Structural basis for the post translational membrane insertion by GET complex

• Project/Area Number: 24570126
• Research Category: Grant-in-Aid for Scientific Research (C)
• Allocation Type: Multi-year Fund
• Section: 一般
• Research Field: Structural biochemistry
• Research Institution: The University of Tokyo
• Principal Investigator: YAMAGATA ATSUSHI 東京大学, 放射光連携研究機構, 助教 (20463903)
• Project Period (FY): 2012-04-01 – 2015-03-31
• Project Status: Completed (Fiscal Year 2014)
• Budget Amount: ¥5,460,000 (Direct Cost: ¥4,200,000、Indirect Cost: ¥1,260,000); Fiscal Year 2014: ¥1,820,000 (Direct Cost: ¥1,400,000、Indirect Cost: ¥420,000); Fiscal Year 2013: ¥1,820,000 (Direct Cost: ¥1,400,000、Indirect Cost: ¥420,000); Fiscal Year 2012: ¥1,820,000 (Direct Cost: ¥1,400,000、Indirect Cost: ¥420,000)
• Keywords: 結晶構造解析 / テイルアンカー型タンパク質 / 膜タンパク質 / 構造生物学 / X線結晶解析 / TA タンパク質 / X線結晶構造解析

Outline of Final Research Achievements

Get3 is the specific targeting factor for the tail anchored proteins, which is defined by the C-terminal single transmembrane domain. After capturing the TA proteins, Get3 is further localized on the ER membrane through the interaction with Get1/Get2 membrane proteins. In this study, we succeeded to solve the crystal structure of the complex between Get3 and Get1 cytosolic domain. In addition, we succeeded to purify the Get3-TA complex coexpressed in E. coli. By using the mutant Get3, we succeeded to obtain the Get3-TA complex at a molar ratio of 2:1 (Get3 dimer bound to single TA). Further, we succeeded to purify Get1, Get2 using insect cell expression system.

Research Products

• [Journal Article] Structures of CYLD USP with Met1- or Lys63-linked diubiquitin reveal mechanisms for dual specificity (2015)
  • Author(s): Sato, Y., Goto, E., Shibata, Y., Kubota, Y., Yamagata, A., Goto-Ito, S., Kubota, K., Inoue, J., Takekawa, M., Tokunaga, F. and Fukai, S.
  • Journal Title: Nat. Struct. Mol. Biol. Volume: 22 Issue: 3 Pages: 222-229
  • DOI: 10.1038/nsmb.2970
  • Peer Reviewed / Open Access / Acknowledgement Compliant
• [Journal Article] Switching and emergence of CTL epitopes in HIV-1 infection (2014)
  • Author(s): Han, C., Kawana-Tachikawa, A., Shimizu, A., Zhu D., Nakamura, H., Adachi, E., Kikuchi, T., Koga, M., Koibuchi, T., Gao, G.F., Sato, Y., Yamagata, A., Martin, E., Fukai, S., Brumme, Z.L.,Iwamoto, A.
  • Journal Title: Retrovirolgy Volume: 11 Issue: 1 Pages: 38-38
  • DOI: 10.1186/1742-4690-11-38
  • Peer Reviewed / Open Access
• [Journal Article] Switching and emergence of CTL epitopes in HIV-1 infection (2014)
  • Author(s): Han C, Kawana-Tachikawa A, Shimizu A, Zhu D, Nakamura H, Adachi E, Kikuchi T, Koga M, Koibuchi T, Gao GF, Sato Y, Yamagata A, Martin E, Fukai S, Brumme ZL, Iwamoto A
  • Journal Title: Retrovirology Volume: ?
  • Peer Reviewed
• [Journal Article] Structure of TCR and antigen complexes at an immunodominant CTL epitope in HIV-1 infection (2013)
  • Author(s): Shimizu, A., Kawana-Tachikawa. A., Yamagata, A., Han, C., Zhu, D., Sato, Y., Nakamura, H., Koibuchi, T., Carlson, J., Martin, E., Brumme, C.J., Shi, Y., Gao, G.F., Brumme, Z.L., Fukai, S., Iwamoto, A.
  • Journal Title: Scientific Reports Volume: 3 Issue: 1 Pages: 3097-3097
  • DOI: 10.1038/srep03097
  • Peer Reviewed
• [Journal Article] Get1 stabilizes an open dimer conformation of get3 ATPase by binding two distinct interfaces. (2012)
  • Author(s): Kubota K, Yamagata A, Sato Y, Goto-Ito S, Fukai S.
  • Journal Title: J Mol Biol. Volume: 422(3) Issue: 3 Pages: 366-75
  • DOI: 10.1016/j.jmb.2012.05.045
  • Peer Reviewed
• [Journal Article] Molecular basis of Lys-63-linked polyubiquitination inhibition by the interaction between human deubiquitinating enzyme OTUB1 and ubiquitin-conjugating enzyme UBC13. (2012)
  • Author(s): Sato Y, Yamagata A, Goto-Ito S, Kubota K, Miyamoto R, Nakada S, Fukai S.
  • Journal Title: J Biol Chem. Volume: 287(31) Issue: 31 Pages: 25860-8
  • DOI: 10.1074/jbc.m112.364752
  • Peer Reviewed
• [Presentation] Bat3 の新規ドメインの構造解析 (2014)
  • Author(s): 山形敦史、佐藤裕介、伊藤（後藤）桜子、深井周也
  • Organizer: 日本結晶学会年会
  • Place of Presentation: 東京大学弥生講堂（東京都・文京区）
  • Year and Date: 2014-11-01 – 2014-11-03
• [Presentation] Structural basis for the post translational membrane insertion by Get3 (2013)
  • Author(s): 山形 敦史
  • Organizer: International conference on structural genomics
  • Place of Presentation: 札幌
• [Presentation] Structural Basis for the posttranslational membrane insertion by Get3 (2012)
  • Author(s): 山形 敦史
  • Organizer: 日本分子生物学会年会
  • Place of Presentation: 福岡国際会議場
• [Remarks] 東京大学放射光連携研究機構生命科学部門
  • URL: http://www.iam.u-tokyo.ac.jp/srro/SRROLifeSciDivJp2/Welcome.html
• [Remarks] 東京大学放射光連携研究機構生命科学部門
  • URL: http://www.iam.u-tokyo.ac.jp/srro/SRROLifeSciDivJp2/Welcome.html
• [Remarks] 東京大学放射光連携研究機構生命科学部門
  • URL: http://www.iam.u-tokyo.ac.jp/srro/SRROLifeSciDivJp2/Welcome.html