efficient vaccination.
Hideo Fukuhara, Kohei Yumoto, Miyuki Sako, Mizuho Kajikawa, Toyoyuki Ose, Mihiro Kawamura, Mei Yoda, Surui Chen, Yuri Ito, Shin Takeda, Mwila Mwaba, Jiaqi Wang, Takao Hashiguchi, Jun Kamishikiryo, Nobuo Maita, Chihiro Kitatsuji, Makoto Takeda, Kimiko Kuroki, Katsumi Maenaka
eLife, 12, 24 Jul. 2024, [Peer-reviewed], [International Magazine]
English, Scientific journal, Canine distemper virus (CDV) belongs to morbillivirus, including measles virus (MeV) and rinderpest virus, which causes serious immunological and neurological disorders in carnivores, including dogs and rhesus monkeys, as recently reported, but their vaccines are highly effective. The attachment glycoprotein hemagglutinin (CDV-H) at the CDV surface utilizes signaling lymphocyte activation molecule (SLAM) and Nectin-4 (also called poliovirus-receptor-like-4; PVRL4) as entry receptors. Although fusion models have been proposed, the molecular mechanism of morbillivirus fusion entry is poorly understood. Here, we determined the crystal structure of the globular head domain of CDV-H vaccine strain at 3.2 Å resolution, revealing that CDV-H exhibits a highly tilted homodimeric form with a six-bladed β-propeller fold. While the predicted Nectin-4-binding site is well conserved with that of MeV-H, that of SLAM is similar but partially different, which is expected to contribute to host specificity. Five N-linked sugars covered a broad area of the CDV-H surface to expose receptor-binding sites only, supporting the effective production of neutralizing antibodies. These features are common to MeV-H, although the glycosylation sites are completely different. Furthermore, real-time observation using high-speed atomic force microscopy revealed highly mobile features of the CDV-H dimeric head via the connector region. These results suggest that sugar-shielded tilted homodimeric structure and dynamic conformational changes are common characteristics of morbilliviruses and ensure effective fusion entry and vaccination.

STAP-2–Derived Peptide Suppresses TCR-Mediated Signals to Initiate Immune Responses
Yuto Sasaki, Kodai Saitoh, Kota Kagohashi, Toyoyuki Ose, Shoya Kawahara, Yuichi Kitai, Ryuta Muromoto, Yuichi Sekine, Michiko Ichii, Akihiko Yoshimura, Kenji Oritani, Jun-ichi Kashiwakura, Tadashi Matsuda
The Journal of Immunology, The American Association of Immunologists, 07 Jul. 2023, [Peer-reviewed]
Scientific journal, Abstract

Signal-transducing adaptor protein-2 (STAP-2) is an adaptor protein that contains pleckstrin and Src homology 2–like domains, as well as a proline-rich region in its C-terminal region. Our previous study demonstrated that STAP-2 positively regulates TCR signaling by associating with TCR-proximal CD3ζ ITAMs and the lymphocyte-specific protein tyrosine kinase. In this study, we identify the STAP-2 interacting regions of CD3ζ ITAMs and show that the STAP-2–derived synthetic peptide (iSP2) directly interacts with the ITAM sequence and blocks the interactions between STAP-2 and CD3ζ ITAMs. Cell-penetrating iSP2 was delivered into human and murine T cells. iSP2 suppressed cell proliferation and TCR-induced IL-2 production. Importantly, iSP2 treatment suppressed TCR-mediated activation of naive CD4+ T cells and decreased immune responses in CD4+ T cell–mediated experimental autoimmune encephalomyelitis. It is likely that iSP2 is a novel immunomodulatory tool that modulates STAP-2-mediated activation of TCR signaling and represses the progression of autoimmune diseases.

Structural insights into the substrate specificity and activity of a novel mannose 2-epimerase from Runella slithyformis.
Hang Wang, Xiaomei Sun, Wataru Saburi, Saki Hashiguchi, Jian Yu, Toyoyuki Ose, Haruhide Mori, Min Yao
Acta crystallographica. Section D, Structural biology, 79, Pt 7, 585, 595, 01 Jul. 2023, [Peer-reviewed], [International Magazine]
English, Scientific journal, Mannose 2-epimerase (ME), a member of the acylglucosamine 2-epimerase (AGE) superfamily that catalyzes epimerization of D-mannose and D-glucose, has recently been characterized to have potential for D-mannose production. However, the substrate-recognition and catalytic mechanism of ME remains unknown. In this study, structures of Runella slithyformis ME (RsME) and its D254A mutant [RsME(D254A)] were determined in their apo forms and as intermediate-analog complexes [RsME-D-glucitol and RsME(D254A)-D-glucitol]. RsME possesses the (α/α)6-barrel of the AGE superfamily members but has a unique pocket-covering long loop (loopα7-α8). The RsME-D-glucitol structure showed that loopα7-α8 moves towards D-glucitol and closes the active pocket. Trp251 and Asp254 in loopα7-α8 are only conserved in MEs and interact with D-glucitol. Kinetic analyses of the mutants confirmed the importance of these residues for RsME activity. Moreover, the structures of RsME(D254A) and RsME(D254A)-D-glucitol revealed that Asp254 is vital for binding the ligand in a correct conformation and for active-pocket closure. Docking calculations and structural comparison with other 2-epimerases show that the longer loopα7-α8 in RsME causes steric hindrance upon binding to disaccharides. A detailed substrate-recognition and catalytic mechanism for monosaccharide-specific epimerization in RsME has been proposed.

Quick and Spontaneous Transformation between [3Fe–4S] and [4Fe–4S] Iron–Sulfur Clusters in the tRNA-Thiolation Enzyme TtuA
Masato Ishizaka, Minghao Chen, Shun Narai, Yoshikazu Tanaka, Toyoyuki Ose, Masaki Horitani, Min Yao
International Journal of Molecular Sciences, 24, 1, 833, 833, MDPI AG, 03 Jan. 2023, [Peer-reviewed]
Scientific journal, Iron–sulfur (Fe–S) clusters are essential cofactors for enzyme activity. These Fe–S clusters are present in structurally diverse forms, including [4Fe–4S] and [3Fe–4S]. Type-identification of the Fe–S cluster is indispensable in understanding the catalytic mechanism of enzymes. However, identifying [4Fe–4S] and [3Fe–4S] clusters in particular is challenging because of their rapid transformation in response to oxidation–reduction events. In this study, we focused on the relationship between the Fe–S cluster type and the catalytic activity of a tRNA-thiolation enzyme (TtuA). We reconstituted [4Fe–4S]-TtuA, prepared [3Fe–4S]-TtuA by oxidizing [4Fe–4S]-TtuA under strictly anaerobic conditions, and then observed changes in the Fe–S clusters in the samples and the enzymatic activity in the time-course experiments. Electron paramagnetic resonance analysis revealed that [3Fe–4S]-TtuA spontaneously transforms into [4Fe–4S]-TtuA in minutes to one hour without an additional free Fe source in the solution. Although the TtuA immediately after oxidation of [4Fe–4S]-TtuA was inactive [3Fe–4S]-TtuA, its activity recovered to a significant level compared to [4Fe–4S]-TtuA after one hour, corresponding to an increase of [4Fe–4S]-TtuA in the solution. Our findings reveal that [3Fe–4S]-TtuA is highly inactive and unstable. Moreover, time-course analysis of structural changes and activity under strictly anaerobic conditions further unraveled the Fe–S cluster type used by the tRNA-thiolation enzyme.

Structural and molecular insight into antibody recognition of dynamic neoepitopes in membrane tethered MUC1 of pancreatic cancer cells and secreted exosomes
Hajime Wakui, Yasuhiro Yokoi, Chieko Horidome, Toyoyuki Ose, Min Yao, Yoshikazu Tanaka, Hiroshi Hinou, Shin-Ichiro Nishimura
RSC Chemical Biology, Royal Society of Chemistry (RSC), 2023, [Peer-reviewed]
Scientific journal, We unveil for the first time that pancreatic cancer cells (PANC-1) and secreted exosomes express MUC1 bearing cancer-relevant dynamic epitopes recognized specifically by an anti-MUC1 antibody (SN-131).

A peptide derived from adaptor protein STAP-2 inhibits tumor progression by downregulating epidermal growth factor receptor signaling
Taiga Maemoto, Yuichi Kitai, Haruka Shoji, Runa Takahashi, Shunsuke Yamada, Shiho Takei, Daiki Ito, Ryuta Muromoto, Jun-ichi Kashiwakura, Haruka Handa, Ari Hashimoto, Shigeru Hashimoto, Toyoyuki Ose, Kenji Oritani, Tadashi Matsuda
Journal of Biological Chemistry, in press, 299, 1, 102724, 102724, Dec. 2022, [Peer-reviewed], [International Magazine]
English, Scientific journal, Signal-transducing adaptor family member-2 (STAP-2) is an adaptor protein that regulates various intracellular signals. We previously demonstrated that STAP-2 binds to epidermal growth factor receptor (EGFR) and facilitates its stability and activation of EGFR signaling in prostate cancer cells. Inhibition of this interaction may be a promising direction for cancer treatment. Here, we found that 2D5 peptide, a STAP-2-derived peptide, blocked STAP-2-EGFR interactions and suppressed EGFR-mediated proliferation in several cancer cell lines. 2D5 peptide inhibited tumor growth of human prostate cancer cell line DU145 and human lung cancer cell line A549 in murine xenograft models. Additionally, we determined that EGFR signaling and its stability were decreased by 2D5 peptide treatment during EGF stimulation. In conclusion, our study shows that 2D5 peptide is a novel anti-cancer peptide that inhibits STAP-2-mediated activation of EGFR signaling and suppresses prostate and lung cancer progression.

Importance of accessibility to the extracellular juxtamembrane stalk region of membrane protein for substrate recognition by viral ubiquitin ligase K5.
Mizuho Kajikawa, Mizuki Hata, Maho Ishimura, Nanae Imaizumi, Minako Kimura, Kei Miyano, Toyoyuki Ose, Daisuke Asai, Satoshi Ishido, Taisei Kanamoto
The Biochemical journal, 479, 20, 2261, 2278, 28 Oct. 2022, [Peer-reviewed], [International Magazine]
English, Scientific journal, Kaposi's sarcoma-associated herpesvirus (KSHV) is a carcinogenic virus that latently infects B cells and causes malignant tumors in immunocompromised patients. KSHV utilizes two viral E3 ubiquitin ligases, K3 and K5, in KSHV-infected cells to mediate the polyubiquitination-dependent down-regulation of several host membrane proteins involved in the immune system. Although K3 and K5 are members of the same family and have similar structural topologies, K3 and K5 have different substrate specificities. Hence, K5 may have a different substrate recognition mode than K3; however, the molecular basis of substrate recognition remains unclear. Here, we investigated the reason why human CD8α, which is known not to be a substrate for both K3 and K5, is not recognized by them, to obtain an understanding for molecular basis of substrate specificity. CD8α forms a disulfide-linked homodimer under experimental conditions to evaluate the viral ligase-mediated down-regulation. It is known that two interchain disulfide linkages in the stalk region between each CD8α monomer (Cys164-Cys164 and Cys181-Cys181) mediate homodimerization. When the interchain disulfide linkage of Cys181-Cys181 was eliminated, CD8α was down-regulated by K5 with a functional RING variant (RINGv) domain via polyubiquitination at the cytoplasmic tail. Aspartic acid, located at the stalk/transmembrane interface of CD8α, was essential for K5-mediated down-regulation of the CD8α mutant without a Cys181-Cys181 linkage. These results suggest that disulfide linkage near the stalk/transmembrane interface critically inhibits substrate targeting by K5. Accessibility to the extracellular juxtamembrane stalk region of membrane proteins may be important for substrate recognition by the viral ubiquitin ligase K5.

Characterization of Single-Chain Fv Fragments of Neutralizing Antibodies to Rabies Virus Glycoprotein.
Kohei Yumoto, Tomoaki Arisaka, Kazuma Okada, Kyosuke Aoki, Toyoyuki Ose, Tatsunori Masatani, Makoto Sugiyama, Naoto Ito, Hideo Fukuhara, Katsumi Maenaka
Viruses, 13, 11, 19 Nov. 2021, [Peer-reviewed], [International Magazine]
English, Scientific journal, Rabies has almost a 100% case-fatality rate and kills more than 59,000 people annually around the world. There is no established treatment for rabies. The rabies virus (RABV) expresses only the glycoprotein (RABVG) at the viral surface, and it is the target for the neutralizing antibodies. We previously established mouse monoclonal antibodies, 15-13 and 12-22, which showed neutralizing activity against the RABV, targeting the sequential and conformational epitopes on the RABVG, respectively. However, the molecular basis for the neutralizing activity of these antibodies is not yet fully understood. In this study, we evaluated the binding characteristics of the Fab fragments of the 15-13 and 12-22 antibodies. The recombinant RABVG protein, in prefusion form for the binding analysis, was prepared by the silkworm-baculovirus expression system. Biolayer interferometry (BLI) analysis indicated that the 15-13 Fab interacts with the RABVG, with a KD value at the nM level, and that the 12-22 Fab has a weaker binding affinity (KD ~ μM) with the RABVG compared to the 15-13 Fab. Furthermore, we determined the amino acid sequences of both the antibodies and the designed single-chain Fv fragments (scFvs) of the 15-13 and 12-22 antibodies as another potential biopharmaceutical for targeting rabies. The 15-13 and 12-22 scFvs were successfully prepared by the refolding method and were shown to interact with the RABVG at the nM level and the μM level of the KD, respectively. These binding characteristics were similar to that of each Fab. On the other hand, differential scanning fluorometry (DSF) revealed that the thermal stability of these scFvs decreases compared to their Fabs. While the improvement of the stability of scFvs will still be required, these results provide insights into the neutralizing activity and the potential therapeutic use of antibody fragments for RABV infection.

Structural comparison of the C-terminal domain of functionally divergent lyssavirus P proteins.
Aoi Sugiyama, Tomo Nomai, Xinxin Jiang, Miku Minami, Min Yao, Katsumi Maenaka, Naoto Ito, Paul R Gooley, Gregory W Moseley, Toyoyuki Ose
Biochemical and biophysical research communications, 529, 2, 507, 512, 20 Aug. 2020, [Peer-reviewed], [Last author, Corresponding author], [International Magazine]
English, Scientific journal, Lyssavirus P protein is a multifunctional protein that interacts with numerous host-cell proteins. The C-terminal domain (CTD) of P is important for inhibition of JAK-STAT signaling enabling the virus to evade host immunity. Several regions on the surface of rabies virus P are reported to interact with host factors. Among them, an extended, discrete hydrophobic patch of P CTD is notable. Although structures of P CTD of two strains of rabies virus, and of mokola virus have been solved, the structure of P CTD for Duvenhage virus, which is functionally divergent from these species for immune evasion function, is not known. Here, we analyze the structures of P CTD of Duvenhage and of a distinct rabies virus strain to gain further insight on the nature and potential function of the hydrophobic surface. Molecular contacts in crystals suggest that the hydrophobic patch is important to intermolecular interactions with other proteins, which differ between the lyssavirus species.

