MISC

基本情報

氏名 古賀 雄一
氏名(カナ) コガ ユウイチ
氏名(英語) Koga Yuuichi
所属 工学部 応用化学科
職名 教授
researchmap研究者コード 5000076449
researchmap機関 岡山理科大学

題名

Four new crystal structures of Tk-subtilisin in unautoprocessed, autoprocessed and mature forms: Insight into structural changes during maturation

単著・共著の別

 

著者

Shun-ichi Tanaka
Hiroyoshi Matsumura
Yuichi Koga
Kazufumi Takano
Shigenori Kanaya

概要

Subtilisin from the hyperthermophilic archaeon Thermococcus kodakaraensis (Tk-subtilisin) is matured from Pro-Tk-subtilisin upon autoprocessing and degradation of the propeptide. The crystal structures of the autoprocessed and mature forms of Tk-subtilisin were determined at 1.89 angstrom and 1.70 angstrom resolution, respectively. Comparison of these structures with that of unautoprocessed Pro-Tk-subtilisin indicates that the structure of Tk-subtilisin is not seriously changed during, maturation. However, one unique Ca2+-binding site (Ca-7) is identified in these structures. In addition, the N-terminal region of the mature domain (Gly70-Pro82), which binds tightly to the main body in the unautoprocessed form, is disordered and mostly truncated in the autoprocessed and mature forms, respectively. Interestingly, this site is formed also in the unautoprocessed form when its crystals are soaked with 10 mM CaCl2, as revealed by the 1.87 angstrom structure. Along with the formation of this site, the N-terminal region (Leu75-Thr80) is disordered, with the scissile peptide bond contacting with the active site. These results indicate that the calcium ion binds weakly to the Ca-7 site in the unautoprocessed form, but is trapped upon autoprocessing. We propose that the Ca-7 site is required to promote the autoprocessing reaction by stabilizing the autoprocessed form, in which the new N terminus of the mature domain is structurally disordered. Furthermore, the crystal structure of the Tk-propeptide:S324A-subtilisin complex, which was formed by the addition of separately expressed proteins, was determined at 1.65 angstrom resolution. This structure is virtually identical with that of the autoprocessed form, indicating that the interaction between the two domains is highly intensive and specific. (c) 2007 Elsevier Ltd. All rights reserved.

発表雑誌等の名称

JOURNAL OF MOLECULAR BIOLOGY

出版者

ACADEMIC PRESS LTD- ELSEVIER SCIENCE LTD

372

4

開始ページ

1055

終了ページ

1069

発行又は発表の年月

2007-09

査読の有無

無し

依頼の有無

無し

記述言語

英語

掲載種別

 

ISSN

 

ID:DOI

10.1016/j.jmb.2007.07.027

ID:NAID(CiNiiのID)

 

ID:PMID

 

JGlobalID

 

arXiv ID

 

ORCIDのPut Code

 

DBLP ID