Structural Study on Mental Disease Gene Product Protein

• Project/Area Number: 15K06980
• Research Category: Grant-in-Aid for Scientific Research (C)
• Allocation Type: Multi-year Fund
• Section: 一般
• Research Field: Structural biochemistry
• Research Institution: Hosei University (2016-2017); Yokohama City University (2015)
• Principal Investigator: UNZAI Satoru 法政大学, 生命科学部, 准教授 (60336592)
• Co-Investigator(Renkei-kenkyūsha): PARK Sam-Yong 横浜市立大学, 生命医科学研究科, 教授 (20291932)
• Research Collaborator: KOMIYAMA Noboru The University of Edinburgh, Centre for Clinical Brain Sciences, Senior Lecturer
• Project Period (FY): 2015-04-01 – 2018-03-31
• Project Status: Completed (Fiscal Year 2017)
• Budget Amount: ¥5,200,000 (Direct Cost: ¥4,000,000、Indirect Cost: ¥1,200,000); Fiscal Year 2017: ¥1,560,000 (Direct Cost: ¥1,200,000、Indirect Cost: ¥360,000); Fiscal Year 2016: ¥1,820,000 (Direct Cost: ¥1,400,000、Indirect Cost: ¥420,000); Fiscal Year 2015: ¥1,820,000 (Direct Cost: ¥1,400,000、Indirect Cost: ¥420,000)
• Keywords: タンパク質 / 結晶構造解析 / X線結晶構造解析 / シグナル伝達 / Rap1b / SynGAP

Outline of Final Research Achievements

The brain-specific synaptic GTPase activating protein (SynGAP) is important in synaptic plasticity. Mutations in SYNGAP1 gene cause epileptic encephalopathy, intellectual disability, and autism via haploinsufficiency. Basic research is needed to better understand the molecular functions of the SynGAP protein, which will lead to targets for therapeutic intervention. Structural biology methods were applied for the SynGAP function research. Recombinant SynGAP and Rap1B (SynGAP can enhance Rap1B GTPase activity) were prepared using e-coli expression system. We succeeded in solving high resolution crystal structure of the Rap1B protein and understanding its molecular switch mechanism. We established small volume GTPase activity measurement system using malachite green. Rap1B GTPase activity enhancement by SynGAP can be detected by this system.