The Measles Virus V Protein Binding Site to STAT2 Overlaps That of IRF9.
Yuma Nagano, Aoi Sugiyama, Madoka Kimoto, Takuya Wakahara, Yasuyo Noguchi, Xinxin Jiang, Shinya Saijo, Nobutaka Shimizu, Nana Yabuno, Min Yao, Paul R Gooley, Gregory W Moseley, Takashi Tadokoro, Katsumi Maenaka, Toyoyuki Ose
Journal of virology, 94, 17, 17 Aug. 2020, [Peer-reviewed], [Last author, Corresponding author], [International Magazine]
English, Scientific journal, Measles virus (MeV) is a highly immunotropic and contagious pathogen that can even diminish preexisting antibodies and remains a major cause of childhood morbidity and mortality worldwide despite the availability of effective vaccines. MeV is one of the most extensively studied viruses with respect to the mechanisms of JAK-STAT antagonism. Of the three proteins translated from the MeV P gene, P and V are essential for inactivation of this pathway. However, the lack of data from direct analyses of the underlying interactions means that the detailed molecular mechanism of antagonism remains unresolved. Here, we prepared recombinant MeV V protein, which is responsible for human JAK-STAT antagonism, and a panel of variants, enabling the biophysical characterization of V protein, including direct V/STAT1 and V/STAT2 interaction assays. Unambiguous direct interactions between the host and viral factors, in the absence of other factors such as Jak1 or Tyk2, were observed, and the dissociation constants were quantified for the first time. Our data indicate that interactions between the C-terminal region of V and STAT2 is 1 order of magnitude stronger than that of the N-terminal region of V and STAT1. We also clarified that these interactions are completely independent of each other. Moreover, results of size exclusion chromatography demonstrated that addition of MeV-V displaces STAT2-core, a rigid region of STAT2 lacking the N- and C-terminal domains, from preformed complexes of STAT2-core/IRF-associated domain (IRF9). These results provide a novel model whereby MeV-V can not only inhibit the STAT2/IRF9 interaction but also disrupt preassembled interferon-stimulated gene factor 3.IMPORTANCE To evade host immunity, many pathogenic viruses inactivate host Janus kinase signal transducer and activator of transcription (STAT) signaling pathways using diverse strategies. Measles virus utilizes P and V proteins to counteract this signaling pathway. Data derived largely from cell-based assays have indicated several amino acid residues of P and V proteins as important. However, biophysical properties of V protein or its direct interaction with STAT molecules using purified proteins have not been studied. We have developed novel molecular tools enabling us to identify a novel molecular mechanism for immune evasion whereby V protein disrupts critical immune complexes, providing a clear strategy by which measles virus can suppress interferon-mediated antiviral gene expression.

Efficient preparation of human and mouse CD1d proteins using silkworm baculovirus expression system.
Hiroki Kusaka, Shunsuke Kita, Takashi Tadokoro, Kouki Yoshida, Yoshiyuki Kasai, Harumi Niiyama, Yukari Fujimoto, Shinya Hanashima, Michio Murata, Shigeru Sugiyama, Toyoyuki Ose, Kimiko Kuroki, Katsumi Maenaka
Protein expression and purification, 172, 105631, 105631, Aug. 2020, [Peer-reviewed], [International Magazine]
English, Scientific journal, CD1d is a major histocompatibility complex (MHC) class I-like glycoprotein and binds to glycolipid antigens that are recognized by natural killer T (NKT) cells. To date, our understanding of the structural basis for glycolipid binding and receptor recognition of CD1d is still limited. Here, we established a preparation method for the ectodomain of human and mouse CD1d using a silkworm-baculovirus expression system. The co-expression of human and mouse CD1d and β2-microglobulin (β2m) in the silkworm-baculovirus system was successful, but the yield of human CD1d was low. A construct of human CD1d fused with β2m via a flexible GS linker as a single polypeptide was prepared to improve protein yield. The production of this single-chained complex was higher (50 μg/larva) than that of the co-expression complex. Furthermore, differential scanning calorimetry revealed that the linker made the CD1d complex more stable and homogenous. These results suggest that the silkworm-baculovirus expression system is useful for structural and biophysical studies of CD1d in several aspects including low cost, easy handling, biohazard-free, rapid, and high yielding.

A straightforward approach to antibodies recognising cancer specific glycopeptidic neoepitopes
Hajime Wakui, Yoshikazu Tanaka, Toyoyuki Ose, Isamu Matsumoto, Koji Kato, Yao Min, Taro Tachibana, Masaharu Sato, Kentaro Naruchi, Fayna Garcia Martin, Hiroshi Hinou, Shin Ichiro Nishimura
Chemical Science, 11, 19, 4999, 5006, 21 May 2020, [Peer-reviewed]
English, Scientific journal

Structure of HIV-2 Nef Reveals Features Distinct from HIV-1 Involved in Immune Regulation.
Kengo Hirao, Sophie Andrews, Kimiko Kuroki, Hiroki Kusaka, Takashi Tadokoro, Shunsuke Kita, Toyoyuki Ose, Sarah L Rowland-Jones, Katsumi Maenaka
iScience, 23, 1, 100758, 100758, 24 Jan. 2020, [Peer-reviewed], [International Magazine]
English, The human immunodeficiency virus (HIV) accessory protein Nef plays a major role in establishing and maintaining infection, particularly through immune evasion. Many HIV-2-infected people experience long-term viral control and survival, resembling HIV-1 elite control. HIV-2 Nef has overlapping but also distinct functions from HIV-1 Nef. Here we report the crystal structure of HIV-2 Nef core. The di-leucine sorting motif forms a helix bound to neighboring molecules, and moreover, isothermal titration calorimetry demonstrated that the CD3 endocytosis motif can directly bind to HIV-2 Nef, ensuring AP-2-mediated endocytosis for CD3. The highly conserved C-terminal region forms a α-helix, absent from HIV-1. We further determined the structure of simian immunodeficiency virus (SIV) Nef harboring this region, demonstrating similar C-terminal α-helix, which may contribute to AP-1 binding for MHC-I downregulation. These results provide insights into the distinct pathogenesis of HIV-2 infection.

Biophysical characterization and single-chain Fv construction of a neutralizing antibody to measles virus.
Takashi Tadokoro, Mst Lubna Jahan, Yuri Ito, Maino Tahara, Surui Chen, Atsutoshi Imai, Natsumi Sugimura, Koki Yoshida, Mizuki Saito, Toyoyuki Ose, Takao Hashiguchi, Makoto Takeda, Hideo Fukuhara, Katsumi Maenaka
The FEBS journal, 287, 1, 145, 159, Jan. 2020, [Peer-reviewed], [International Magazine]
English, The measles virus (MV) is a major cause of childhood morbidity and mortality worldwide. We previously established a mouse monoclonal antibody, 2F4, which shows high neutralizing titers against eight different genotypes of MV. However, the molecular basis for the neutralizing activity of the 2F4 antibody remains incompletely understood. Here, we have evaluated the binding characteristics of a Fab fragment of the 2F4 antibody. Using the MV infectious assay, we demonstrated that 2F4 Fab inhibits viral entry via either of two cellular receptors, SLAM and Nectin4. Surface plasmon resonance (SPR) analysis of recombinant proteins indicated that 2F4 Fab interacts with MV hemagglutinin (MV-H) with a KD value at the nm level. Furthermore, we designed a single-chain Fv fragment of 2F4 antibody as another potential biopharmaceutical to target measles. The stable 2F4 scFv was successfully prepared by the refolding method and shown to interact with MV-H at the μm level. Like 2F4 Fab, scFv inhibited receptor binding and viral entry. This indicates that 2F4 mAb uses the receptor-binding site and/or a neighboring region as an epitope with high affinity. These results provide insight into the neutralizing activity and potential therapeutic use of antibody fragments for MV infection.

Structural and Functional Basis for LILRB Immune Checkpoint Receptor Recognition of HLA-G Isoforms.
Kimiko Kuroki, Haruki Matsubara, Ryo Kanda, Naoyuki Miyashita, Mitsunori Shiroishi, Yuko Fukunaga, Jun Kamishikiryo, Atsushi Fukunaga, Hideo Fukuhara, Kaoru Hirose, Joan S Hunt, Yuji Sugita, Shunsuke Kita, Toyoyuki Ose, Katsumi Maenaka
Journal of immunology (Baltimore, Md. : 1950), 203, 12, 3386, 3394, 15 Dec. 2019, [Peer-reviewed], [International Magazine]
English, Scientific journal, Human leukocyte Ig-like receptors (LILR) LILRB1 and LILRB2 are immune checkpoint receptors that regulate a wide range of physiological responses by binding to diverse ligands, including HLA-G. HLA-G is exclusively expressed in the placenta, some immunoregulatory cells, and tumors and has several unique isoforms. However, the recognition of HLA-G isoforms by LILRs is poorly understood. In this study, we characterized LILR binding to the β2-microglobulin (β2m)-free HLA-G1 isoform, which is synthesized by placental trophoblast cells and tends to dimerize and multimerize. The multimerized β2m-free HLA-G1 dimer lacked detectable affinity for LILRB1, but bound strongly to LILRB2. We also determined the crystal structure of the LILRB1 and HLA-G1 complex, which adopted the typical structure of a classical HLA class I complex. LILRB1 exhibits flexible binding modes with the α3 domain, but maintains tight contacts with β2m, thus accounting for β2m-dependent binding. Notably, both LILRB1 and B2 are oriented at suitable angles to permit efficient signaling upon complex formation with HLA-G1 dimers. These structural and functional features of ligand recognition by LILRs provide novel insights into their important roles in the biological regulations.

Structural Elucidation of Viral Antagonism of Innate Immunity at the STAT1 Interface.
Hossain MA, Larrous F, Rawlinson SM, Zhan J, Sethi A, Ibrahim Y, Aloi M, Lieu KG, Mok YF, Griffin MDW, Ito N, Ose T, Bourhy H, Moseley GW, Gooley PR
Cell reports, 29, 7, 1934, 1945.e8, Nov. 2019, [Peer-reviewed]

Specificity of Morbillivirus Hemagglutinins to Recognize SLAM of Different Species.
Hideo Fukuhara, Yuri Ito, Miyuki Sako, Mizuho Kajikawa, Koki Yoshida, Fumio Seki, Mwila Hilton Mwaba, Takao Hashiguchi, Masa-Aki Higashibata, Toyoyuki Ose, Kimiko Kuroki, Makoto Takeda, Katsumi Maenaka
Viruses, 11, 8, 19 Aug. 2019, [Peer-reviewed], [International Magazine]
English, Measles virus (MV) and canine distemper virus (CDV) are highly contagious and deadly, forming part of the morbillivirus genus. The receptor recognition by morbillivirus hemagglutinin (H) is important for determining tissue tropism and host range. Recent reports largely urge caution as regards to the potential expansion of host specificities of morbilliviruses. Nonetheless, the receptor-binding potential in different species of morbillivirus H proteins is largely unknown. Herein, we show that the CDV-H protein binds to the dog signaling lymphocyte activation molecule (SLAM), but not to the human, tamarin, or mouse SLAM. In contrast, MV-H can bind to human, tamarin and dog SLAM, but not to that of mice. Notably, MV binding to dog SLAM showed a lower affinity and faster kinetics than that of human SLAM, and MV exhibits a similar entry activity in dog SLAM- and human SLAM-expressing Vero cells. The mutagenesis study using a fusion assay, based on the MV-H-SLAM complex structure, revealed differences in tolerance for the receptor specificity between MV-H and CDV-H. These results provide insights into H-SLAM specificity related to potential host expansion.

¹H, ¹⁵N and ¹³C resonance assignments of the C-terminal domain of the P protein of the Nishigahara strain of rabies virus.
Zhan J, Hossain MA, Sethi A, Ose T, Moseley GW, Gooley PR
Biomolecular NMR assignments, 13, 1, 5, 8, Apr. 2019, [Peer-reviewed]

Neutron crystallographic study of heterotrimeric glutamine amidotransferase CAB.
Li L, Adachi M, Yu J, Kato K, Shinoda A, Ostermann A, Schrader TE, Ose T, Yao M
Acta crystallographica. Section F, Structural biology communications, 75, Pt 3, 193, 196, Mar. 2019, [Peer-reviewed]

Crystal structure of the complex between venom toxin and serum inhibitor from Viperidae snake.
Narumi Shioi, Takashi Tadokoro, Seijiro Shioi, Yuki Okabe, Haruki Matsubara, Shunsuke Kita, Toyoyuki Ose, Kimiko Kuroki, Shigeyuki Terada, Katsumi Maenaka
The Journal of biological chemistry, 294, 4, 1250, 1256, 25 Jan. 2019, [Peer-reviewed], [International Magazine]
English, Venomous snakes have endogenous proteins that neutralize the toxicity of their venom components. We previously identified five small serum proteins (SSP-1-SSP-5) from a highly venomous snake belonging to the family Viperidae as inhibitors of various toxins from snake venom. The endogenous inhibitors belong to the prostate secretory protein of 94 amino acids (PSP94) family. SSP-2 interacts with triflin, which is a member of the cysteine-rich secretory protein (CRISP) family that blocks smooth muscle contraction. However, the structural basis for the interaction and the biological roles of these inhibitors are largely unknown. Here, we determined the crystal structure of the SSP-2-triflin complex at 2.3 Å resolution. A concave region centrally located in the N-terminal domain of triflin is fully occupied by the terminal β-strands of SSP-2. SSP-2 does not bind tightly to the C-terminal cysteine-rich domain of triflin; this domain is thought to be responsible for its channel-blocker function. Instead, the cysteine-rich domain is tilted 7.7° upon binding to SSP-2, and the inhibitor appears to sterically hinder triflin binding to calcium channels. These results help explain how an endogenous inhibitor prevents the venomous protein from maintaining homeostasis in the host. Furthermore, this interaction also sheds light on the binding interface between the human homologues PSP94 and CRISP-3, which are up-regulated in prostate and ovarian cancers.

Structure of MHC class I-like MILL2 reveals heparan-sulfate binding and interdomain flexibility.
Mizuho Kajikawa, Toyoyuki Ose, Yuko Fukunaga, Yuki Okabe, Naoki Matsumoto, Kento Yonezawa, Nobutaka Shimizu, Simon Kollnberger, Masanori Kasahara, Katsumi Maenaka
Nature communications, 9, 1, 4330, 4330, 18 Oct. 2018, [Peer-reviewed], [International Magazine]
English, The MILL family, composed of MILL1 and MILL2, is a group of nonclassical MHC class I molecules that occur in some orders of mammals. It has been reported that mouse MILL2 is involved in wound healing; however, the molecular mechanisms remain unknown. Here, we determine the crystal structure of MILL2 at 2.15 Å resolution, revealing an organization similar to classical MHC class I. However, the α1-α2 domains are not tightly fixed on the α3-β2m domains, indicating unusual interdomain flexibility. The groove between the two helices in the α1-α2 domains is too narrow to permit ligand binding. Notably, an unusual basic patch on the α3 domain is involved in the binding to heparan sulfate which is essential for MILL2 interactions with fibroblasts. These findings suggest that MILL2 has a unique structural architecture and physiological role, with binding to heparan sulfate proteoglycans on fibroblasts possibly regulating cellular recruitment in biological events.

