Role of Cysteine Residues in 4-Oxalomesaconate Hydratase from Pseudomonas ochraceae NGJ1
スポンサーリンク
概要
- 論文の詳細を見る
4-Oxalomesaconate hydratase from Pseudomonas ochraceae NGJ1 is unstable in the absence of reducing reagents such as dithiothreitol, and strongly inhibited by 5,5′-dithiobis(2-nitrobenzoic acid) (DTNB). To study the role of cysteine residues in enzyme catalysis, the eight individual cysteine residues of the enzyme were replaced with serine residues by site-directed mutagenesis. The catalytic properties and chemical modification of wild- and mutant type-enzymes by DTNB showed that (i) none of eight cysteine residues was essential for enzyme catalysis; (ii) the inhibition by DTNB was mostly due to modification of Cys-186; (iii) Cys-96 might be another residue reacting with DTNB, and its modification caused an increase in the Km-value for 4-oxalomesaconate; (iv) the other six cysteine residues were inaccessible to DTNB, but susceptible to HgCl2; and (v) only replacement of Cys-186 remarkably improved the stability of the enzyme in the absence of reducing reagent.
- 社団法人 日本農芸化学会の論文
- 2007-02-23
著者
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HAYASHI Kunihiko
Department of Basic Allied Medicine, School of Health Sciences, Faculty of Medicine, Gunma Universit
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Hayashi Kunihiko
Department Of Basic Allied Medicine School Of Health Sciences Faculty Of Medicine Gunma University
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INOUE Kazunori
Department of Electronic Science and Engineering, Kyoto University
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MARUYAMA Kiyofumi
Department of Biomolecular Science, Faculty of Engineering, Gifu University
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Inoue Kazunori
Department Of Biomolecular Science Faculty Of Engineering Gifu University
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LI Suhong
Department of Biomolecular Science, Faculty of Engineering, Gifu University
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KIMURA Maho
Department of Biomolecular Science, Faculty of Engineering, Gifu University
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TAKASHIMA Teruo
Department of Biomolecular Science, Faculty of Engineering, Gifu University
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ISHIGURO Ryou
Department of Biomolecular Science, Faculty of Engineering, Gifu University
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SUGISAKI Hiroyuki
Institute for Chemical Research, Kyoto University
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Takashima Teruo
Department Of Biomolecular Science Faculty Of Engineering Gifu University
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Maruyama Kiyofumi
Dep. Of Biomolecular Sci. Fac. Of Engineering Gifu Univ.
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Maruyama Kiyofumi
Department Of Biomolecular Science Faculty Of Engineering Gifu University
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Kimura Maho
Department Of Biomolecular Science Faculty Of Engineering Gifu University
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Ishiguro Ryou
Department Of Biomolecular Science Faculty Of Engineering Gifu University
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Hayashi Kunihiko
Department Of Biomolecular Science Faculty Of Engineering Gifu University
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Li Suhong
Department Of Biomolecular Science Faculty Of Engineering Gifu University
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Sugisaki Hiroyuki
Institute For Chemical Research Kyoto University
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Sugisaki Hiroyuki
Inst. For Chemical Res. Kyoto Univ.
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