DeLacey Antonio L, Fernandez Victor M, Rousset Marc, Cavazza Christine, Hatchikian E Claude
Instituto de Catalisis, CSIC, Campus de Cantoblanco, 28049 Madrid, Spain.
J Biol Inorg Chem. 2003 Jan;8(1-2):129-34. doi: 10.1007/s00775-002-0397-4. Epub 2002 Sep 13.
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.
已对果糖脱硫弧菌镍铁氢化酶活性位点附近的氨基酸残基进行了定点诱变。通过傅里叶变换红外光谱对不同的突变体进行了分析，并与野生型酶进行了比较。光谱中观察到的变化证实，活性位点铁原子的氰根配体与某些相邻氨基酸残基之间的氢键稳定了蛋白质基质中的活性中心。然而，对突变体的动力学分析表明，所取代的残基在氢化酶的催化机制中均无重要作用。