A Sialylated Voltage-Dependent Ca2+ Channel Binds Hemagglutinin and Mediates Influenza A Virus Entry into Mammalian Cells.
Yoichiro Fujioka, Shinya Nishide, Toyoyuki Ose, Tadaki Suzuki, Izumi Kato, Hideo Fukuhara, Mari Fujioka, Kosui Horiuchi, Aya O Satoh, Prabha Nepal, Sayaka Kashiwagi, Jing Wang, Mika Horiguchi, Yuko Sato, Sarad Paudel, Asuka Nanbo, Tadaaki Miyazaki, Hideki Hasegawa, Katsumi Maenaka, Yusuke Ohba
Cell host & microbe, 23, 6, 809, 818, 13 Jun. 2018, [Peer-reviewed], [International Magazine]
English, Scientific journal, Influenza A virus (IAV) infection is initiated by the attachment of the viral glycoprotein hemagglutinin (HA) to sialic acid on the host cell surface. However, the sialic acid-containing receptor crucial for IAV infection has remained unidentified. Here, we show that HA binds to the voltage-dependent Ca2+ channel Cav1.2 to trigger intracellular Ca2+ oscillations and subsequent IAV entry and replication. IAV entry was inhibited by Ca2+ channel blockers (CCBs) or by knockdown of Cav1.2. The CCB diltiazem also inhibited virus replication in vivo. Reintroduction of wild-type but not the glycosylation-deficient mutants of Cav1.2 restored Ca2+ oscillations and virus infection in Cav1.2-depleted cells, demonstrating the significance of Cav1.2 sialylation. Taken together, we identify Cav1.2 as a sialylated host cell surface receptor that binds HA and is critical for IAV entry.

The Role of Heparan Sulfate Proteoglycans as an Attachment Factor for Rabies Virus Entry and Infection.
Sasaki M, Anindita PD, Ito N, Sugiyama M, Carr M, Fukuhara H, Ose T, Maenaka K, Takada A, Hall WW, Orba Y, Sawa H
The Journal of infectious diseases, 217, 11, 1740, 1749, May 2018, [Peer-reviewed], [International Magazine]
English, Scientific journal, Rabies virus (RABV) is the causative agent of fatal neurological disease. Cellular attachment is the initial and essential step for viral infections. Although extensive studies have demonstrated that RABV uses various target cell molecules to mediate infection, no specific molecule has been identified as an attachment factor for RABV infection. Here we demonstrate that cellular heparan sulfate (HS) supports RABV adhesion and subsequent entry into target cells. Enzymatic removal of HS reduced cellular susceptibility to RABV infection, and heparin, a highly sulfated form of HS, blocked viral adhesion and infection. The direct binding between RABV glycoprotein and heparin was demonstrated, and this interaction was shown to require HS N- and 6-O-sulfation. We also revealed that basic amino acids in the ectodomain of RABV glycoprotein serve as major determinants for the RABV-HS interaction. Collectively, our study highlights a previously undescribed role of HS as an attachment factor for RABV infection.

The C-terminal helix of ribosomal P stalk recognizes a hydrophobic groove of elongation factor 2 in a novel fashion.
Tanzawa T, Kato K, Girodat D, Ose T, Kumakura Y, Wieden HJ, Uchiumi T, Tanaka I, Yao M
Nucleic acids research, 46, 6, 3232, 3244, Oxford University Press (OUP), Apr. 2018, [Peer-reviewed]
Scientific journal

Structural and thermodynamic analyses reveal critical features of glycopeptide recognition by the human PILR alpha immune cell receptor
Atsushi Furukawa, Kosuke Kakita, Tomoki Yamada, Mikihiro Ishizuka, Jiro Sakamoto, Nanao Hatori, Naoyoshi Maeda, Fumina Ohsaka, Takashi Saitoh, Takao Nomura, Kimiko Kuroki, Hisanori Nambu, Hisashi Arase, Shigeki Matsunaga, Masahiro Anada, Toyoyuki Ose, Shunichi Hashimoto, Katsumi Maenaka
JOURNAL OF BIOLOGICAL CHEMISTRY, 292, 51, 21128, 21136, Dec. 2017, [Peer-reviewed]
English, Scientific journal

Rapid Screening by Cell-Based Fusion Assay for Identifying Novel Antivirals of Glycoprotein B-Mediated Herpes Simplex Virus Type 1 Infection
Naoyoshi Maeda, Atsushi Furukawa, Kosuke Kakita, Masahiro Anada, Shunichi Hashimoto, Shigeki Matsunaga, Kimiko Kuroki, Toyoyuki Ose, Akihisa Kato, Jun Arii, Yasushi Kawaguchi, Hisashi Arase, Katsumi Maenaka
BIOLOGICAL & PHARMACEUTICAL BULLETIN, 39, 11, 1897, 1902, Nov. 2016, [Peer-reviewed]
English, Scientific journal

The immunosuppressive effect of domain-deleted dimer of HLA-G2 isoform in collagen-induced arthritis mice
Ami Takahashi, Kimiko Kuroki, Yuki Okabe, Yoshiyuki Kasai, Naoki Matsumoto, Chisato Yamada, Toshiyuki Takai, Toyoyuki Ose, Shigeyuki Kon, Tadashi Matsuda, Katsumi Maenaka
HUMAN IMMUNOLOGY, 77, 9, 754, 759, Sep. 2016, [Peer-reviewed]
English, Scientific journal

Interaction between a Domain of the Negative Regulator of the Ras-ERK Pathway, SPRED1 Protein, and the GTPase-activating Protein-related Domain of Neurofibromin Is Implicated in Legius Syndrome and Neurofibromatosis Type 1
Yasuko Hirata, Hilde Brems, Mayu Suzuki, Mitsuhiro Kanamori, Masahiro Okada, Rimpei Morita, Isabel Llano-Rivas, Toyoyuki Ose, Ludwine Messiaen, Eric Legius, Akihiko Yoshimura
JOURNAL OF BIOLOGICAL CHEMISTRY, 291, 7, 3124, 3134, Feb. 2016, [Peer-reviewed]
English, Scientific journal

Solution structure of an avirulence protein, AVR-Pia, from Magnaporthe oryzae
Toyoyuki Ose, Azusa Oikawa, Yukiko Nakamura, Katsumi Maenaka, Yuya Higuchi, Yuki Satoh, Shiho Fujiwara, Makoto Demura, Teruo Sone, Masakatsu Kamiya
JOURNAL OF BIOMOLECULAR NMR, 63, 2, 229, 235, Oct. 2015, [Peer-reviewed]
English, Scientific journal

Crystal structure of extracellular domain of human lectin-like transcript 1 (LLT1), the ligand for natural killer receptor-P1A
Shunsuke Kita, Haruki Matsubara, Yoshiyuki Kasai, Takaharu Tamaoki, Yuki Okabe, Hideo Fukuhara, Jun Kamishikiryo, Elena Krayukhina, Susumu Uchiyama, Toyoyuki Ose, Kimiko Kuroki, Katsumi Maenaka
EUROPEAN JOURNAL OF IMMUNOLOGY, 45, 6, 1605, 1613, Jun. 2015, [Peer-reviewed]
English, Scientific journal

Roquin binds microRNA-146a and Argonaute2 to regulate microRNA homeostasis
Monika Srivastava, Guowen Duan, Nadia J. Kershaw, Vicki Athanasopoulos, Janet H. C. Yeo, Toyoyuki Ose, Desheng Hu, Simon H. J. Brown, Slobodan Jergic, Hardip R. Patel, Alvin Pratama, Sashika Richards, Anil Verma, E. Yvonne Jones, Vigo Heissmeyer, Thomas Preiss, Nicholas E. Dixon, Mark M. W. Chong, Jeffrey J. Babon, Carola G. Vinuesa
NATURE COMMUNICATIONS, 6, 6253, Feb. 2015, [Peer-reviewed]
English, Scientific journal

Structural Characterization Reveals the Keratinolytic Activity of an Arthrobacter nicotinovorans Protease
Teruo Sone, Yumiko Haraguchi, Aki Kuwahara, Toyoyuki Ose, Megumi Takano, Ayumi Abe, Michiko Tanaka, Isao Tanaka, Kozo Asano
PROTEIN AND PEPTIDE LETTERS, 22, 1, 63, 72, 2015, [Peer-reviewed]
English, Scientific journal

X-ray structures of eIF5B and the eIF5B-eIF1A complex: the conformational flexibility of eIF5B is restricted on the ribosome by interaction with eIF1A
Aiping Zheng, Jian Yu, Reo Yamamoto, Toyoyuki Ose, Isao Tanaka, Min Yao
ACTA CRYSTALLOGRAPHICA SECTION D-BIOLOGICAL CRYSTALLOGRAPHY, 70, Pt 12, 3090, 3098, Dec. 2014, [Peer-reviewed]
English, Scientific journal

Structural basis for simultaneous recognition of an O-glycan and its attached peptide of mucin family by immune receptor PILR alpha
Kimiko Kuroki, Jing Wang, Toyoyuki Ose, Munechika Yamaguchi, Shigekazu Tabata, Nobuo Maita, Seiko Nakamura, Mizuho Kajikawa, Amane Kogure, Takeshi Satoh, Hisashi Arase, Katsumi Maenaka
PROCEEDINGS OF THE NATIONAL ACADEMY OF SCIENCES OF THE UNITED STATES OF AMERICA, 111, 24, 8877, 8882, Jun. 2014, [Peer-reviewed], [Lead author]
English, Scientific journal

Heterologous production, purification, and immunodetection of Magnaporthe oryzae avirulence protein AVR-Pia
Yuki Satoh, Shinsuke Miki, Toyoyuki Ose, Azusa Oikawa, Katsumi Maenaka, Ryouhei Terauchi, Kozo Asano, Teruo Sone
BIOSCIENCE BIOTECHNOLOGY AND BIOCHEMISTRY, 78, 4, 680, 686, Apr. 2014, [Peer-reviewed]
English, Scientific journal

Allosteric Regulation of Epoxide Opening Cascades by a Pair of Epoxide Hydrolases in Monensin Biosynthesis
Atsushi Minami, Toyoyuki Ose, Kyohei Sato, Azusa Oikawa, Kimiko Kuroki, Katsumi Maenaka, Hiroki Oguri, Hideaki Oikawa
ACS CHEMICAL BIOLOGY, 9, 2, 562, 569, Feb. 2014, [Peer-reviewed]
English, Scientific journal

NKG2D Triggers Cytotoxicity in Murine Epidermal gamma delta T Cells via PI3K-Dependent, Syk/ZAP70-Independent Signaling Pathway
Atsuko Ibusuki, Kazuhiro Kawai, Shigeru Yoshida, Youhei Uchida, Ayano Nitahara-Takeuchi, Kimiko Kuroki, Mizuho Kajikawa, Toyoyuki Ose, Katsumi Maenaka, Masanori Kasahara, Takuro Kanekura
JOURNAL OF INVESTIGATIVE DERMATOLOGY, 134, 2, 396, 404, Feb. 2014, [Peer-reviewed]
English, Scientific journal

Crystal Structure of the Lamprey Variable Lymphocyte Receptor C Reveals an Unusual Feature in Its N-Terminal Capping Module
Ryo Kanda, Yoichi Sutoh, Jun Kasamatsu, Katsumi Maenaka, Masanori Kasahara, Toyoyuki Ose
PLOS ONE, 9, 1, e85875, Jan. 2014, [Peer-reviewed]
English, Scientific journal

Erratum: Structure analysis of geranyl pyrophosphate methyltransferase and the proposed reaction mechanism of SAM-dependent C-methylation (Acta Crystallographica Section D: Biological Crystallography (2012) (1558-1569)
Orapin Ariyawutthiphan, Toyoyuki Ose, Atsushi Minami, Sandip Shinde, Muneya Tsuda, Yong-Gui Gao, Min Yao, Hideaki Oikawa, Isao Tanaka
Acta Crystallographica Section D: Biological Crystallography, 69, 12, 2584, Dec. 2013, [Peer-reviewed]
English, Scientific journal

Structural analysis for glycolipid recognition by the C-type lectins Mincle and MCL
Atsushi Furukawa, Jun Kamishikiryo, Daiki Mori, Kenji Toyonaga, Yuki Okabe, Aya Toji, Ryo Kanda, Yasunobu Miyake, Toyoyuki Ose, Sho Yamasaki, Katsumi Maenaka
PROCEEDINGS OF THE NATIONAL ACADEMY OF SCIENCES OF THE UNITED STATES OF AMERICA, 110, 43, 17438, 17443, Oct. 2013, [Peer-reviewed]
English, Scientific journal

Distinct HIV-1 Escape Patterns Selected by Cytotoxic T Cells with Identical Epitope Specificity
Yuichi Yagita, Nozomi Kuse, Kimiko Kuroki, Hiroyuki Gatanaga, Jonathan M. Carlson, Takayuki Chikata, Zabrina L. Brumme, Hayato Murakoshi, Tomohiro Akahoshi, Nico Pfeifer, Simon Mallal, Mina John, Toyoyuki Ose, Haruki Matsubara, Ryo Kanda, Yuko Fukunaga, Kazutaka Honda, Yuka Kawashima, Yasuo Ariumi, Shinichi Oka, Katsumi Maenaka, Masafumi Takiguchi
JOURNAL OF VIROLOGY, 87, 4, 2253, 2263, Feb. 2013, [Peer-reviewed]
English, Scientific journal

Structure analysis of geranyl pyrophosphate methyltransferase and the proposed reaction mechanism of SAM-dependent C-methylation
Orapin Ariyawutthiphan, Toyoyuki Ose, Atsushi Minami, Sandip Sinde, Muneya Tsuda, Yong-Gui Gao, Min Yao, Hideaki Oikawa, Isao Tanaka
ACTA CRYSTALLOGRAPHICA SECTION D-BIOLOGICAL CRYSTALLOGRAPHY, 68, 1558, 1569, Nov. 2012, [Peer-reviewed]
English, Scientific journal

Simultaneous inhibition of Src and Aurora kinases by SU6656 induces therapeutic synergy in human synovial sarcoma growth, invasion and angiogenesis in vivo
Ryuta Arai, Masumi Tsuda, Takuya Watanabe, Toyoyuki Ose, Chikashi Obuse, Katsumi Maenaka, Akio Minami, Yusuke Ohba
EUROPEAN JOURNAL OF CANCER, 48, 15, 2417, 2430, Oct. 2012, [Peer-reviewed]
English, Scientific journal

Crystallization Strategy for the Glycoprotein-Receptor Complex Between Measles Virus Hemagglutinin and Its Cellular Receptor SLAM
Takao Hashiguchi, Toyoyuki Ose, Marie Kubota, Nobuo Maita, Jun Kamishikiryo, Katsumi Maenaka, Yusuke Yanagi
PROTEIN AND PEPTIDE LETTERS, 19, 4, 468, 473, Apr. 2012, [Peer-reviewed]
English, Scientific journal

Crystallization and preliminary X-ray structure analysis of human ribosomal protein L30e
Akiko Kawaguchi, Toyoyuki Ose, Min Yao, Isao Tanaka
ACTA CRYSTALLOGRAPHICA SECTION F-STRUCTURAL BIOLOGY COMMUNICATIONS, 67, 1516, 1518, Dec. 2011, [Peer-reviewed]
English, Scientific journal

Remarkable synergistic effect between MonBI and MonBII on epoxide opening reaction in ionophore polyether monensin biosynthesis
Kyohei Sato, Atsushi Minami, Toyoyuki Ose, Hiroki Oguri, Hideaki Oikawa
TETRAHEDRON LETTERS, 52, 41, 5277, 5280, Oct. 2011, [Peer-reviewed]
English, Scientific journal

Crystallographic Snapshots of Tom20-Mitochondrial Presequence Interactions with Disulfide-Stabilized Peptides
Takashi Saitoh, Mayumi Igura, Yusuke Miyazaki, Toyoyuki Ose, Nobuo Maita, Daisuke Kohda
BIOCHEMISTRY, 50, 24, 5487, 5496, Jun. 2011, [Peer-reviewed]
English, Scientific journal

