Spectroscopic and kinetic characterization of active site mutants of Desulfovibrio fructosovorans Ni-Fe hydrogenase

被引：51
作者：
DeLacey, AL
Fernandez, VM
Rousset, M
Cavazza, C
Hatchikian, EC
机构：
[1] CSIC, Inst Catalisis, Madrid 28049, Spain
[2] CNRS, Bioenerget & Ingn Prot, Inst Biol Struct & Microbiol, F-13402 Marseille 20, France
来源：
JOURNAL OF BIOLOGICAL INORGANIC CHEMISTRY | 2003年 / 8卷 / 1-2期
关键词：
metalloprotein; site-directed mutagenesis; FTIR; spectroelectrochemistry; hydrogen;
D O I：
10.1007/s00775-002-0397-4
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：
Site-directed mutagenesis of amino acid residues proximate to the active site of the Ni-Fe hydrogenase of Desulfovibrio fructosovorans has been done. The different mutants have been analyzed by FTIR spectroscopy and compared with wild type enzyme. The changes observed in the spectra confirm that hydrogen bonds between the CN- ligands of the active site's Fe atom and certain neighbor amino acid residues stabilize the active center within the protein matrix. However, kinetic analysis of the mutants indicates that none of the replaced residues have an important role in the catalytic mechanism of the hydrogenase.
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收藏
页码：129 / 134
页数：6
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