• Search Research Projects
  • Search Researchers
  • How to Use
  1. Back to previous page

Structure analysis of SecA-SecYE protein translocation machinery

Research Project

Project/Area Number 16570094
Research Category

Grant-in-Aid for Scientific Research (C)

Allocation TypeSingle-year Grants
Section一般
Research Field Structural biochemistry
Research InstitutionKyoto University

Principal Investigator

MORI Hiroyuki  Kyoto Univ., Institute for Virus Res., Assistant Professor, ウイルス研究所, 助手 (10243271)

Project Period (FY) 2004 – 2005
Project Status Completed (Fiscal Year 2005)
Budget Amount *help
¥3,800,000 (Direct Cost: ¥3,800,000)
Fiscal Year 2005: ¥1,900,000 (Direct Cost: ¥1,900,000)
Fiscal Year 2004: ¥1,900,000 (Direct Cost: ¥1,900,000)
KeywordsMembrane protein / Protein translocation / SecA / SecYE / Thermus thermophilus / crystal structure / X-ray / X線結晶構造解析 / SecY / SecE / ATPase / チャネル
Research Abstract

In bacteria, one third of newly synthesized proteins must be translocatedacrossthecytoplasmicmembranetolocalize their final destinations. In this reaction, SecA and SecYE complex have central roles. The former is an essential ATPase to drive the reaction and the latter is an evolutionally conserved membrane protein complex, which is thought to form a specific channel for a translocatingpolypeptide. However, molecular mechanisms of the reaction still remain poorly understood. In addition, bacteria have a multi-path membrane protein, SecDF, which form a translocon-associated complex required for efficient preprotein translocation and membrane protein integration.
In this study, we established over-expression in E.coli and purification systems of these Sec factors from Thermusthemophilus HB8, and crystallized their proteins. We succeeded in determination of 3 D structure of TSecA at 2.8A resolution. Contrary to previous reports, the structure revealed occurrence of a parallel dimer. Biochemical studies also confirmed this conclusion and supported that this parallel dimer was dominant in soution. In the case of TSecDF, we obtained complete data sets at 3.7 A resolution and started to build a molecular model of TSecDF. Although we obtained good-shape TSecYE crystals, they only diffracted X-rays to 6 A. To improve the diffraction quality of TSecYE crystals, we prepared monoclonal antibodies against TSecYE and selected one that stably bound to the TSecYE complex. Fab fragments prepared from the monoclonal antibody seem to be a good tool to stabilize the TSecYE complex in crystal lattice.

Report

(3 results)
  • 2005 Annual Research Report   Final Research Report Summary
  • 2004 Annual Research Report
  • Research Products

    (9 results)

All 2006 2005 2004

All Journal Article (9 results)

  • [Journal Article] Purification, crystallization and preliminary X-ray diffraction of SecDF, a translocon-associated membrane protein, from Thermus thermophilus2006

    • Author(s)
      Tsukazaki T, Mori H et al.(15名)
    • Journal Title

      Acta Crystallographica Section F. 62

      Pages: 376-380

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2005 Final Research Report Summary
  • [Journal Article] Purification, crystallization and preliminary X-ray diffraction of SecDF, a translocon-associated membrane protein, from Therm us thermophilus2006

    • Author(s)
      Tsukazaki, T., Mori, H., Fukai, S., Numata, T., Perederina, A., Adachi.H., Matsumura, H., Takano, K., Murakami, S., Inoue, T., Mori, Y., Sasaki, T., Vassylyev, D.G., Nureki, O., Ito, K.
    • Journal Title

      Acta Ciystallographica Section F 62

      Pages: 376-380

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      2005 Final Research Report Summary
  • [Journal Article] Purifiction, crystallization and preliminary X-ray diffraction of ScDF a translocon associated membrane protein, from Thermurs thermophilus2006

    • Author(s)
      Tsukazaki T, Mori H et al. (15名)
    • Journal Title

      Acta Crystallographica Section F 62

      Pages: 376-380

    • Related Report
      2005 Annual Research Report
  • [Journal Article] SecM facilitates translocase function of SecA by localizin its biosynthesis2005

    • Author(s)
      Nakatogawa H, Murakami A, Mori H, Ito K
    • Journal Title

      Genes & Development 19

      Pages: 436-444

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2005 Final Research Report Summary
  • [Journal Article] SecM facilitates translocase function of SecA by localizing its biosynthesis.2005

    • Author(s)
      Nakatogawa, H., Murakami, A., Mori, H., Ito, K.
    • Journal Title

      Genes & Development 19

      Pages: 436-444

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      2005 Final Research Report Summary
  • [Journal Article] SecM facilitates translocase function of SecA by localizing its biosynthesis2005

    • Author(s)
      Nakatogawa, H., Murakami, A., Hori, H., Ito, K.
    • Journal Title

      Genes and Development 19

      Pages: 436-444

    • Related Report
      2004 Annual Research Report
  • [Journal Article] Mutational analysis of transmembrane regions 3 and 4 of SecY a central component of protein translocase2004

    • Author(s)
      Mori H, Shimokawa N, Satoh Y, Ito K
    • Journal Title

      J. Bacteriol. 186

      Pages: 3960-3969

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2005 Final Research Report Summary
  • [Journal Article] Mutational analysis of transmembrane regions 3 and 4 of SecY, a central component of protein translocase2004

    • Author(s)
      Mori, H., Shimokawa, N., Satoh, Y., Ito, K.
    • Journal Title

      J.Bacteriol. 186

      Pages: 3960-3969

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      2005 Final Research Report Summary
  • [Journal Article] Mutational Analysis of Transmembrane Regions 3 and 4 of SecY, a Central Component of Protein Translocase.2004

    • Author(s)
      Mori, H., Shimokawa, N., Satoh, Y., Ito, K.
    • Journal Title

      Journal of Bacteriology 186

      Pages: 3960-3969

    • Related Report
      2004 Annual Research Report

URL: 

Published: 2004-04-01   Modified: 2016-04-21  

Information User Guide FAQ News Terms of Use Attribution of KAKENHI

Powered by NII kakenhi