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タイトル: Four-electron reduction of dioxygen by a multicopper oxidase, CueO, and roles of Asp112 and Glu506 located adjacent to the trinuclear copper center
著者: Kataoka, Kunishige link image link image
Sugiyama, Ryosuke
Hirota, Shun
Inoue, Megumi
Urata, Kanae
Minagawa, Yoichi
Seo, Daisuke link image link image
Sakurai, Takeshi link image link image
片岡, 邦重
瀨尾, 悌介
櫻井, 武
発行日: 2009年 5月22日
出版社(者): American Society for Biochemistry and Molecular Biology
雑誌名: Journal of Biological Chemistry
ISSN: 0021-9258
巻: 284
号: 21
開始ページ: 14405
終了ページ: 14413
抄録: The mechanism of the four-electron reduction of dioxygen by a multicopper oxidase, CueO, was studied based on reactions of single and double mutants with Cys500, a type I copper ligand, and the noncoordinating Asp112 and Glu506, which form hydrogen bonds with the trinuclear copper center directly and indirectly via a water molecule. The reaction of C500S containing a vacant type I copper center produced intermediate I in an EPR-silent peroxide-bound form. The formation of intermediate I from C500S/D112N was restricted due to a reduction in the affinity of the trinuclear copper center for dioxygen. The state of intermediate I was realized to be the resting form of C500S/E506Q and C500S of the truncated mutant Δα5-7CueO, in which the 50 amino acids covering the substrate-binding site were removed. Reactions of the recombinant CueO and E506Q afforded intermediate II, a fully oxidized form different from the resting one, with a very broad EPR signal, g < 2, detectable only at cryogenic temperatures and unsaturated with high power microwaves. The lifetime of intermediate II was prolonged by the mutation at Glu506 involved in the donation of protons. The structure of intermediates I and II and the mechanism of the four-electron reduction of dioxygen driven by Asp112 and Glu506 are discussed. © 2009 by The American Society for Biochemistry and Molecular Biology, Inc.
DOI: 10.1074/jbc.M808468200
URI: http://hdl.handle.net/2297/18720
資料種別: Journal Article
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このアイテムを引用あるいはリンクする場合は次の識別子を使用してください。 http://hdl.handle.net/2297/18720



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