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ID 48773
本文ファイル
著者
Oikawa, Kenta
Nakamura, Shota
Sonoyama, Takafumi
Ohshima, Atsushi
Kobayashi, Yuji
Takayama, Shin-ichi J.
Yamamoto, Yasuhiko
Uchiyama, Susumu
Hasegawa, Jun
Sambongi, Yoshihiro 大学院生物圏科学研究科 広大研究者総覧
抄録(英)
Five amino acid residues responsible for extreme stability have been identified in cytochrome c552 (HT c552) from a thermophilic bacterium, Hydrogenobacter thermophilus. The five residues, which are spatially distributed in three regions of HT c552, were replaced with the corresponding residues in the homologous but less stable cytochrome c551 (PA c551) from Pseudomonas aeruginosa. The quintuple HT c552 variant (A7F/M13V/Y34F/Y43E/I78V) showed the same stability against guanidine hydrochloride denaturation as that of PA c551, suggesting that the five residues in HT c552 necessarily and sufficiently contribute to the overall stability. In the three HT c552 variants carrying mutations in each of the three regions, the Y34F/Y43E mutations resulted in the greatest destabilization, by –13.3 kJ mol–1, followed by A7F/M13V (–3.3 kJ mol–1) and then I78V (–1.5 kJ mol–1). The order of destabilization in HT c552 was the same as that of stabilization in PA c551 with reverse mutations such as F34Y/E43Y, F7A/V13M, and V78I (13.4, 10.3, and 0.3 kJ mol–1, respectively). The results of guanidine hydrochloride denaturation were consistent with those of thermal denaturation for the same variants. The present study established a method for reciprocal mutation analysis. The effects of side-chain contacts were experimentally evaluated by swapping the residues between the two homologous proteins that differ in stability. A comparative study of the two proteins was a useful tool for assessing the amino acid contribution to the overall stability.
内容記述
This work was supported in part by grants from Hiroshima University, the Noda Institute for Scientific Research, and the Japanese Ministry of Education, Science and Culture (grants-in-aid for Scientific Research on Priority Areas).
掲載誌名
Journal of Biological Chemistry
280巻
7号
開始ページ
5527
終了ページ
5532
出版年月日
2005-02-18
出版者
The American Society for Biochemistry and Molecular Biology, Inc.
ISSN
0021-9258
1083-351X
出版者DOI
言語
英語
NII資源タイプ
学術雑誌論文
広大資料タイプ
学術雑誌論文
DCMIタイプ
text
フォーマット
application/pdf
著者版フラグ
publisher
権利情報
This research was originally published in the Journal of Biological Chemistry. Kenta Oikawa, Shota Nakamura, Takafumi Sonoyama, Atsushi Ohshima, Yuji Kobayashi, Shin-ichi J. Takayama, Yasuhiko Yamamoto, Susumu Uchiyama, Jun Hasegawa, and Yoshihiro Sambongi. Five Amino Acid Residues Responsible for the High Stability of Hydrogenobacter thermophilus Cytochrome c552. J Biol Chem. 2005; 280:5527-5532. © 2005 the American Society for Biochemistry and Molecular Biology, Inc.
関連情報URL
部局名
生物圏科学研究科