Crystallization and preliminary X-ray crystallographic study of a methyltransferase involved in 2-methylisoborneol biosynthesis in Streptomyces lasaliensis
Orapin Ariyawutthiphan, Toyoyuki Ose, Muneya Tsuda, YongGui Gao, Min Yao, Atsushi Minami, Hideaki Oikawa, Isao Tanaka
ACTA CRYSTALLOGRAPHICA SECTION F-STRUCTURAL BIOLOGY AND CRYSTALLIZATION COMMUNICATIONS, 67, 417, 420, Mar. 2011, [Peer-reviewed]
English, Scientific journal

Structure of the measles virus hemagglutinin bound to its cellular receptor SLAM
Takao Hashiguchi, Toyoyuki Ose, Marie Kubota, Nobuo Maita, Jun Kamishikiryo, Katsumi Maenaka, Yusuke Yanagi
NATURE STRUCTURAL & MOLECULAR BIOLOGY, 18, 2, 135, U191, Feb. 2011, [Peer-reviewed]
English, Scientific journal

3D1358 The Recognition Mechanism of eIF2β for its partner proteins eIF5 and eIF2β_ε(3D Protein: Structure & Function 3,The 49th Annual Meeting of the Biophysical Society of Japan)
Gai Zuoqi, Kitagawa Yumie, Yao Min, Ose Toyoyuki, Tanaka Yoshikazu, Tanaka Isao
Seibutsu Butsuri, 51, S119, S120, The Biophysical Society of Japan General Incorporated Association, 2011
English

Importance of the Hydrogen Bonding Network Including Asp52 for Catalysis, as Revealed by Asn59 Mutant Hen Egg-white Lysozymes
Toyoyuki Ose, Kimiko Kuroki, Masaaki Matsushima, Katsumi Maenaka, Izumi Kumagai
JOURNAL OF BIOCHEMISTRY, 146, 5, 651, 657, Nov. 2009, [Peer-reviewed]
English, Scientific journal

The structure of alanyl-tRNA synthetase with editing domain
Masaaki Sokabe, Toyoyuki Ose, Akiyoshi Nakamura, Keita Tokunaga, Osamu Nureki, Min Yao, Isao Tanaka
PROCEEDINGS OF THE NATIONAL ACADEMY OF SCIENCES OF THE UNITED STATES OF AMERICA, 106, 27, 11028, 11033, Jul. 2009, [Peer-reviewed]
English, Scientific journal

Tom20 recognizes mitochondrial presequences through dynamic equilibrium among multiple bound states
Takashi Saitoh, Mayumi Igura, Takayuki Obita, Toyoyuki Ose, Rieko Kojima, Katsumi Maenaka, Toshiya Endo, Daisuke Kohda
EMBO JOURNAL, 26, 22, 4777, 4787, Nov. 2007, [Peer-reviewed]
English, Scientific journal

Structural basis of the 3'-end recognition of a leading strand in stalled replication forks by PriA
Kaori Sasaki, Toyoyuki Ose, Naoaki Okamoto, Katsumi Maenaka, Taku Tanaka, Hisao Masai, Mihoko Saito, Tsuyoshi Shirai, Daisuke Kohda
EMBO JOURNAL, 26, 10, 2584, 2593, May 2007, [Peer-reviewed]
English, Scientific journal

Structural basis for dynamic interdomain movement and RNA recognition of the selenocysteine-specific elongation factor SelB
Toyoyuki Ose, Nicolas Soler, Linda Rasubala, Kimiko Kuroki, Daisuke Kohda, Dominique Fourmy, Satoko Yoshizawa, Katsumi Maenaka
STRUCTURE, 15, 5, 577, 586, May 2007, [Peer-reviewed]
English, Scientific journal

Crystal structure of the human monocyte-activating receptor, "Group 2" leukocyte Ig-like receptor A5 (LILRA5/LIR9/ILT11)
Mitsunori Shiroishi, Mizuho Kajikawa, Kimiko Kuroki, Toyoyuki Ose, Daisuke Kohda, Katsumi Maenaka
JOURNAL OF BIOLOGICAL CHEMISTRY, 281, 28, 19536, 19544, Jul. 2006, [Peer-reviewed]
English, Scientific journal

Efficient leukocyte Ig-like receptor signaling and crystal structure of disulfide-linked HLA-G dimer
M Shiroishi, K Kuroki, T Ose, L Rasubala, Shiratori, I, H Arase, K Tsumoto, Kumagai, I, D Kohda, K Maenaka
JOURNAL OF BIOLOGICAL CHEMISTRY, 281, 15, 10439, 10447, Apr. 2006, [Peer-reviewed]
English, Scientific journal

Crystallization and preliminary crystallographic analysis of the N-terminal domain of PriA from Escherichia coli.
Sasaki K, Ose T, Tanaka T, Mizukoshi T, Ishigaki T, Maenaka K, Masai H, Kohda D
Biochimica et biophysica acta, 1764, 157, 160, 1, Jan. 2006, [Peer-reviewed]

Crystallization and preliminary X-ray analysis of mitochondrial presequence receptor Tom20 in complexes with a presequence from aldehyde dehydrogenase
M Igura, T Ose, T Obita, C Sato, K Maenaka, T Endo, D Kohda
ACTA CRYSTALLOGRAPHICA SECTION F-STRUCTURAL BIOLOGY AND CRYSTALLIZATION COMMUNICATIONS, 61, 514, 517, May 2005, [Peer-reviewed]
English, Scientific journal

Crystallization and preliminary X-ray analysis of the mRNA-binding domain of elongation factor SelB in complex with RNA
L Rasubala, D Fourmy, T Ose, D Kohda, K Maenaka, S Yoshizawa
ACTA CRYSTALLOGRAPHICA SECTION F-STRUCTURAL BIOLOGY AND CRYSTALLIZATION COMMUNICATIONS, 61, 296, 298, Mar. 2005, [Peer-reviewed]
English, Scientific journal

Crystallization and preliminary X-ray analysis of alpha-xylosidase from Escherichia coli
M Kitamura, T Ose, M Okuyama, H Watanabe, M Yao, H Mori, A Kimura, Tanaka, I
ACTA CRYSTALLOGRAPHICA SECTION F-STRUCTURAL BIOLOGY AND CRYSTALLIZATION COMMUNICATIONS, 61, 178, 179, Feb. 2005, [Peer-reviewed]
English, Scientific journal

Structural basis for mRNA recognition by elongation factor SelB
S Yoshizawa, L Rasubala, T Ose, D Kohda, D Fourmy, K Maenaka
NATURE STRUCTURAL & MOLECULAR BIOLOGY, 12, 2, 198, 203, Feb. 2005, [Peer-reviewed]
English, Scientific journal

Crystal structure of a biologically functional form of PriB from Escherichia coli reveals a potential single-stranded DNA-binding site
S Shioi, T Ose, K Maenaka, M Shiroishi, Y Abe, D Kohda, T Katayama, T Ueda
BIOCHEMICAL AND BIOPHYSICAL RESEARCH COMMUNICATIONS, 326, 4, 766, 776, Jan. 2005, [Peer-reviewed]
English, Scientific journal

Crystal structure of the ATP-binding cassette of multisugar transporter from Pyrococcus horikoshii OT3
T Ose, T Fujie, M Yao, N Watanabe, Tanaka, I
PROTEINS-STRUCTURE FUNCTION AND BIOINFORMATICS, 57, 3, 635, 638, Nov. 2004, [Peer-reviewed]
English, Scientific journal

Structural and enzymatic properties of 1-aminocyclopropane-1-carboxylate deaminase homologue from Pyrococcus horikoshii
A Fujino, T Ose, M Yao, T Tokiwano, M Honma, N Watanabe, Tanaka, I
JOURNAL OF MOLECULAR BIOLOGY, 341, 4, 999, 1013, Aug. 2004, [Peer-reviewed]
English, Scientific journal

Structure of macrophomate synthase
T Ose, K Watanabe, M Yao, M Honma, H Oikawa, Tanaka, I
ACTA CRYSTALLOGRAPHICA SECTION D-BIOLOGICAL CRYSTALLOGRAPHY, 60, 1187, 1197, Jul. 2004, [Peer-reviewed]
English, Scientific journal

Reaction intermediate structures of 1-aminocyclopropane-1-carboxylate deaminase - Insight into PLP-dependent cyclopropane ring-opening reaction
T Ose, A Fujino, M Yao, N Watanabe, M Honma, Tanaka, I
JOURNAL OF BIOLOGICAL CHEMISTRY, 278, 42, 41069, 41076, Oct. 2003, [Peer-reviewed]
English, Scientific journal

Insight into a natural Diels-Alder reaction from the structure of macrophomate synthase
T Ose, K Watanabe, T Mie, M Honma, H Watanabe, M Yao, H Oikawa, Tanaka, I
NATURE, 422, 6928, 185, 189, Mar. 2003, [Peer-reviewed]
English, Scientific journal

Crystal structure of 1-aminocyclopropane-1-carboxylate deaminase from Hartsenula saturnus
M Yao, T Ose, H Sugimoto, A Horiuchi, A Nakagawa, S Wakatsuki, D Yokoi, T Murakami, M Honma, Tanaka, I
JOURNAL OF BIOLOGICAL CHEMISTRY, 275, 44, 34557, 34565, Nov. 2000, [Peer-reviewed]
English, Scientific journal

Reaction and structure of 1-aminocyclopropane-1-carboxylate deaminase
M Honma, T Murakami, T Ose, H Matsui, Tanaka, I
BIOCHEMISTRY AND MOLECULAR BIOLOGY OF VITAMIN B6 AND PQQ-DEPENDENT PROTEINS, 171, 174, 2000, [Peer-reviewed]
English, International conference proceedings

ミツバチα-アミラーゼの構造と機能
佐分利亘, 高橋有志, 武井梓穂, 尾瀬農之, 森春英, 日本応用糖質科学会大会・応用糖質科学シンポジウム講演要旨集, 73rd-13th, 2024

Molecular basis of inhibition of type II interferon pathway by rabies virus P-protein
南未来, 武川祐一郎, 杉山葵, 飯塚祐太, 稲葉理美, MOSELEY Gregory W., 杉田征彦, 尾瀬農之, 日本ウイルス学会学術集会プログラム・予稿集(Web), 71st, 2024

クライオ電子顕微鏡を用いた,狂犬病ウイルスによるヒトSTAT1経路阻害の構造基盤
南未来, 杉山葵, 武川祐一郎, 姚閔, 姚閔, 稲葉理美, 稲葉理美, 廣瀬未果, 杉田征彦, 尾瀬農之, 尾瀬農之, 日本蛋白質科学会年会(Web), 24th, 2024

狂犬病ウイルスP蛋白質によるSTAT阻害機構解明のための,複合体クライオ電顕構造解析に向けた取り組み
杉山葵, 南未来, 杉田征彦, 廣瀬未果, 喜多俊介, 喜多俊介, 有森貴夫, 高木淳一, 前仲勝実, 前仲勝実, 姚閔, 姚閔, 尾瀬農之, 尾瀬農之, 日本蛋白質科学会年会(Web), 24th, 2024

乳がん特異的キナーゼとアダプター蛋白質が形成する分子集合と酵素活性の相関を検証する
武井梓穂, 宇賀神魁, 中迫純希, 松尾友樹, 神田諒, 川越聡一郎, 齋尾智英, 松田正, 前仲勝実, 姚閔, 尾瀬農之, 日本蛋白質科学会年会(Web), 24th, 2024

X線小角散乱(SEC-SAXS)による,Breast tumor kinaseの活性化機構の解明
宇賀神魁, 武井梓穂, 松田正, 尾瀬農之, 日本蛋白質科学会年会プログラム・要旨集, 23rd (CD-ROM), 2023

アダプター蛋白質介在に伴う,乳がん特異的キナーゼの活性亢進評価
武井梓穂, 宇賀神魁, 中迫純希, 松尾友樹, 神田諒, 前仲勝実, 松田正, 姚閔, 尾瀬農之, 日本蛋白質科学会年会プログラム・要旨集, 23rd (CD-ROM), 2023

新規酵素ガラクトース2-エピメラーゼの諸性質ならびにX線結晶構造の解析
内山昌典, 佐分利亘, 武井梓穂, 尾瀬農之, 森春英, 日本農芸化学会北海道支部学術講演会講演要旨集(Web), 2023, 2023

狂犬病ウイルスPタンパク質によるSTAT阻害機構の解明
南未来, 杉山葵, 武川祐一郎, 姚閔, 姚閔, 廣瀬未果, 杉田征彦, 尾瀬農之, 尾瀬農之, 日本蛋白質科学会年会プログラム・要旨集, 23rd (CD-ROM), 2023

世界初IDP酵素の検証
大和田ゆうき, 澤田光平, 荒井彩花, 田所高志, 南篤志, 前仲勝実, 久米田博之, 姚閔, 及川英秋, 尾瀬農之, 尾瀬農之, 日本結晶学会年会講演要旨集, 2022, 2022

Structure determination of protein-tRNA complex using AlphaFold2 predicted model
碓井拓哉, 大江花, 中村彰良, 尾瀬農之, 尾瀬農之, YAO Min, YAO Min, Photon Factory News, 40, 3, 2022

狂犬病ウイルスP蛋白質による宿主の免疫経路阻害機構の解明
南未来, 杉山葵, 野間井智, JIANG Xinxin, 前仲勝実, YAO Min, 尾瀬農之, 日本蛋白質科学会年会プログラム・要旨集, 21st, 2021

病原性理解のためのLyssavirus P蛋白質C末端ドメイン構造解析
杉山葵, 野間井智, JIANG Xinxin, 南未来, 前仲勝実, 伊藤直人, GOOLEY Paul R., MOSELEY Gregory W., YAO Min, 尾瀬農之, 尾瀬農之, 日本蛋白質科学会年会プログラム・要旨集, 21st, 2021

麻疹ウイルスVタンパク質によるType I IFN経路阻害機構の解明
木本円花, 永野悠馬, 杉山葵, 姚閔, 田所高志, 前仲勝実, 尾瀬農之, 日本結晶学会年会講演要旨集, 2021, 2021

毒ヘビ由来Ca²⁺チャネル阻害型神経毒と毒ヘビ血清蛋白質の複合体構造
田所高志, 岡部由紀, 松原永季, 喜多俊介, 尾瀬農之, 黒木喜美子, 前仲勝実, 寺田成之, 塩井(青木)成留実, 日本結晶学会年会講演要旨集, 2021, 2021

A straightforward approach to antibodies recognising cancer specific glycopeptidic neoepitopes (vol 11, pg 4999, 2020)
Hajime Wakui, Yoshikazu Tanaka, Toyoyuki Ose, Isamu Matsumoto, Koji Kato, Yao Min, Taro Tachibana, Masaharu Sato, Kentaro Naruchi, Fayna Garcia Martin, Hiroshi Hinou, Shin-Ichiro Nishimura, CHEMICAL SCIENCE, 11, 46, 12588, 12589, Dec. 2020
English, Others

Elucidation of PLP binding and substrate recognition mechanism of plant tyrosine decarboxylase
WANG Hang, NAKAGAWA Yusuke, SATOH Yasuharu, YU Jian, OSE Toyoyuki, YAO Min, 量子ビームサイエンスフェスタ(Web), 2019, 2020

リッサウイルスP蛋白質C末端ドメインの構造比較
杉山葵, 野間井智, 蒋欣欣, 南未来, 前仲勝実, 伊藤直人, GOOLEY Paul R., MOSELEY Gregory W., 姚閔, 尾瀬農之, 尾瀬農之, 日本結晶学会年会講演要旨集, 2020 (CD-ROM), 2020

