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Gene Synthesis, Expression, and Mutagenesis of Zucchini Mavicyanin: The Fourth Ligand of Blue Copper Center Is Gln.

フォーマット:
論文
責任表示:
片岡, 邦重 ; Kataoka, Kunishige ; Nakai, Masami ; Yamaguchi, Kazuya ; Suzuki, Shinnichiro
言語:
英語
出版情報:
Elsevier, 1998-01-01
著者名:
掲載情報:
Biochemical and Biophysical Research Communications
ISSN:
0006-291x  CiNii Research  Webcat Plus  JAIRO
巻:
250
通号:
2
開始ページ:
409
終了ページ:
413
バージョン:
author
概要:
The gene coding for the 109-amino-acid, non-glycosylated form of mavicyanin was synthesized and expressed inEscherichia coli.The recombinant protein refolded fromE. coliinclusion bodies was purified and characterized. Its spectroscopic properties are fully identical to those of mavicyanin isolated from zucchini, even in the absence of its carbohydrate moiety. The blue cooper center of mavicyanin strongly binds three ligands (2His and Cys) as well as many blue copper proteins. To disclose the fourth ligand of mavicyanin, Met was substituted for Gln95 by site-directed mutagenesis. The replacement changes from a rhombic EPR signal to an axial one and exhibits the quite similar absorption and CD spectra to those of plastocyanin. The midpoint potential of Gln95→Met mavicyanin shows the positive shift of 187 mV compared with the recombinant protein, being close to the values of plastocyanins. The differences of the spectroscopic and electrochemical properties between mavicyanin and its mutant demonstrate that the fourth ligand of mavicyanin is Gln95. 続きを見る
URL:
http://hdl.handle.net/2297/1733
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