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

Crystal structure of highly thermostable glycerol kinase from a hyperthermophilic archaeon in a dimeric form.

単著・共著の別

 

著者

Yuichi Koga
Ryota Katsumi
Dong-Ju You
Hiroyoshi Matsumura
Kazufumi Takano
Shigenori Kanaya

概要

The crystal structure of glycerol kinase from the hyperthermophilic archaeon Thermococcus kodakaraensis (Tk-GK) in a dimeric form was determined at a resolution of 2.4 A. This is the first crystal structure of a hyperthermophilic glycerol kinase. The overall structure of the Tk-GK dimer is very similar to that of the Escherichia coli glycerol kinase (Ec-GK) dimer. However, two dimers of Ec-GK can associate into a tetramer with a twofold axis, whereas those of Tk-GK cannot. This may be the reason why Tk-GK is not inhibited by fructose 1,6-bisphosphate, because the fructose 1,6-bisphosphate binding site is produced only when a tetrameric structure is formed. Differential scanning calorimetry analyses indicate that Tk-GK is a highly thermostable protein with a melting temperature (T(m)) of 105.4 degrees C for the major transition. This value is higher than that of Ec-GK by 34.1 degrees C. Comparison of the crystal structures of Tk-GK and Ec-GK indicate that there is a marked difference in the number of ion pairs in the alpha16 helix. Four ion pairs, termed IP1-IP4, are formed in this helix in the Tk-GK structure. To examine whether these ion pairs contribute to the stabilization of Tk-GK, four Tk-GK and four Ec-GK derivatives with reciprocal mutations at the IP1-IP4 sites were constructed. The determination of their stabilities indicates that the removal of each ion pair does not affect the stability of Tk-GK significantly, whereas the mutations designed to introduce one of these ion pairs stabilize or destabilize Ec-GK considerably. These results suggest that the ion pairs in the alpha16 helix contribute to the stabilization of Tk-GK in a cooperative manner.

発表雑誌等の名称

The FEBS journal

出版者

WILEY-BLACKWELL

275

10

開始ページ

2632

終了ページ

43

発行又は発表の年月

2008-05

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

10.1111/j.1742-4658.2008.06410.x

ID:NAID(CiNiiのID)

 

ID:PMID

 

JGlobalID

 

arXiv ID

 

ORCIDのPut Code

 

DBLP ID