Crystal structure analysis of the C-terminal domain of Duvenhage virus P protein
杉山葵, 蒋欣欣, 前仲勝実, 姚閔, 尾瀬農之, 量子ビームサイエンスフェスタ(Web), 2019, 2020

バクテリアセルロース合成酵素複合体構成成分BosCの構造解析の試み
佐藤亨, 大内香予子, 今井友也, 田島健次, 尾瀬農之, 姚閔, 日本結晶学会年会講演要旨集, 2020 (CD-ROM), 2020

CD1dとアミド基導入抗原複合体のX線結晶構造解析　　　　　　　　　　　　　　　
喜多 俊介, 日下 裕規, 井貫 晋輔, Md. Imran Hossain, 花島 慎弥, 田所 高志, 新山 真由美, 杉山 成, 相羽 俊彦, 尾瀬 農之, 黒木 喜美子, 深瀬 浩一, 村田 道雄, 藤本 ゆかり, 前仲 勝実, 日本結晶学会 講演要旨集, Nov. 2019

Crystal structure analysis of lipid presenting molecule CD1d
喜多俊介, 日下裕規, HOSSAIN Md. Imran, 花島慎弥, 井貫晋輔, 田所高志, 新山真由美, 杉山成, 相羽俊彦, 相羽俊彦, 尾瀬農之, 黒木喜美子, 深瀬浩一, 藤本ゆかり, 村田道雄, 前仲勝実, 量子ビームサイエンスフェスタ(Web), 2018, 2019

イヌジステンパーウイルス受容体結合タンパク質の構造と膜融合の活性化機構
福原秀雄, 酒匂幸, 河村美尋, 梶川瑞穂, 橋口隆生, 竹田誠, PHILIPPE Plattet, 尾瀬農之, 前仲勝実, 日本ウイルス学会学術集会プログラム・予稿集(Web), 67th, 2019

狂犬病ウイルスP蛋白質によるJAK‐STATシグナル阻害機構の解明
杉山葵, 蒋欣欣, 永野悠馬, 野間井智, 若原拓也, 前仲勝実, 姚閔, MOSLEY Gregory, 尾瀬農之, 日本細胞生物学会大会(Web), 71st, ROMBUNNO.2P‐005 (WEB ONLY), 2019
Japanese

IDPを使用した新規ペア型酵素の戦略
澤田光平, 田所高志, 大和田ゆうき, 南篤志, 久米田博之, 斎尾智英, 姚閔, 及川英秋, 前仲勝実, 尾瀬農之, 尾瀬農之, 日本結晶学会年会講演要旨集, 2018, 32, 01 Nov. 2018
Japanese

エポキシド加水分解機能を持つタンパク質の分子進化的考察　　　　　　　　　　　　　　　
飯淵 友直, 尾瀬 農之, 前仲 勝実, 及川 英秋, 園山 正史, 向井 有理, 日本生化学会大会プログラム・講演要旨集, 91回, [2P, 110], Sep. 2018
(公社)日本生化学会, Japanese

脂質抗原提示分子CD1dによる抗原認識機構の解析　　　　　　　　　　　　　　　
日下 裕規, 喜多 俊介, 大野 祐介, 木原 章雄, 尾瀬 農之, 黒木 喜美子, 前仲 勝実, 日本生化学会大会プログラム・講演要旨集, 91回, [3P, 090], Sep. 2018
(公社)日本生化学会, Japanese

エポキシド加水分解機能を持つタンパク質の分子進化的考察　　　　　　　　　　　　　　　
飯淵 友直, 尾瀬 農之, 前仲 勝実, 及川 英秋, 園山 正史, 向井 有理, 日本生化学会大会プログラム・講演要旨集, 91回, [2P, 110], Sep. 2018
(公社)日本生化学会, Japanese

脂質抗原提示分子CD1dによる抗原認識機構の解析　　　　　　　　　　　　　　　
日下 裕規, 喜多 俊介, 大野 祐介, 木原 章雄, 尾瀬 農之, 黒木 喜美子, 前仲 勝実, 日本生化学会大会プログラム・講演要旨集, 91回, [3P, 090], Sep. 2018
(公社)日本生化学会, Japanese

脂質抗原提示分子CD1dによる抗原認識における種差
日下裕規, 喜多俊介, HOSSAIN Imran, 花島慎弥, 井貫晋輔, 新山真由美, 杉山成, 相羽俊彦, 尾瀬農之, 黒木喜美子, 深瀬浩一, 藤本ゆかり, 村田道雄, 前仲勝実, 日本蛋白質科学会年会プログラム・要旨集, 18th, 84, 23 May 2018
Japanese

毒ヘビ由来Ca2+チャネル阻害型神経毒と毒ヘビ血清蛋白質の複合体構造
田所高志, 塩井(青木, 成留実, 岡部由紀, 松原永季, 喜多俊介, 尾瀬農之, 黒木喜美子, 前仲勝実, 寺田成之, 日本蛋白質科学会年会プログラム・要旨集, 18th, 90, 23 May 2018
Japanese

ヒトの自然免疫系を阻害する麻疹ウイルスV蛋白質の機能解析
永野悠馬, 若原拓也, 秦玉瑩, 柳雄介, 前仲勝実, 尾瀬農之, 尾瀬農之, 日本蛋白質科学会年会プログラム・要旨集, 18th, 64, 23 May 2018
Japanese

狂犬病ウイルスP蛋白質によるJAK‐STATシグナル阻害機構の解明
杉山葵, JIANG Xinxin, 永野悠馬, 野間井智, 若原拓也, 前仲勝実, YAO Min, MOSLEY Gregory, 尾瀬農之, 日本蛋白質科学会年会プログラム・要旨集, 18th, 48, 23 May 2018
Japanese

PILRαによる糖ペプチド認識機構の解明　　　　　　　　　　　　　　　
古川 敦, 柿田 浩輔, 荒瀬 尚, 穴田 仁洋, 尾瀬 農之, 橋本 俊一, 前仲 勝実, 日本薬学会年会要旨集, 138年会, 2, 261, 261, Mar. 2018
(公社)日本薬学会, Japanese

PILRαによる糖ペプチド認識機構の解明
古川敦, 柿田浩輔, 荒瀬尚, 穴田仁洋, 尾瀬農之, 橋本俊一, 前仲勝実, 日本薬学会年会要旨集(CD-ROM), 138th, 2, ROMBUNNO.27PA‐am327, 261, Mar. 2018
(公社)日本薬学会, Japanese

Crystal structure and functional analyses of Rabies virus P protein mutants
野間井智, 蒋欣欣, 永野悠馬, 杉山葵, 姚閔, GOOLEY Paul, MOSELEY Gregory, 前仲勝実, 尾瀬農之, 尾瀬農之, 量子ビームサイエンスフェスタ(Web), 2017, 2018

ヒト免疫受容体NKRP1AによるリガンドLLT1の分子認識機構
田所高志, 喜多俊介, 松原永季, 笠井宣征, 玉置貴晴, 岡部由紀, 日下裕規, 石山夢美, 福原秀雄, 上敷領淳, 尾瀬農之, 黒木喜美子, 前仲勝実, 日本生化学会大会(Web), 2017年度, [2P, 0185], Dec. 2017
生命科学系学会合同年次大会運営事務局, Japanese

HIV-2 Nefタンパク質のX線結晶構造　　　　　　　　　　　　　　　
平尾 憲吾, 黒木 喜美子, Andrews Sophie, 尾瀬 農之, Rowland-Jones Sarah, 前仲 勝実, 生命科学系学会合同年次大会, 2017年度, [2P, 0091], Dec. 2017
生命科学系学会合同年次大会運営事務局, English

単純ヘルペスウイルス由来糖蛋白質がヒト受容体を認識するメカニズム
尾瀬農之, 石塚幹広, 古川敦, 黒木喜美子, 前仲勝実, 日本結晶学会年会講演要旨集, 2017, 47, 23 Nov. 2017
Japanese

狂犬病ウイルスの細胞吸着に関与する宿主因子の解析
佐々木道仁, アニンディタ パウリナ, 伊藤直人, 杉山誠, 福原秀雄, 尾瀬農之, 尾瀬農之, 前仲勝実, 澤洋文, 澤洋文, 日本獣医学会学術集会講演要旨集, 160th, 400, 400, 30 Aug. 2017
(公社)日本獣医学会, Japanese

狂犬病ウイルスの細胞吸着に関与する宿主因子の解析　　　　　　　　　　　　　　　
佐々木 道仁, アニンディ・タパウリナ, 伊藤 直人, 杉山 誠, 福原 秀雄, 尾瀬 農之, 前仲 勝実, 澤 洋文, 日本獣医学会学術集会講演要旨集, 160回, 400, 400, Aug. 2017
(公社)日本獣医学会, Japanese

麻疹ウイルスV蛋白質C末端領域の構造的特徴
永野悠馬, 若原拓也, 蒋欣欣, 柳雄介, 前仲勝実, 尾瀬農之, 日本蛋白質科学会年会プログラム・要旨集, 17th, 48, 22 May 2017
Japanese

分泌タンパク質ANGPTL2を介したシグナル伝達を阻害可能な低分子化合物探索
荒牧峻彦, 黒木喜美子, 門松毅, 尾池雄一, 尾瀬農之, 前仲勝実, 日本蛋白質科学会年会プログラム・要旨集, 17th, 147, 22 May 2017
Japanese

トラスツズマブ一本鎖Fv抗体断片の構築および高機能化
田所高志, 中村光太, 坪井晴美, 前田龍, 尾瀬農之, 松田彰, 前仲勝実, 日本蛋白質科学会年会プログラム・要旨集, 17th, 148, 22 May 2017
Japanese

CD1dの一本鎖化による安定化と結晶構造解析
日下裕規, 喜多俊介, HOSSAIN Imuran Md, 花島慎弥, 井貫晋輔, 新山真由美, 杉山成, 尾瀬農之, 黒木喜美子, 藤本ゆかり, 村田道雄, 前仲勝実, 日本蛋白質科学会年会プログラム・要旨集, 17th, 120, 22 May 2017
Japanese

インフルエンザウイルス細胞侵入において鍵となる宿主タンパク質の同定　　　　　　　　　　　　　　　
藤岡 容一朗, 西出 真也, 尾瀬 農之, 加藤 いづみ, 福原 秀雄, 藤岡 真理, 堀内 浩水, 佐藤 絢, Nepal Prabha, 柏木 彩花, Wang Jing, 堀口 美香, Paudel Sarad, 南保 明日香, 宮崎 忠昭, 前仲 勝実, 大場 雄介, 日本細胞生物学会大会講演要旨集, 69回, 61, 61, May 2017
(一社)日本細胞生物学会, Japanese

インフルエンザウイルス細胞侵入において鍵となる宿主タンパク質の同定
藤岡容一朗, 西出真也, 尾瀬農之, 加藤いづみ, 福原秀雄, 藤岡真理, 堀内浩水, 佐藤絢, NEPAL Prabha, 柏木彩花, WANG Jing, 堀口美香, PAUDEL Sarad, 南保明日香, 宮崎忠昭, 前仲勝実, 前仲勝実, 大場雄介, 日本細胞生物学会大会(Web), 69th, ROMBUNNO.T8‐04(P2‐028) (WEB ONLY), 61, May 2017
(一社)日本細胞生物学会, Japanese

Insight into the activation mechanism of Brk
松尾友樹, 神田諒, 西條慎也, 清水伸隆, 前仲勝実, 尾瀬農之, 量子ビームサイエンスフェスタ(Web), 2016, 2017

自然免疫受容体Mincleの糖脂質認識機構
古川敦, 須知佑介, 池野里紗, 松丸尊紀, 上敷領淳, 齊藤貴士, 尾瀬農之, 山崎晶, 前仲勝実, 日本薬学会年会要旨集(CD-ROM), 137th, ROMBUNNO.S12‐1, 2017
Japanese

アンジオポエチン様因子Angptl2の結晶構造解析および免疫受容体LILRB2との相互作用解析
荒牧峻彦, 黒木喜美子, 門松毅, 寺田和豊, 尾池雄一, 尾瀬農之, 前仲勝実, 日本蛋白質科学会年会プログラム・要旨集, 16th, 104, 19 May 2016
Japanese

I型IFNシグナルを回避する麻疹ウイルスVタンパク質の分子基盤解析
永野悠馬, 若原拓也, 野間井智, 市川聡, 柳雄介, 前仲勝実, 尾瀬農之, 日本蛋白質科学会年会プログラム・要旨集, 16th, 85, 19 May 2016
Japanese

ヘパラン硫酸を介した狂犬病ウイルスの細胞吸着機構の解析
佐々木道仁, アニンディタ パウリナ, 伊藤直人, 杉山誠, 福原秀雄, 尾瀬農之, 前仲勝実, 大場靖子, 澤洋文, 澤洋文, 日本分子生物学会年会プログラム・要旨集(Web), 39th, ROMBUNNO.2P‐0314 (WEB ONLY), 2016
Japanese

狂犬病ウイルスGタンパク質特異的抗体の中和機構の解明
有坂知朗, 荒牧峻彦, 青木亨丞, 伊藤直人, 杉山誠, 尾瀬農之, 福原秀雄, 前仲勝実, 日本分子生物学会年会プログラム・要旨集(Web), 39th, ROMBUNNO.1P‐0598 (WEB ONLY), 2016
Japanese

構造分子生物学・生化学の進展 糖脂質認識C型レクチン受容体Mincleの構造解析と新規アジュバント探索　　　　　　　　　　　　　　　
古川 敦, 上敷領 淳, 須知 裕介, 池野 里紗, 松丸 尊紀, 児玉 耕太, 尾瀬 農之, 山崎 晶, 前仲 勝実, 日本生化学会大会・日本分子生物学会年会合同大会講演要旨集, 88回・38回, [3W7, 6], Dec. 2015
(公社)日本生化学会, English

Lectin‐like transcript1(LLT1)C型レクチン様ドメインの結晶構造解析
喜多俊介, 田所高志, 松原永季, 笠井宣征, 玉置貴晴, 岡部由紀, 日下裕規, 石山夢美, 福原秀雄, 上敷領淳, KRAYUKHINA Elena, KRAYUKHINA Elena, 内山進, 尾瀬農之, 黒木喜美子, 前仲勝実, 日本生化学会大会(Web), 88回・38回, [3T17p, 02(3P0328)], Dec. 2015
(公社)日本生化学会, Japanese

構造生物学が推進する医薬基盤とその応用 抑制型免疫受容体PILRαによるヘルペスウイルス1型由来O型糖鎖およびペプチド配列の同時認識と創薬
尾瀬農之, 古川敦, 黒木喜美子, 前仲勝実, 医学のあゆみ, 254, 8, 559, 565, Aug. 2015
医歯薬出版(株), Japanese

ポリエーテル化合物生合成酵素の結晶構造解析
尾瀬農之, 笈川あずさ, 野間井智, 澤田光平, 道仙卓也, 南篤志, 及川英秋, 前仲勝実, 日本蛋白質科学会年会プログラム・要旨集, 15th, 67, 26 May 2015
Japanese

単純ヘルペスウイルス侵入阻害糖ペプチドの最適化を目指した熱力学的相互作用解析およびX線結晶構造解析
山田友樹, 古川敦, 柿田浩輔, 坂本二郎, 前田直良, 逢坂文那, 斉藤貴士, 黒木喜美子, 荒瀬尚, 穴田仁洋, 尾瀬農之, 橋本俊一, 前仲勝実, 日本蛋白質科学会年会プログラム・要旨集, 15th, 90, 26 May 2015
Japanese

