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

Effect of the disease-causing mutations identified in human ribonuclease (RNase) H2 on the activities and stabilities of yeast RNase H2 and archaeal RNase HII.

単著・共著の別

 

著者

Muhammad S Rohman
Yuichi Koga
Kazufumi Takano
Hyongi Chon
Robert J Crouch
Shigenori Kanaya

概要

Eukaryotic ribonuclease (RNase) H2 consists of one catalytic and two accessory subunits. Several single mutations in any one of these subunits of human RNase H2 cause Aicardi-Goutières syndrome. To examine whether these mutations affect the complex stability and activity of RNase H2, three mutant proteins of His-tagged Saccharomyces cerevisiae RNase H2 (Sc-RNase H2*) were constructed. Sc-G42S*, Sc-L52R*, and Sc-K46W* contain single mutations in Sc-Rnh2Ap*, Sc-Rnh2Bp*, and Sc-Rnh2Cp*, respectively. The genes encoding the three subunits were coexpressed in Escherichia coli, and Sc-RNase H2* and its derivatives were purified in a heterotrimeric form. All of these mutant proteins exhibited enzymatic activity. However, only the enzymatic activity of Sc-G42S* was greatly reduced compared to that of the wild-type protein. Gly42 is conserved as Gly10 in Thermococcus kodakareansis RNase HII. To analyze the role of this residue, four mutant proteins, Tk-G10S, Tk-G10A, Tk-G10L, and Tk-G10P, were constructed. All mutant proteins were less stable than the wild-type protein by 2.9-7.6 degrees C in T(m). A comparison of their enzymatic activities, substrate binding affinities, and CD spectra suggests that the introduction of a bulky side chain into this position induces a local conformational change, which is unfavorable for both activity and substrate binding. These results indicate that Gly10 is required to make the protein fully active and stable.

発表雑誌等の名称

The FEBS journal

出版者

WILEY-BLACKWELL

275

19

開始ページ

4836

終了ページ

49

発行又は発表の年月

2008-10

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.1742-4658.2008.06622.x

ID:NAID(CiNiiのID)

 

ID:PMID

 

JGlobalID

 

arXiv ID

 

ORCIDのPut Code

 

DBLP ID