分子シャペロンマシーナリーのX線結晶構造解析

• Project/Area Number: 11169213
• Research Category: Grant-in-Aid for Scientific Research on Priority Areas (A)
• Allocation Type: Single-year Grants
• Review Section: Biological Sciences
• Research Institution: Tokyo Institute of Technology
• Principal Investigator: 田口 英樹 東京工業大学, 資源化学研究所, 助手 (40272710)
• Project Period (FY): 1999 – 2000
• Project Status: Completed (Fiscal Year 2000)
• Budget Amount: ¥3,400,000 (Direct Cost: ¥3,400,000); Fiscal Year 2000: ¥1,700,000 (Direct Cost: ¥1,700,000); Fiscal Year 1999: ¥1,700,000 (Direct Cost: ¥1,700,000)
• Keywords: 分子シャペロン / GroEL / 熱ショックタンパク質 / フォールディング

Research Abstract

細胞内でのタンパク質の一生のケアを行う分子シャペロンは、そのほとんどが複雑なオリゴマー構造を取ることで機能する。立体構造未知の分子シャペロンの構造解析に向けて、本年度は、結晶解析に適している好熱菌の分子シャペロンの大量発現系の構築および機能解析を中心に研究を進めた。
扱ったシャペロンは以下の通りである。
1)好熱菌ClpXPプロテアーゼ複合体(プロテアソーム・ホモログ)
2)好熱菌ClpB(Hsp104ホモログ)
3)好熱菌FtsHプロテアーゼ
4)好熱菌DnaK/DnaJ/DafA複合体

Research Products

• [Publications] Shiseki,K.: "Synchronized Domain Opening Motion of GroEL is Essential for Communication between the Two Rings."J.Biol.Chem.. 276(In press). (2001)
• [Publications] Yokoyama,K.: "V-Type H+-ATP ase/synthase from a Thermophilic Eubacterium, Thermus Thermophilus, Subunit Structure And Operon"J.Biol.Chem.. 275. 13955-13961 (2000)
• [Publications] Watanabe,Y.: "Heat-inactivated proteins managed by DnaKJ-GrpE-GlpB chaperones are released as a chaperonin-recognizable nonnative form."J.Biol.Chem.. 275. 12388-12392 (2000)
• [Publications] Asahara,Y.: "FtsH recognizes unfolded proteins and hydrolyzes carboxyl side of hydrophobic residues."J.Biochem.. 127. 931-937 (2000)
• [Publications] Pack,C-G.: "Effect of electrostatic interactions on the binding of charged substrate to GroEL studied by highly sensitive fluorescence correlation spectroscopy"Biochem.Biophys.Res.Commun.. 267. 300-304 (2000)
• [Publications] Aoki,K.: "GroEL binds artificial proteins with random sequences."J.Biol.Chem.. 275. 13755-13758 (2000)
• [Publications] 田口英樹(分担): "シャペロニンとGFPフォールディング"生物工学会誌. 3 (2000)
• [Publications] 田口英樹(分担): "タンパク質のケアテイカー、分子シャペロン"共立出版. 19 (2000)
• [Publications] Aoki, K.: "GroEL binds artificial proteins with random sequences"J. Biol. Chem.. (in press). (2000)
• [Publications] Pack, C-G.: "Effect of electrostatic interactions on the binding of charged substrate to GroEl studied by highly sensitive fluorescence correlation spectroscopy"Biochem. Biophys. Res. Commum.. 267. 300-304 (2000)
• [Publications] Sakikawa, C: "On the maximum size of proteins to stay and fold in the cavity of GroEL underneath GroES"J. Biol. Chem.. 274. 21251-21256 (1999)
• [Publications] Pack, C-G.: "Analysis of interaction between chaperonin GroEL and its substrate using fluorescence correlation spectroscopy"Cytometry. 36. 247-253 (1999)