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

大腸菌蛋白質分泌の分子機構

Research Project

Project/Area Number 03264102
Research Category

Grant-in-Aid for Scientific Research on Priority Areas

Allocation TypeSingle-year Grants
Research InstitutionThe University of Tokyo

Principal Investigator

水島 昭二  東京大学, 応用微生物研究所, 教授 (50013313)

Co-Investigator(Kenkyū-buntansha) 伊藤 維昭  京都大学, ウイルス研究所, 教授 (90027334)
Project Period (FY) 1991
Project Status Completed (Fiscal Year 1991)
Budget Amount *help
¥28,800,000 (Direct Cost: ¥28,800,000)
Fiscal Year 1991: ¥28,800,000 (Direct Cost: ¥28,800,000)
Keywords蛋白質の分泌 / 蛋白質の膜透過 / Sec蛋白質 / SecY / SecE / シグナルペプチド / 変異解析 / SecA
Research Abstract

1.生化学を中心とした研究(水島グル-プで行われた研究)
(1)精製したSecA,SecE,SecYより膜透過活性を再構成することに成功した。SecD,SecFも精製したが、これらは再構成膜透過活性を促進しなかった。
(2)SecEのC末端領域がSecYとの相互作用に関与していること、C末端領域のみでSecE活性を示すことを見い出した。
(3)再構成された活性はATP依存性であった。しかし、プロトン駆動力依存性を見ることはできなかった。
(4)シグナルペプチドのN末端正荷電と中央疎水領域が機能的に関連しあっていることを見い出した。
(5)タンパク質膜透過装置は直鎖ポリペプチド鎖以外の残基をも許容しうることを見い出した。
2.遺伝学を中心とした研究(伊藤グル-プで行われた研究)
(1)優性欠損SecY変異(SecY^<ーd>1:細胞質領域5の3アミノ酸基欠失)および多数のCs分泌欠損secY変異を分離し、塩基配列変化を決定した。
(2)SecY^<ーd>1の分泌阻害を回復させるsecY内部のリンカ-挿入変異の分離マルチコピ-状態で回復させるsecY以外の遺伝子領域ydrの同定を行った。
(3)アルカリ性ホスファタ-ゼのSS結合の形成・foldingに必要な新たな遺伝子ppfAを同定した。
(4)膜タンパク質の「透過停止配列」のC末端側に融合したPhoAを膜透過させてしまう変異stdを分離し、それが遺伝子ftsHの変異である事を示した。

Report

(1 results)
  • 1991 Annual Research Report
  • Research Products

    (14 results)

All Other

All Publications (14 results)

  • [Publications] Ken-ichi Nishiyama: "SecY is an indispensable component of the protein secretory machinery of Escherichia coli." Biochim.Biophys.Acta. 1065. 89-97 (1991)

    • Related Report
      1991 Annual Research Report
  • [Publications] Jiro Akimaru: "Reconstitution of a protein secretory machinery from purified SecY,SecE,and SecA of Escherichia coli." Proc.Natl.Acad.Sci.USA. 88. 6545-6549 (1991)

    • Related Report
      1991 Annual Research Report
  • [Publications] Katsuko Tani: "A chemically cross-linked nonlinear proOmoA molecule can be translocated into everted membrane vesicles of Escherichia coli in the presence of the proton motive force." FEBS Letters. 285. 127-131 (1991)

    • Related Report
      1991 Annual Research Report
  • [Publications] Shoji Mizushima: "In vitro biochemical studies on translocation of presecretory proteins across the cytoplasmic membrane of Escherichia coli." Methods in Cell Biol.34. 107-146 (1991)

    • Related Report
      1991 Annual Research Report
  • [Publications] Masashi Kato: "In Vitro translocation of secretory proteins possessing no charges at the mature domain takes place efficiently in a protonmotive" J.Biol.Chem.267. 413-418 (1992)

    • Related Report
      1991 Annual Research Report
  • [Publications] Chinami Hikita: "Effects of total-hydrophobicity and length of the hydrophobic domain of a signal peptide on in vitro translocation efficiency." J.Biol.Chem.(1992)

    • Related Report
      1991 Annual Research Report
  • [Publications] Ken-ichi Nishiyama: "The C-terminal region of SecE interacts with SecY and is functional in the reconstitution of protein translocation activity in Escherichia coli." J.Biol.Chem.(1992)

    • Related Report
      1991 Annual Research Report
  • [Publications] Chinami Hikita: "The requirement of a positive charge at the amino-terminus can be compensated for by a longer central hydrophobic stretch in the functioning of signal peptides." J.Biol.Chem.(1992)

    • Related Report
      1991 Annual Research Report
  • [Publications] Sakaguchi,M.,Ueguchi,C.,Ito,K.,and Omura,T.: "Yeast gene which suppresses the defect in protein export of a secY mutant of E.coli." J.Biochem.109. 799-802 (1991)

    • Related Report
      1991 Annual Research Report
  • [Publications] Taura,T.,and Ito,K.: "Does protein secretion activity vary during the cell cycle of Escherichia coli?" J.Biochem.109. 811-815 (1991)

    • Related Report
      1991 Annual Research Report
  • [Publications] Ito,K.and Akiyama,Y.: "In vivo analysis of integration of membrane proteins in Escherichia coli." Mol.Microbiol.5. 2243-2253 (1991)

    • Related Report
      1991 Annual Research Report
  • [Publications] Kamitani,S.,Akiyama,Y.,and Ito,K.: "Identification and characterization of an Escherichia coli gene required for the formation of correctly folded alkaline phosphatase,a periplasmic enzyme." EMBO J.11. 57-62 (1992)

    • Related Report
      1991 Annual Research Report
  • [Publications] Ueguchi,C.,and Ito,K.: "Multicopy suppression:an approach to understanding intracellular functioning of the protein export system." J.Bacteriol.174. (1992)

    • Related Report
      1991 Annual Research Report
  • [Publications] Shimoike,T.,Akiyama,Y.,Baba,T.,Taura,T.,and Ito,K.: "SecY variants that interfere with Escherichia coli protein export in the presence of normal secY." Mol.Microbiol.6. (1992)

    • Related Report
      1991 Annual Research Report

URL: 

Published: 1991-04-01   Modified: 2016-04-21  

Information User Guide FAQ News Terms of Use Attribution of KAKENHI

Powered by NII kakenhi