Research Products

• [Int'l Joint Research] University of Edinburgh(英国)
• [Int'l Joint Research] University of Edinburgh(英国)
• [Journal Article] Analytical ultracentrifugation in structural biology (2018)
  • Author(s): Unzai Satoru
  • Journal Title: Biophysical reviews Volume: 10 Issue: 2 Pages: 229-233
  • DOI: 10.1007/s12551-017-0340-0
  • Peer Reviewed / Open Access
• [Journal Article] Molecular Landscape of the Ribosome Pre-initiation Complex during mRNA Scanning: Structural Role for eIF3c and Its Control by eIF5 (2017)
  • Author(s): Obayashi E, Luna RE, Nagata T, Martin-Marcos P, Hiraishi H, Singh CR, Erzberger JP, Zhang F, Arthanari H, Morris J, Pellarin R, Moore C, Harmon I, Papadopoulos E, Yoshida H, Nasr ML, Unzai S, Thompson B, Aube E, Hustak S, Stengel F, Dagraca E, Ananbandam A, Gao P, Urano T, Hinnebusch AG, Wagner G, Asano K.
  • Journal Title: Cell Report Volume: 18 Issue: 11 Pages: 2651-2663
  • DOI: 10.1016/j.celrep.2017.02.052
  • NAID: 120006358788
  • Peer Reviewed / Open Access / Int'l Joint Research / Acknowledgement Compliant
• [Journal Article] Structural insights into a 20.8-kDa tegumental-allergen-like (TAL) protein from Clonorchis sinensis (2017)
  • Author(s): Jo Chang Hwa、Son Jonghyeon、Kim Sulhee、Oda Takashi、Kim Jaehoon、Lee Myoung-Ro、Sato Mamoru、Kim Hyun Tae、Unzai Satoru、Park Sam-Yong、Hwang Kwang Yeon
  • Journal Title: Scientific Reports Volume: 7 Issue: 1 Pages: 1764-1764
  • DOI: 10.1038/s41598-017-02044-0
  • Peer Reviewed / Open Access / Int'l Joint Research
• [Journal Article] Heat-induced native dimerization prevents amyloid formation by variable domain from immunoglobulin light-chain REI (2017)
  • Author(s): Nawata Marina、Tsutsumi Hirotaka、Kobayashi Yuta、Unzai Satoru、Mine Shouhei、Nakamura Tsutomu、Uegaki Koichi、Kamikubo Hironari、Kataoka Mikio、Hamada Daizo
  • Journal Title: The FEBS Journal Volume: 284 Issue: 18 Pages: 3114-3127
  • DOI: 10.1111/febs.14181
  • Peer Reviewed / Open Access
• [Journal Article] Oligomerization of Hmo1 mediated by box A is essential for DNA binding in vitro and in vivo. (2016)
  • Author(s): K. Kasahara, A. Higashino, S. Unzai, H. Yoshikawa, T. Kokubo
  • Journal Title: Genes Cells Volume: 21(12) Issue: 12 Pages: 1333-1352
  • DOI: 10.1111/gtc.12449
  • Peer Reviewed / Open Access
• [Journal Article] Crystal structure of MytiLec, a galactose-binding lectin from the mussel Mytilus galloprovincialis with cytotoxicity against certain cancer cell types. (2016)
  • Author(s): Terada D, Kawai F, Noguchi H, Unzai S, Hasan I, Fujii Y, Park SY, Ozeki Y, Tame JRH
  • Journal Title: Sci Rep Volume: 6 Issue: 1 Pages: 28344-28344
  • DOI: 10.1038/srep28344
  • Peer Reviewed / Open Access / Int'l Joint Research / Acknowledgement Compliant
• [Journal Article] (9)Structural insight into photoactivation of an adenylate cyclase from a photosynthetic cyanobacterium (2016)
  • Author(s): Ohki M, Sugiyama K, Kawai F, Tanaka H, Nihei Y, Unzai S, Takebe M, Matsunaga S, Adachi S, Shibayama N, Zhou Z, Koyama R, Ikegaya Y, Takahashi T, Tame J, Iseki M, Park S-Y.
  • Journal Title: Proc Natl Acad Sci USA Volume: 113 Issue: 24 Pages: 6689-6664
  • DOI: 10.1073/pnas.1517520113
  • Peer Reviewed / Open Access / Int'l Joint Research / Acknowledgement Compliant
• [Journal Article] The structure and conformational switching of Rap1B (2015)
  • Author(s): H. Noguchi, T. Ikegami, A. Nagadoi, Y. O. Kamatari, S. Y. Park, J. R. H. Tame, S. Unzai
  • Journal Title: Biochem Biophys Res Commun. Volume: 印刷中 Issue: 1 Pages: 46-51
  • DOI: 10.1016/j.bbrc.2015.04.103
  • Peer Reviewed / Open Access
• [Journal Article] Self-Assembling Nano-Architectures Created from a Protein Nano-Building Block Using an Intermolecularly Folded Dimeric de Novo Protein. (2015)
  • Author(s): Kobayashi N, Yanase K, Sato T, Unzai S, Hecht MH, Arai R.
  • Journal Title: J Am Chem Soc. Volume: 137 Issue: 35 Pages: 11285-11293
  • DOI: 10.1021/jacs.5b03593
  • Peer Reviewed / Int'l Joint Research
• [Journal Article] A novel 3' splice site recognition by the two zinc fingers in the U2AF small subunit. (2015)
  • Author(s): Yoshida H, Park S-Y, Oda T, Akiyoshi T, Sato M, Shirouzu M, Tsuda K, Kuwasako K, Unzai S, Muto Y, Urano T, Obayashi E.
  • Journal Title: Genes Dev. Volume: 29 Issue: 15 Pages: 1649-1660
  • DOI: 10.1101/gad.267104.115
  • Peer Reviewed / Open Access
• [Journal Article] Crystal structure of human importin-α1 (Rch1), revealing a potential autoinhibition mode involving homodimerization (2015)
  • Author(s): Miyatake H., Sanjoh A., Unzaic S., Matsuda G., Tatsumi Y., Miyamoto Y., Dohmae N. and Aida Y.
  • Journal Title: PLoS One Volume: 10(2) Issue: 2 Pages: 1-14
  • DOI: 10.1371/journal.pone.0115995
  • Peer Reviewed / Open Access
• [Journal Article] A multilaboratory comparison of calibration accuracy and the performance of external references in analytical ultracentrifugation. (2015)
  • Author(s): Zhao H, et al
  • Journal Title: PLoS ONE. Volume: 10 Issue: 5 Pages: e0126420-e0126420
  • DOI: 10.1371/journal.pone.0126420
  • Peer Reviewed / Open Access / Int'l Joint Research
• [Presentation] Disassembly and Impact on Function of a Multimeric Allosteric Protein by Kosmotropes. (2017)
  • Author(s): Antonio Tsuneshige, Satoru Unzai
  • Organizer: 日本生物物理学会 第55回年会
• [Presentation] Molecular mechanism of the triplex DNA-binding protein to recognize triplex DNA (2016)
  • Author(s): Kazuki Kikuchi, Kohta Sugiyama, Ryotaro Kishi, Satoru Unzai, Hidetaka Torigoe
  • Organizer: 第54回日本生物物理学会
  • Place of Presentation: つくば国際会議場（茨城県つくば市）
  • Year and Date: 2016-11-25
• [Presentation] The structrue and conformational switching of Rap1b (2015)
  • Author(s): Noguchi, H., Park, S.-Y. Tame, J.R.H., and Unzai, S.
  • Organizer: 第53回生物物理学会年会
  • Place of Presentation: 金沢大学
  • Year and Date: 2015-09-15