JAK‐STAT経路を阻害するウイルスタンパク質の機能・構造解析
永野悠馬, 野間井智, 前仲勝実, 柳雄介, 尾瀬農之, 日本蛋白質科学会年会プログラム・要旨集, 15th, 88, 26 May 2015
Japanese

アンジオポエチン様因子Angptl2の発現系構築と免疫受容体LILRB2との相互作用解析
荒牧峻彦, 黒木喜美子, 門松毅, 寺田和豊, 尾池雄一, 尾瀬農之, 前仲勝実, 日本蛋白質科学会年会プログラム・要旨集, 15th, 138, 26 May 2015
Japanese

単純ヘルペスウイルスがペア型レセプターPILRαを利用するメカニズム
尾瀬農之, 黒木喜美子, 山口宗親, 田畑栄一, 真板宣夫, 梶川瑞穂, 中村聖子, WANG Jing, 佐藤毅史, 荒瀬尚, 前仲勝実, 物構研サイエンスフェスタ要旨集, 3rd, 79, 2015
Japanese

Mechanisms of lyssavirus evasion of innate immunity and roles in lethal rabies disease
Linda Wiltzer, Aaron Brice, Kim Lieu, Angela Harrison, Sibil Oksayan, Toby Bell, Donna Whelan, Alamgir Hossain, Patrick Shillling, Kazuma Okada, Satoko Yamaoka, Florence Larrous, Herve Bourhy, Toyoyuki Ose, Paul Gooley, David A. Jans, Danielle Blondel, Naoto Ito, Gregory Moseley, CYTOKINE, 70, 1, 60, 60, Nov. 2014
English, Summary international conference

単純ヘルペスウイルス1型感染阻害物質の探索
前田直良, 古川敦, 坂本二郎, 黒木喜美子, 尾瀬農之, 柿田浩輔, 穴田仁洋, 橋本俊一, 有井潤, 加藤哲久, 川口寧, 荒瀬尚, 前仲勝実, 日本ウイルス学会学術集会プログラム・抄録集, 62nd, 273, 31 Oct. 2014
Japanese

Structural Analysis for the Interaction of sialic T antigen glycopeptide of HSV-1 with Entry Receptor PILR alpha
Takao Nomura, Jiro Sakamoto, Fumina Oosaka, Kosuke Kakita, Atsushi Furukawa, Masahiro Anada, Shunichi Hashimoto, Kimiko Kuroki, Toyoyuki Ose, Hisashi Arase, Takashi Saitoh, Katsumi Maenaka, PROTEIN SCIENCE, 23, 230, 230, Jul. 2014
English, Summary international conference

C型レクチン様受容体MincleとMCLの糖脂質認識機構
古川敦, 上敷領淳, 森大輝, 豊永憲司, 岡部由紀, 藤司亜也, 神田諒, 三宅靖延, 尾瀬農之, 山崎晶, 前仲勝実, 日本蛋白質科学会年会プログラム・要旨集, 14th, 121, 26 May 2014
Japanese

2SBP-02 Structural basis for glycolipid recognition mechanism by C-type lectin like receptor, Mincle(New Development of Structural Cell Biology in Signal Transduction,Symposium,The 52th Annual Meeting of the Biophysical Society of Japan(BSJ2014))
Furukawa Atsushi, Kamishikiryo Jun, Ose Toyoyuki, Yamasaki Sho, Maenaka Katsumi, Seibutsu Butsuri, 54, 1, S131, 2014
一般社団法人 日本生物物理学会, English

単純ヘルペスウイルスがペア型レセプターPILRαを利用する分子基盤
尾瀬農之, 黒木喜美子, 山口宗親, 田畑栄一, 真板宣夫, 梶川瑞穂, 中村聖子, WANG Jing, 佐藤毅史, 荒瀬尚, 前仲勝実, 日本結晶学会年会講演要旨集, 2014, 110, 2014
Japanese

Structure analysis of geranyl pyrophosphate methyltransferase and the proposed reaction mechanism of SAM-dependent C-methylation (vol D68, pg 1558, 2012)
Orapin Ariyawutthiphan, Toyoyuki Ose, Atsushi Minami, Sandip Shinde, Muneya Tsuda, Yong-Gui Gao, Min Yao, Hideaki Oikawa, Isao Tanaka, ACTA CRYSTALLOGRAPHICA SECTION D-BIOLOGICAL CRYSTALLOGRAPHY, 69, 2584, 2584, Dec. 2013
English, Others

monensin生合成酵素の結晶構造解析
道仙卓也, 南篤志, 笈川あずさ, 佐藤恭平, 前仲勝実, 及川英秋, 尾瀬農之, 日本結晶学会年会講演要旨集, 2013, 75, 12 Oct. 2013
Japanese

MurDの大量発現系の構築と結晶化
笈川あずさ, 北辻千展, 尾瀬農之, 前仲勝実, 日本蛋白質科学会年会プログラム・要旨集, 13th, 123, 31 May 2013
Japanese

C型レクチン様受容体Mindeの糖脂質認識機構
古川敦, 上敷領淳, 岡部由紀, 神田涼, 豊永憲司, 森大輝, 三宅靖延, 山崎晶, 尾瀬農之, 前仲勝実, 日本蛋白質科学会年会プログラム・要旨集, 13th, 115, 31 May 2013
Japanese

モネンシン生合成酵素の結晶構造解析
尾瀬農之, 笈川あずさ, 南篤志, 佐藤恭平, 及川英秋, 前仲勝実, 日本蛋白質科学会年会プログラム・要旨集, 13th, 122, 31 May 2013
Japanese

樹状細胞免疫受容体DCIRの発現・精製およびX線結晶構造解析
福原秀雄, 松原永季, 神田諒, 矢部力朗, 海部知則, 尾瀬農之, 岩倉洋一郎, 前仲勝実, 日本蛋白質科学会年会プログラム・要旨集, 13th, 119, 31 May 2013
Japanese

パラミクソウイルス侵入機構と阻害剤開発
東端将哲, 福原秀雄, 齊藤貴士, 陳甦ぜい, 逢坂文那, 児玉耕太, 尾瀬農之, 前仲勝実, 日本薬学会年会要旨集(CD-ROM), 133年会, 1, 100, 100, Mar. 2013
(公社)日本薬学会, Japanese

結晶構造解析に向けた,乳がん特異的キナーゼBrkとアダプタータンパク質STAP‐2の機能解析
神田諒, 関根勇一, 前仲勝実, 松田正, 尾瀬農之, 日本分子生物学会年会プログラム・要旨集(Web), 36th, 2P-0302 (WEB ONLY), 2013
Japanese

麻疹ウイルス及び犬ジステンパーウイルスH蛋白質の構造学的研究
尾瀬農之, 橋口隆生, 酒匂幸, 伊藤由梨, 福原秀雄, 黒木喜美子, 柳雄介, 竹田誠, 前仲勝実, 日本蛋白質科学会年会プログラム・要旨集, 12th, 83, 31 May 2012
Japanese

機能および構造解析に向けた細胞表面蛋白質の調製法
福原秀雄, 橋口隆生, 黒木喜美子, 尾瀬農之, 前仲勝実, 日本蛋白質科学会年会プログラム・要旨集, 12th, 19, 31 May 2012
Japanese

ヤツメウナギ由来可変性リンパ球受容体VLRCの結晶構造
神田諒, 須藤洋一, 笠松純, 笠原正典, 前仲勝実, 尾瀬農之, 日本蛋白質科学会年会プログラム・要旨集, 12th, 117, 31 May 2012
Japanese

ロバスト設計最適化支援ツールを用いたin silicoスクリーニング
児玉耕太, 山口政隆, 小川修一, 尾瀬農之, 前仲勝実, 日本薬学会年会要旨集, 132nd, 4, 115, 05 Mar. 2012
Japanese

ヒト免疫制御受容体LILRB1とリガンドHLA‐Gの複合体の結晶構造解析
黒木喜美子, 松原永季, 神田諒, 上敷領淳, 尾瀬農之, 福永裕子, 前仲勝実, 日本分子生物学会年会プログラム・要旨集(Web), 35th, 1P-0060 (WEB ONLY), 2012
Japanese

樹状細胞免疫受容体DCIRの結晶化と構造解析
福原秀雄, 松原永季, 神田諒, 矢部力朗, 海部知則, 上敷領淳, 尾瀬農之, 岩倉洋一郎, 前仲勝実, 日本分子生物学会年会プログラム・要旨集(Web), 35th, 1P-0056 (WEB ONLY), 2012
Japanese

SrcおよびAuroraキナーゼ2重阻害による滑膜肉腫の相乗的in vivo抗腫瘍効果(Dual inhibition of Src and Aurora kinases abrogates tumor growth, invasion, and angiogenesis of synovial sarcoma in vivo)　　　　　　　　　　　　　　　
津田 真寿美, 新井 隆太, 渡部 琢哉, 尾瀬 農之, 小布施 力史, 前仲 勝実, 三浪 明男, 大場 雄介, 日本癌学会総会記事, 70回, 274, 274, Sep. 2011
日本癌学会, English

多様な形態を有するHLA‐G蛋白質とヒト免疫制御受容体との分子認識
黒木喜美子, 福永裕子, 上敷領淳, 白石充典, 尾瀬農之, 前仲勝実, 日本薬学会年会要旨集, 131年会, 1, 188, 188, Mar. 2011
(公社)日本薬学会, Japanese

イネいもち病菌の非病原性タンパク質AVR-Piaの分子間相互作用の解析
佐藤佑樹, 三木慎介, 尾瀬農之, 奥山雄大, 神崎洋之, 寺内良平, 曾根輝雄, 日本植物病理学会大会プログラム・講演要旨予稿集, 2011, 2011

SU6656 suppresses human synovial sarcoma progression through Src and Aurora kinase inhibition
Arai Ryuta, Tsuda Masumi, Watanabe Takuya, Ose Toyoyuki, Obuse Chikashi, Maenaka Katsumi, Minami Akio, Ohba Yusuke, 日本分子生物学会年会プログラム・要旨集(Web), 34th, WEB ONLY 2P-0690, 2011
Summary national conference

ヤツメウナギ由来可変性リンパ球受容体の結晶学的研究
神田諒, 須藤洋一, 笠松純, 笠原正典, 前仲勝実, 尾瀬農之, 日本結晶学会年会講演要旨集, 2011, 90, 2011
Japanese

麻疹ウイルスH蛋白質と受容体の複合体構造
尾瀬農之, 橋口隆生, 久保田万理恵, 真板宣夫, 柳雄介, 前仲勝実, 日本結晶学会年会講演要旨集, 2011, 87, 2011
Japanese

CDV‐Hの結晶構造解析と機能解析
伊藤由梨, 福原秀雄, 酒匂幸, 橋口隆生, 梶川瑞穂, 竹田誠, 柳雄介, 尾瀬農之, 前仲勝実, 日本結晶学会年会講演要旨集, 2011, 90, 2011
Japanese

ヒト免疫制御受容体LILRB1とリガンドHLA‐Gの複合体の結晶構造解析
松原永季, 黒木喜美子, 上敷領淳, 尾瀬農之, 前仲勝実, 日本結晶学会年会講演要旨集, 2011, 95, 2011
Japanese

HIV由来変異ペプチドによるKIR2DL1を介した免疫逃避機構の分子基盤
黒木喜美子, 上敷領淳, 尾瀬農之, THANANCHAI Hathairat, MAKADZANGE Tariro, ROWLAND‐JONES Sarah, DONG Tao, 前仲勝実, 日本結晶学会年会講演要旨集, 2011, 94, 2011
Japanese

HSV‐1のエンベロープ蛋白質gBペプチドおよびペア型レセプターPILRαの複合体結晶構造解析
尾瀬農之, 山口宗親, WAN Jing, 黒木喜美子, 田畑栄一, 真板宣夫, 中村聖子, 梶川瑞穂, 白鳥行大, 佐藤毅史, 荒瀬尚, 前仲勝実, 日本ウイルス学会学術集会プログラム・抄録集, 58th, 420, 15 Oct. 2010
Japanese

クラスI認識受容体の構造とリガンド認識
黒木喜美子, 福永裕子, 上敷領淳, 白石充典, 尾瀬農之, 前仲勝実, MHC, 17, 2, 136, 136, Aug. 2010
日本組織適合性学会, Japanese

ヒトのセレノシステイン導入系におけるL30の役割
川口亜希子, 尾瀬農之, 中村彰良, 喜多俊介, 姚閔, 田中勲, 日本蛋白質科学会年会プログラム・要旨集, 10th, 110, 15 May 2010
Japanese

大腸菌23S rRNAメチル基転移酵素YcbYのRNA結合能
喜多俊介, 木村聡, 尾瀬農之, 鈴木勉, 姚閔, 田中勲, 日本蛋白質科学会年会プログラム・要旨集, 10th, 128, 15 May 2010
Japanese

麻疹ウイルスによる細胞侵入機構の構造基盤
橋口隆生, 尾瀬農之, 上敷領淳, 柳雄介, 前仲勝実, 日本蛋白質科学会年会プログラム・要旨集, 10th, 32, 15 May 2010
Japanese

細胞表面において生体防御に関わる分子認識の構造生物学
福原秀雄, 橋口隆生, 黒木喜美子, 白石充典, 佐々木香織, 梶川瑞穂, 尾瀬農之, 前仲勝実, 日本蛋白質科学会年会プログラム・要旨集, 10th, 32, 15 May 2010
Japanese

酢酸菌由来CesDとCcpAxの相互作用解析
藤原孝彰, 尾瀬農之, 姚閔, 田島健次, 田中勲, 日本蛋白質科学会年会プログラム・要旨集, 10th, 144, 15 May 2010
Japanese

ペア型レセプターPILRα及びHSV‐1のエンベロープ蛋白質gB模倣ペプチド複合体結晶構造
尾瀬農之, 山口宗親, 黒木喜美子, 田畑栄一, 真板宣夫, 梶川瑞穂, 中村聖子, WANG Jing, 佐藤毅史, 荒瀬尚, 前仲勝実, 日本結晶学会年会講演要旨集, 2010, 97, 2010
Japanese

麻疹ウイルスHタンパク質と受容体SLAMの複合体の結晶構造と麻疹ウイルスの細胞侵入機構
橋口隆生, 尾瀬農之, 上敷領淳, 竹田誠, 前仲勝実, 柳雄介, 日本ウイルス学会学術集会プログラム・抄録集, 57th, 149, 01 Oct. 2009
Japanese

Paired Immunoglobulin(Ig)Like type 2 Receptor(PILR)αによるGlycoprotein B(gB)認識機構の解明
山口宗親, 黒木喜美子, 田畑栄一, 真板宣夫, 梶川瑞穂, 尾瀬農之, 中村聖子, 王静, 佐藤毅, 荒瀬尚, 前仲勝実, 日本ウイルス学会学術集会プログラム・抄録集, 57th, 149, 01 Oct. 2009
Japanese

真正細菌におけるセレノシステイン特異的翻訳伸長因子SelBとSECIS mRNAの相互作用基盤
尾瀬農之, RASUBALA Linda, 神田大輔, SOLER Nicolas, 吉澤聡子, FOURMY Dominique, 前仲勝実, 生化学, ROMBUNNO.2S8A-2, 25 Sep. 2009
Japanese

