Structure-Function Studies of Alcohol Dehydrogenases

定点突变将被用来替代这些酶中的氨基酸残基，稳态和瞬时动力学、X射线结晶学和其他方法将被用来研究这些变异。将研究质子接力系统中的氨基酸残基，以及催化锌的环境。蛋白质的灵活性和氢隧道将在肝酶中进行研究。“无关的”结构元素，包括结构锌环和其他环，将被删除。我们将探索低聚结构的起源。马肝酶的荧光性质将通过X射线结晶学来确定。从马肝和酵母中提取的%乙醇脱氢酶已被广泛研究。这项研究的目的是回答关于酶的催化机制、底物和辅酶专一性的演变以及三维结构在酶动力学和催化中的作用等几个突出问题。这项研究应该会增加我们对生物催化的理解。

Site-directed mutagenesis will be used to substitute amino acid residues in these enzymes, and steady-state and transient kinetics, x-ray crystallography, and other methods will be used to study these variants. Amino acid residues in the proton relay system, and the environment of the catalytic zinc, will be studied. Protein flexibility and hydrogen tunneling will be studied in the liver enzyme. "Extraneous" structural elements, including the structural zinc loop and other loops, will be deleted. The origins of the oligomeric structures will be explored. Fluorescence properties of the horse liver enzyme will be determined by x-ray crystallography. %%% Alcohol dehydrogenases from horse liver and yeast have been studied extensively. The objectives of the proposed research are to answer several outstanding questions about the catalytic mechanism of the enzyme, the evolution of specificity for substrates and coenzymes, and the role of the three dimensional structure in enzyme dynamics and catalysis. This research should increase our understanding of biocatalysis.