Sequence motif-specific assignment of two [2Fe-2S] clusters in rat xanthine oxidoreductase studied by site-directed mutagenesis

被引：40
|
作者：
Iwasaki, T
Okamoto, K
Nishino, T [1 ]
Mizushima, J
Hori, H
Nishino, T [1 ]
机构：
[1] Nippon Med Sch, Dept Biochem & Mol Biol, Tokyo 1138602, Japan
[2] Yokohama City Univ, Sch Med, Dept Biochem, Yokohama, Kanagawa 2360004, Japan
来源：
JOURNAL OF BIOCHEMISTRY | 2000年 / 127卷 / 05期
关键词：
electron paramagnetic resonance spectroscopy; iron-sulfur cluster; site-directed mutagenesis; xanthine dehydrogenase; xanthine oxidase;
D O I：
10.1093/oxfordjournals.jbchem.a022669
中图分类号：
Q5 [生物化学]; Q7 [分子生物学];
学科分类号：
071010 ; 081704 ;
摘要：
The sequence motif-specific assignment of the two distinct [2Fe-2S] clusters in rat xanthine oxidoreductase (XOR) was unequivocally established by site-directed mutagenesis of recombinant enzymes expressed in a baculovirus-insect cell system and electron paramagnetic resonance (EPR) spectroscopy. The conserved cysteine residues, including Cys-115, in the unusual C-terminal -Cys-Xaa(2)-Cys-//-Cys-Xaa(1)-Cys- motif serve as ligands to the Fe/S I center, which is probably located in close proximity to the Mopterin center. Other conserved cysteine residues, including Cys-43 and Cys-51, in the N-terminal plant ferredoxin-like motif serve as ligands to the Fe/S II center, which is distantly located from the Mo-pterin center. The present sequence motif-specific assignment of the Fe/S I and II centers is discussed in the light of the structural features of XOR.
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收藏
页码：771 / 778
页数：8
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