酢酸菌由来セルロース合成関連タンパク質の調製
藤原孝彰, 尾瀬農之, 田島健次, 田中勲, YAO Min, セルロース学会年次大会講演要旨集, 16th, 151, 152, 20 Jun. 2009
Japanese

Pyrococcus horikoshii由来アラニルtRNA合成酵素の結晶構造
徳永啓太, 曽我部正彰, 尾瀬農之, 中村彰良, 濡木理, 姚閔, 田中勲, PFシンポジウム要旨集, 26th, 2009

アラニルtRNA合成酵素の結晶構造より提唱されたtRNA再結合モデル
徳永啓太, 曽我部正彰, 尾瀬農之, 中村彰良, 濡木理, 姚閔, 田中勲, 日本蛋白質科学会年会プログラム・要旨集, 9th, 2009

マウスMHCクラスIb分子MILL2の結晶構造解析
梶川瑞穂, 尾瀬農之, 笠原正典, 前仲勝実, 生化学, 81回・31回, 2T20-1, 1, Nov. 2008
(公社)日本生化学会, Japanese

“21番目のアミノ酸”セレノシステインを取り込むための分子基盤
尾瀬農之, RASUBALA Linda, 神田大輔, SOLER Nicolas, 吉澤聡子, FOURMY Dominique, 前仲勝実, 補体シンポジウム講演集, 45th, 194, 194, Jul. 2008
(一社)日本補体学会, Japanese

複数の結合様式を介したミトコンドリアTom20によるプレ配列の認識機構
齊藤貴士, 井倉真由美, 尾瀬農之, 前仲勝実, 神田大輔, 日本蛋白質科学会年会プログラム・要旨集, 8th, 19, 23 May 2008
Japanese

構造基盤解析に向けたミトコンドリアTom20‐プレ配列ペプチド複合体の安定化
齊藤貴士, 井倉真由美, 尾瀬農之, 帯田孝之, 小島理恵子, 前仲勝実, 神田大輔, Abstr Annu Meet NMR Soc Jpn, 46th, 164, 165, 11 Sep. 2007
Japanese

PriAタンパク質によるDNA3′末端の塩基非選択的認識機構の解明
佐々木香織, 尾瀬農之, 岡本直明, 田中卓, 前仲勝実, 正井久雄, 斉藤美保子, 白井剛, 神田大輔, 日本蛋白質科学会年会プログラム・要旨集, 7th, 50, 07 May 2007
Japanese

ミトコンドリアTom20によるプレ配列の動的認識機構:緩和解析からのアプローチ
齊藤貴士, 井倉真由美, 尾瀬農之, 帯田孝之, 小島理恵子, 前仲勝実, 神田大輔, 日本蛋白質科学会年会プログラム・要旨集, 7th, 38, 07 May 2007
Japanese

Molecular basis for LILR recognition of HLA-G and its dimer
M. Shiroishi, K. Kuroki, T. Ose, L. Rasubala, I. Shiratori, H. Arase, D. Kohda, K. Maenaka, TISSUE ANTIGENS, 68, 4, CP6, CP6, Oct. 2006
English, Summary international conference

Molecular Basis for the Selenocysteine Incorporation
OSE Toyoyuki, RASUBALA Linda, YOSHIZAWA Satoko, FOURMY Dominique, KOHDA Daisuke, MAENAKA Katsumi, Biophysics, 46, 2, 102, 105, 25 Mar. 2006
日本生物物理学会, Japanese

Crystallization and preliminary crystallographic analysis of the N-terminal domain of PriA from Escherichia coli
K Sasaki, T Ose, T Tanaka, T Mizukoshi, T Ishigaki, K Maenaka, H Masai, D Kohda, BIOCHIMICA ET BIOPHYSICA ACTA-PROTEINS AND PROTEOMICS, 1764, 1, 157, 160, Jan. 2006
English

ミトコンドリアプレ配列認識のためのTom20タンパク質の動的構造
井倉真由美, 帯田孝之, 尾瀬農之, 遠藤斗志也, 前仲勝実, 神田大輔, 日本分子生物学会年会講演要旨集, 28th, 602, 25 Nov. 2005
Japanese

E.coli由来PriAのN末端ドメインによるDNA3′末端認識の構造的基盤
佐々木香織, 尾瀬農之, 田中卓, 岡本直明, 前仲勝実, 正井久雄, 神田大輔, 日本分子生物学会年会講演要旨集, 28th, 393, 25 Nov. 2005
Japanese

21番目のアミノ酸セレノシステインを取り込むための分子基盤
尾瀬 農之, Rasubala Linda, 神田 大輔, 構造生物, 11, 1, 21, 30, 2005
国際科学振興財団構造生物学坂部プロジェクト, Japanese

大腸菌由来PriAとオリゴヌクレオチドとの複合体の結晶化
佐々木香織, 尾瀬農之, 岡本直明, 田中卓, 正井久雄, 前仲勝実, 神田大輔, 日本分子生物学会年会プログラム・講演要旨集, 27th, 618, 25 Nov. 2004
Japanese

ラットTom20タンパク質とALDHプレ配列複合体のX線結晶構造解析
井倉真由美, 帯田孝之, 尾瀬農之, 遠藤斗志也, 前仲勝実, 神田大輔, 日本分子生物学会年会プログラム・講演要旨集, 27th, 732, 25 Nov. 2004
Japanese

ミトコンドリア・プレ配列受容体Tom20‐ALDHプレ配列複合体の結晶構造と緩和解析
帯田孝之, 井倉真由美, 尾瀬農之, 前仲勝実, 遠藤斗志也, 神田大輔, NMR討論会講演要旨集, 43rd, 212, 213, 10 Nov. 2004
Japanese

大腸菌yicl産物の機能およびX線構造解析
奥山正幸, 尾瀬農之, 北村百世, 森春英, 千葉誠哉, 木村淳夫, 田中勲, J Appl Glycosci, 51, Suppl., 40, 20 Jul. 2004
Japanese

Crystal Structure Analysis of Natural Diels-Alderase
OSE Toyoyuki, YAO Min, OIKAWA Hideaki, TANAKA Isao, Nihon Kessho Gakkaishi, 45, 6, 384, 390, 25 Dec. 2003
The crystal structure of natural Diels-Alderase, macrophomate synthase, in complex with pyruvate was determined at 1.7 Å resolution. The structure explains broad substrate specificity and provides mechanistic information regarding the multiple catalyst of this enzyme. The active site reveals several ingenious features for catalyzing the Diels-Alder reaction, which include the existence of residues for making catalytically important hydrogen bonds to the 2-pyrone. The rather large and hydrophobic milieu in the active site cavity allows a large-scale structural reorganization of the product i..., The Crystallographic Society of Japan, Japanese

41 Intermolecular Diels-Alderase, Macrophomate Synthase : Proposal for Reaction Mechanism Based on the Structure of the Active Site
Oikawa Hideaki, Watanabe Kenji, Mie Takashi, Ichihara Akitami, Honma Mamoru, Ose Toyoyuki, Yao Min, Tanaka Isao, Symposium on the Chemistry of Natural Products, symposium papers, 44, 241, 246, 01 Sep. 2002
Macrophomate synthase catalyzes unusual multistep reaction from oxalacetate and 2-pyrone to fungal metabolite, macrophomate involving Diels-Alder reaction. To elucidate its reaction mechanism, requirement of metal ion, kinetic parameter and stereochemical course were investigated. In addition, the structure of the active site of macrophomate synthase was determined by X-ray crystallography., Symposium on the chemistry of natural products, Japanese

植物ホルモンの生合成を阻害する蛋白質の立体構造解析
尾瀬 農之, 姚 閔, 田中 勲, 杉山産業化学研究所年報, 1999, 73, 87, 1999
杉山産業化学研究所, Japanese

量子生命科学ハンドブック
尾瀬農之,小林正人, 第3節 水素原子核の量子効果がもたらす特徴的な生体内化学反応とその量子化学計算技術
エヌ・ティー・エス, Mar. 2024, 9784860438821, iii, x, 346, viiip, Japanese, [Contributor]

出力多様性を司るSTAT多量体化の構造基盤と，それに抗するウイルス蛋白質の解析
科学研究費助成事業
01 Apr. 2024 - 31 Mar. 2028
尾瀬 農之, 久米田 博之, 杉田 征彦, 姚 閔
日本学術振興会, 基盤研究(B), 北海道大学, 24K01959

Studies on molecular mechanism of effector-mediated establishment of host specificity in plant pathogens and their application
Grants-in-Aid for Scientific Research
05 Jul. 2021 - 31 Mar. 2026
高野 義孝, 大木 進野, 尾瀬 農之
本研究では、宿主特異性成立に関与する植物病原菌エフェクター（EPC1、EPC2、EPC3）の解析を起点に、エフェクターを介した植物病原菌の宿主特異性成立の分子基盤を解明することを目的とする。本年度は、まずエフェクターEPC1、EPC2、EPC3をベンサミアナタバコ葉に一過的に発現させ、その発現が同植物のPAMP誘導免疫に与える影響を調べた。その結果、EPC1、EPC2、EPC3いずれも、細菌のPAMP（flg22）が引き起こす活性酸素生成を抑制することを明らかにした。続いて、エピトープタグを付加した各EPCエフェクターをベンサミアナタバコ葉およびメロン子葉に一過的に発現させ、続いて免疫沈降解析を実施し、共沈降してくるタンパク質について、LC-MS/MS解析を実施し、標的因子候補のリスト化を完了した。また、EPC3の特定の領域について、その領域のみで機能が保持されることを明らかにし、さらにその領域を発現させた場合は、発現タンパク質が可溶化することを見出している。ウリ科作物への宿主特異性に関わるエフェクターを網羅的に同定するために、ウリ類炭疽病菌とウリ類炭疽病菌が属するオービクラクレード内の別種に対する比較オミクス解析を実施し、ウリ類炭疽病菌の分離株すべてにおいて存在し、他種では必ずしも存在しない遺伝子群、あるいは、ウリ類炭疽病菌において発現するが他種では必ずしも発現しない遺伝子群の選抜を完了した。さらに最近になり、EPC4と命名した新規のエフェクターの発見に成功している。EPC4は、EPC1、EPC2、EPC3と同様にウリ科作物への病原性には必要である一方、ベンサミアナタバコへの病原性には必須ではない。EPC4について、その一過的発現解析を実施した結果、EPC4がflg22が誘導する活性酸素生成を抑制することを見出している。
Japan Society for the Promotion of Science, Grant-in-Aid for Scientific Research (S), Kyoto University, 21H05032

世界初IDP型酵素の存在意義と構造形成機構の解明
科学研究費助成事業
30 Jun. 2022 - 31 Mar. 2024
久米田 博之, 姚 閔, 尾瀬 農之
立体構造を形成しないIDPとして特に研究が進んでいる蛋白質は，高等生物シグナル伝達系や転写翻訳系に多く含まれ，他の蛋白質との相互作用時に過渡的部分構造形成をおこなうものも多い。一方で，ポリペプチド鎖の大部分がIDPで構成される酵素はこれまで報告が無い。天然に存在するポリエーテル化合物は，多様かつ強力な生物活性を示し，産業応用や細胞生物学への利用が多く見られる。これらのポリエーテル化合物は、テトラヒドロフラン環やテトラヒドロピラン環が多くの不斉点を介し連結されており，構造多様性はエーテル環の数や順序，大きさが決定している。ポリエーテル骨格の構築機構は，鎖状ポリオレフィン前駆体がエポキシ化され，引き続き位置選択的なエポキシド開環反応によりポリエーテル骨格が連鎖的に構築される。私達は，monensin生合成をモデルケースとしてポリエーテル骨格構築機構の解明に取り組んできた。Monensinの場合，その骨格を構築するために3 回の5-exo 酵素環化反応が必要である。これまでポリエーテル天然物生合成経路に存在するエポキシド開環エーテル環化酵素はIDP型の酵素であることを提唱し，構造形成機構や存在意義の解析を進めてきた。結晶構造解析，1H-15N HSQC (NMR), CD, DSC, Native-MSを組み合せ，モネンシン生合成経路中に存在する蛋白質のうち，１つは酵素活性を持たず，もう一方の構造形成を補助するシャペロン様分子であることを示すことができた。一般的にポリエーテル環化反応進行のためには，長大な基質を収納するポケットが必要であり，また，環化反応をうける基質のエポキシドおよび水酸基周辺の構造変化に対応できる柔軟な活性部位が必要である。我々のペア型酵素仮説を使用すると，酵素がこういった課題を解決するために採用している戦略を説明できる。
日本学術振興会, 挑戦的研究(萌芽), 北海道大学, 22K19282

Molecular basis of counteraction against the JAK-STAT signaling pathway by virus
Grants-in-Aid for Scientific Research
01 Apr. 2021 - 31 Mar. 2024
尾瀬 農之, 久米田 博之, 杉田 征彦, 于 健
Japan Society for the Promotion of Science, Grant-in-Aid for Scientific Research (B), Hokkaido University, 21H02408

新規に開発したＢＬＬナノ構造体による、タンパク質結晶核形成過程の研究
科学研究費助成事業
01 Apr. 2021 - 31 Mar. 2024
姚 閔, 尾瀬 農之
日本学術振興会, 基盤研究(B), 北海道大学, 21H01754

Antibodies targeting dynamic epitopes generated by posttranslational protein glycosylation
Grants-in-Aid for Scientific Research
01 Apr. 2019 - 31 Mar. 2023
西村 紳一郎, 田中 良和, 比能 洋, 尾瀬 農之
本研究課題は、申請者らの独創的な「動的エピトープを標的とする新薬開発」という発想と、既にその基盤が完成しており効率的でシームレスな抗体開発までを可能とする「糖鎖工学プラットフォームを活用する戦略」によって創出されたED抗体（epitope-defined antibody）を用いて、主としてがん領域におけるアンメットメディカルニーズに応え得る新たながん治療薬の開発を目的としている。
この研究課題においては多くのがん細胞表面に高発現してがんの増殖や転移の促進などに深く関与することが広く知られるムチンMUC1の細胞外タンデムリピートドメインに生成する多様な動的エピトープ（dynamic epitope）を特異的な標的とするED抗体である抗MUC1モノクローナル抗体の構造改変と低分子化によって高機能化することで新たな創薬モダリティーを創出することを具体的な目標として進めている。
2021年度（３年目）はMUC1-STを動的エピトープとするSN131のFab-MUC1-ST共結晶の構造解析に成功した。また、物理化学的・生化学的キャラクタリゼーションに加えて、すい臓がん細胞表面のMUC1との結合やSiglecなどとの相互作用などを多角的に解析した（論文投稿準備中）。SN131の改変型抗体であるSN132（動的エピトープとの結合領域を含むanti-parallel coiled-coil構造のFv-claspとリン脂質DPC膜結合性membrane-associated peptide領域を融合した分子量40K程度の全く新しいタイプの機能性抗体フラグメント）の作成法を確立した（特許出願準備中）。
また、SN131およびSN121のFv領域を、2種類のリンパ球を認識する抗体（OKT3およびL2K）のFv領域と融合させて新たに８種の二重特異性抗体の作成を進めており高活性を示す高性能な二重特異性抗体取得に向けて研究が進展している。
Japan Society for the Promotion of Science, Grant-in-Aid for Scientific Research (A), Hokkaido University, 19H00918

