SPECTROSCOPIC STUDIES OF ENZYME SUBSTRATE COMPLEXES

酶底物复合物的光谱研究

• 批准号: 6298208
• 负责人: GEORGE H REED
• 金额: $ 0.75万
• 依托单位: UNIVERSITY OF WISCONSIN-MADISON
• 依托单位国家: 美国
• 财政年份: 1999
• 资助国家: 美国
• 起止时间: 1999-03-01 至 2000-02-29
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6298208
• 关键词: biological products; biomaterials; biomedical resource; enzymes; nuclear magnetic resonance spectroscopy

The structures of complexes of yeast enolase and rabbit muscle pyruvate kinase have been solved to high resolution by X-ray crystallography. These structures reveal the identities of active-site residues in these two enzymes. Site-directed mutagenesis has been performed to clarify the roles of the active-site residues in catalysis. Residues believed to function as the acid/base catalysts for these enzymes are lysine 345 for yeast enolase and either arginine 72 or threonine 327 for pyruvate kinase. Both enzymes catalyze alternate reactions. NMR characterization of the products of these alternate reactions will be carried to help corroborate our hypotheses. NMR identification of the stereospecificity of reactions catalyzed by these enzymes and mutants will also be employed.

酵母烯醇酶和兔肌肉的复合物结构 丙酮酸激酶已通过X射线求解至高分辨率 晶体学。 这些结构揭示了 这两个酶中的活性位点残基。 定向诱变 已经进行了以阐明活性位点残基在中的作用 催化。 残基被认为充当酸/碱催化剂 对于这些酶，酵母烯醇酶是赖氨酸345 丙酮酸激酶72或苏氨酸327。 两种酶都催化 替代反应。 这些产品的NMR表征 替代反应将被携带以帮助证实我们的 假设。 NMR识别反应的立体特异性 这些酶和突变体也将被催化。