Structure-Function Studies of Alcohol Dehydrogenases

醇脱氢酶的结构功能研究

• 批准号: 9118657
• 负责人: Bryce Plapp
• 金额: $ 26.4万
• 依托单位: University of Iowa
• 依托单位国家: 美国
• 项目类别: Continuing Grant
• 财政年份: 1992
• 资助国家: 美国
• 起止时间: 1992-06-01 至 1995-11-30
• 项目状态: 已结题
• 来源: https://www.nsf.gov/awardsearch/showAward?AWD_ID=9118657&HistoricalAwards=false
• 关键词: Structure; Function; Studies; Alcohol; Dehydrogenases

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.

将使用定点诱变来取代氨基酸 这些酶中的残基，以及稳态和瞬态 动力学、X射线晶体学和其他方法将用于 研究这些变量。 质子中继器中的氨基酸残基 系统，和环境的催化锌，将进行研究。 蛋白质的灵活性和氢隧道将在 肝酶 “外来”结构要素，包括 结构锌环和其他环，将被删除。 的 将探索低聚结构的起源。 马肝酶的荧光特性将是 通过X射线晶体学测定。 %%% 研究了马肝和酵母中的乙醇脱氢酶 广泛地。 拟议研究的目标是回答 关于催化机理的几个悬而未决的问题， 酶，底物和辅酶特异性的进化， 以及三维结构在酶动力学中的作用 和催化作用。 这项研究应该增加我们对 生物催化