生体分子中の窒素の特性を，中性子回折と計算化学により解明する　　　　　　　　　　　　　　　
Grants-in-Aid for Scientific Research Fund for the Promotion of Joint International Research (Fostering Joint International Research (B))
07 Feb. 2019 - 31 Mar. 2022
尾瀬 農之, 齋藤 徹, 塚本 卓
核酸やアミノ酸等の窒素化合物を代謝する過程においては，アミンのプロトタイプであるアンモニアが発生する。その反応性のために様々な細胞毒性を引き起こすアンモニアは，代謝や輸送をおこなう上で体内で注意して取り扱われなければならない。血液中に誤って放出されたアンモニアは，脳血液関門を容易に通過し脳障害をも引き起こす。陸上動物はふんだんに水を使用できないため，爬虫類・鳥類では尿酸として，両生類・哺乳類では尿素としてアンモニアを排出する系を備えている。これら排出回路を含め，アンモニアを扱う酵素には特別な設計がなされているおり，これの破綻は病気につながる。例えば，カルバモイルリン酸合成酵素(CPS)，シチジン３リン酸合成酵素(CTS)，グルタミン依存型アミドトランスフェラーゼ(Gat)を始めとし，アンモニアを反応中間体として利用する酵素は，求核性の高いアミンであるアンモニアが外部（細胞質）に漏れないよう，活性部位間がトンネルで結ばれている。トンネル内でアンモニアがどのように輸送されているかは，非常に興味深いトピックである。疎水性トンネルではアンモニアのまま輸送されていると予測される。また，親水性トンネルではプロトン化されていると予想されるが，アミンの性質を使用し，プロトン化と脱プロトン化を繰り返しながら輸送されているかもしれない。いずれにしても，Ｘ線結晶解析では水分子とアンモニア分子の違いを見分けることができないため，実験科学はデッドエンドになっていた。本提案では中性子線回折の利用方法をさらに進歩させ，窒素原子の持つ特徴を活かした構造生物学へと拡張する。アンモニアの分子内トンネルを使用した輸送機構は古細菌，真正細菌，真核生物に普遍的に見られる現象であるが，分子メカニズムの詳細は謎のままである。とりわけ，輸送に必要な水との協調作用を知る上では水分子と明確に区別する必要がある。
Japan Society for the Promotion of Science, Fund for the Promotion of Joint International Research (Fostering Joint International Research (B)), Hokkaido University, Principal investigator, Competitive research funding, 18KK0194

生体分子中におけるアミンの量子特性を解明する　　　　　　　　　　　　　　　
さきがけ
Oct. 2018 - Mar. 2022
尾瀬農之
JST, Principal investigator, Competitive research funding

必要時に可逆的立体構造形成する新規ペア型エーテル環化酵素の解析と再設計による応用　　　　　　　　　　　　　　　
Grants-in-Aid for Scientific Research Grant-in-Aid for Scientific Research on Innovative Areas (Research in a proposed research area)
01 Apr. 2019 - 31 Mar. 2021
尾瀬 農之
Japan Society for the Promotion of Science, Grant-in-Aid for Scientific Research on Innovative Areas (Research in a proposed research area), Hokkaido University, 19H04634

Elucidation of a tRNA conversion mechanism of transsulfursome by the structural analysis of quaternary complex
Grants-in-Aid for Scientific Research
01 Apr. 2017 - 31 Mar. 2020
Yao Min
To translate the protein according to the genetic code on the ribosome, the aminoacyl-tRNA (aa-tRNA) must be synthesized correctly. Generally, the aminoacyl-tRNA synthetase (aaRS) directly catalyzes this synthesis. However, in methanogenic archaea, a ternary complex called transsulfursome (SepRS, SepCys, SepCysS) synthesizes Cys-tRNA(Cys) in an indirect pathway.
In this study, we analyzed the relationship of structure and function of transsulfursome-tRNA complex by using X-ray crystallography, small-angle X-ray scattering, electron microscopic observation, and biochemical methods. Taken results together, we understood the dynamic molecular mechanism for the Cys-tRNA(Cys) synthesis of the transsulfursome.
Japan Society for the Promotion of Science, Grant-in-Aid for Scientific Research (B), Hokkaido University, 17H03637

JAK-STAT経路を不活化するため，ウイルスが採用する様々な戦略の分子機構解析　　　　　　　　　　　　　　　
Grants-in-Aid for Scientific Research Grant-in-Aid for Scientific Research (C)
01 Apr. 2017 - 31 Mar. 2020
尾瀬 農之
今回私達は, ITC測定を用いてMeV-VがSTAT1よりSTAT2に対して特異性が高いことを明らかにした。そこで，STAT2を主な標的としてType I IFNシグナル経路を阻害していると考えた。STAT2は, Type I IFNシグナル経路において, リン酸化, ヘテロ複合体化, 核移行, 転写活性化という過程を経る。この過程の中でMeV-VがSTAT2に干渉してシグナル伝達を阻害することを考えると, メカニズムとして リン酸化阻害， ヘテロ複合体化阻害，核移行阻害，転写活性化阻害が考えられた。
リン酸化阻害について, STAT2のリン酸化前後でMeV-Vの結合親和性が変化するか検証することを考え, 大腸菌TKB1でSTAT2を調製することを試みた。しかし, STAT1と同じ精製法でSTAT2を精製することはできなかった。また, リン酸化させたSTAT2は非常に凝集しやすい性質であった。バッファーや精製条件の検討により, リン酸化STAT2の調製を試みたいと考えている。したがってMeV-VのSTAT2に対する結合がリン酸化阻害に関わっているか不明であるが, Type I IFN経路阻害メカニズムとして現在多くの研究者に受け入れられているのは, MeV-VによるSTAT1やSTAT2のリン酸化阻害である（Chambers R. et al., 2009; Audsley M. D. et al., 2013）。私達が組換え調製したpY-STAT1を使ったITCによる相互作用解析では, MeV-Vとの相互作用が観測できなかった。これは, 非リン酸化STAT1の時に露出していた相互作用面がリン酸化に伴うコンホメーション変化によって隠れてしまったものと考えられ, MeV-Vはリン酸化を受ける前のSTAT1に対して特異的に結合する。
Japan Society for the Promotion of Science, Grant-in-Aid for Scientific Research (C), Hokkaido University, 17K07296

JAK-STAT経路を不活化するため，ウイルスが採用する様々な戦略の分子機構解析　　　　　　　　　　　　　　　
Apr. 2017 - Mar. 2020
尾瀬農之
科学研究費補助金（基盤C）, Principal investigator, Competitive research funding

４者複合体の構造解析によるtranssulfursomeのｔＲＮＡ変換機構の解明　　　　　　　　　　　　　　　
Apr. 2017 - Mar. 2020
姚閔
科学研究費補助金（基盤B）, Competitive research funding

Molecular mechanism analysis of STAT activation in breast tumor cells regulated by adaptor protein
Grants-in-Aid for Scientific Research
01 Apr. 2014 - 31 Mar. 2017
OSE Toyoyuki
The non-receptor tyrosine kinase breast tumor kinase (Brk), has been identified as a highly expressed Protein-tyrosine kinases in human melanocyte. Brk was reported to play key roles for cancer progression via overactivation of Signal Transducers and Activator of Transcription (STAT), especially STAT3 and STAT5. This activation is mediated by an adaptor protein STAP-2, which expresses ubiquitously and harobors many important functions for cell signal regulation. We successfully expressed recombinant Brk and STAP-2 including their mutants, checked kinase activity, monitored interactions, and structurally analyzed using small angle x-ray scattering (SAXS). The C-terminal phosphorylation site of Brk is, on the contrary to our expectation, didn’t produce open/close conformational change.
Japan Society for the Promotion of Science, Grant-in-Aid for Scientific Research (C), Hokkaido University, 26440016

アダプタータンパク質が担う，乳がん細胞における転写因子STATの活性化基盤の解明　　　　　　　　　　　　　　　
Apr. 2014 - Mar. 2017
尾瀬農之
科学研究費補助金（基盤C）, Principal investigator, Competitive research funding

Molecular basis regulating multiple signals by an adaptor protein
Grants-in-Aid for Scientific Research
01 Apr. 2012 - 31 Mar. 2014
OSE TOYOYUKI
The non-receptor tyrosine kinase breast tumor kinase (Brk), also known as PTK6, has been identified as a highly expressed Protein-tyrosine kinases in human melanocyte.
STAP-2 is also known as breast tumor kinase (Brk) substrate (BKS). it is important to better understand the contribution of Brk kinase activity and protein interactions to the STAT3-mediated signal transduction pathways in breast cancer. Brk phosphorylates STAP-2 and phosphorylated STAP-2 is believed to play an important role in Brk-mediated STAT3 activation. We purified Brk, STAP-2, and STAT3 and checked the kinase activity in vitro. Using purified proteins, we also checked the binding affinity between Brk and STAP-2. The information obtained by small angle xray scattering (SAXS) experiments was also useful to judge whether the conformational change of Brk is included (open/closed).
Japan Society for the Promotion of Science, Grant-in-Aid for Young Scientists (B), Hokkaido University, Principal investigator, Competitive research funding, 24770087

Elucidation of molecular mechanism of rice blast resistance
Grants-in-Aid for Scientific Research
01 Apr. 2011 - 31 Mar. 2014
SONE Teruo, TERAUCHI Ryohei, OSE Toyoyuki
Rice blast is the most important disease or rice. Blast is usually controlled by agrochemicals and resistant cultivars, but knowledge about molecular mechanisms of rice resistance toward blast is limited. This study aimed to elucidate the molecular mechanism of rice blast resistance by analyzing interaction of rice blast resistance gene Pia and blast virulence gene AVR-Pia. Results of this study indicated that AVR-Pia gene is started to be expressed before the penetration into rice cells, and after the secretion, AVR-Pia protein forms homodimer, and detected by rice Pia gene product. This study will contribute to further understanding of rice-rice blast interaction, which is the first step of resistance induction.
Japan Society for the Promotion of Science, Grant-in-Aid for Scientific Research (B), Hokkaido University, 23380024

Analysis of translation mechanism for selenoprotein synthesis
Grants-in-Aid for Scientific Research
2010 - 2011
OSE Toyoyuki
We successfully prepared protein and RNA molecules which are required for selecocystein incorporation on ribosome. These molecules are SBP2, L30, EFSec, SEICIS-RNA, tRNASec and subjected to biochemical and biophysical experiments. Crystal structure of L30 could be analyzed and published. Since we originally succeeded generating functional tRNASec, binding assay among tRNASec, EFSec, and SBP2 in the presence of specific amino acid are now in progress. We observed the significant binding affinity(K_d=229 nM) between the wild type and GDP using ITC, the way to prepare the molecule was thus confirmed. Furthermore, mutants generated for the purpose of crystallization were proved that they have proper secondary structures
Japan Society for the Promotion of Science, Grant-in-Aid for Young Scientists (B), Hokkaido University, Principal investigator, Competitive research funding, 22770093

第3の可変的リンパ球レセプターの構造と機能:リンパ球サブセットの起源
Grants-in-Aid for Scientific Research Grant-in-Aid for Scientific Research on Priority Areas
2010 - 2011
笠原 正典, 尾瀬 農之
獲得免疫系と、それを支えるリンパ球は長い間、有顎類に固有であると考えられてきた。ところが、leucine-rich repeatモジュールを組み換えることにより10^<10>を超える多様性を生み出す抗原レセプターである可変性リンパ球レセプター(variable lymphocyte receptor、以下VLR)が無顎類ヤツメウナギで発見されるに及んで、この考えは根底から覆されるに至った。先に、われわれは、VLRにAとBが存在することを示したが、最近、VLRAとBはそれぞれT、B様のリンパ球に発現され、Aは膜結合型レセプターとして、Bは抗体として機能することが示された。以上から、無顎類の段階で、細胞性免疫、液性免疫を担うリンパ球のサブセットがすでに存在することが明らかになった。今回、我々は第3のVLRを発見し、それをVLRCと命名した。本年度の研究により、1)VLRCはVLA+細胞、VLRB+細胞とは別のリンパ球サブセットに発現されていること、2)VLRCは分泌されないこと、3)VLRCは配列的には、VLRBに対してよりVLRAに類似性が高いこと、4)まれに単一リンパ球においてVLRAとVLRCの両者の組み換えが起きるが、一方のみが機能的なVLRタンパク質をコードできることなどが明らかになった。おそらく、VLRC+細胞はVLRA+細胞と同様、T細胞系の細胞と想定される。したがって、有顎類に3種のリンパ球サブセット(B、αβT、γδT細胞)が存在するように、無顎類にも3種のリンパ球サブセットが存在し、そのうちの二つはT細胞様で、一つはB細胞様であることが示唆された。研究分担者はVLRCの結晶を得て、その構造解析に成功した。
Japan Society for the Promotion of Science, Grant-in-Aid for Scientific Research on Priority Areas, Hokkaido University, Coinvestigator not use grants, Competitive research funding, 22021001

発熱現象を担う哺乳類の脱共役蛋白質の機能・構造解析
Grants-in-Aid for Scientific Research Grant-in-Aid for JSPS Fellows
2005 - 2005
尾瀬 農之
Japan Society for the Promotion of Science, Grant-in-Aid for JSPS Fellows, Kyushu University, 05J05880

エチレン前駆体ACCを分解する酵素反応の解析〜立体構造からのアプローチ〜
Grants-in-Aid for Scientific Research Grant-in-Aid for JSPS Fellows
2000 - 2002
尾瀬 農之
炭素-炭素結合を協奏的に形成する反応を触媒する酵素(macrophomate synthase)の立体構造解析に成功した。得られた立体構造から、このステップはDiels-Alder反応の可能性が極めて高くなった。Diels-Alder反応は、シクロヘキセンを合成する有機化学上極めて重要な反応である。この反応を触媒する酵素の立体構造は世界初のものであり、Nature誌で発表した。活性部位は巧妙に設計されており、より効率的にDiels-Alder反応を進行させるための戦略を見ることができる。基質を導入した活性部位の構造及び、変異実験や反応中間体の絶対配置に関する情報を組み合わせ、1)より活性化した基質を用いること、2)水素結合によって反応性を大きく上昇させること、3)生成物阻害を回避する構造変化を伴うこと、4)柔軟性に富んだループ領域をもっていることなどを具体的に明らかにした。この活性部位は、新たな化合物を作るための、抗体触媒を設計する上で非常に重要な情報となるものである。
三員環アミノ酸ACC(1-aminocyclopropane-1-carboxylate)を分解できる特殊な酵素ACC deaminase(yACCD;酵母Hansenula saturunus由来)の結晶構造を、分解能2.0Åで精密化した。立体構造および構造情報から予測した反応機構を、J.Biol.Chem.誌上に発表した。この反応機構を検証するために変異体を9種類作製した。活性消失が確認された変異体のうち、K51T, Y295FとACCを結合させた複合体の結晶化に成功し、反応中間体構造を得た。反応中でビタミンB6は基質と結合し、回転することで反応に最適な立体環境を提供する。活性残基Lys51はACCのメチレン側鎖からプロトンを引き抜くモデルを提唱することができた。これはビタミンB6酵素では初のものであり、進化の多様性を表すモデルと言える。論文投稿中である。また、超高度好熱古細菌P.horikoshiiにおいてACCDとアサインされた遺伝子(PH0054)がコードする蛋白質の、ACC存在下での構造解析にも成功し、大きなドメイン変化が観察できた。ACCD活性の発現は当初の予想以上に厳密な環境制御を要している。論文投稿準備中である。
Japan Society for the Promotion of Science, Grant-in-Aid for JSPS Fellows, Hokkaido University, 00J07161