EUKARYOTIC HMG COA LYASE

真核 HMG COA 裂解酶

• 批准号: 6279864
• 负责人: HENRY M MIZIORKO
• 金额: $ 0.21万
• 依托单位: MEDICAL COLLEGE OF WISCONSIN
• 依托单位国家: 美国
• 财政年份: 1998
• 资助国家: 美国
• 起止时间: 1998-04-01 至 1999-02-28
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6279864
• 关键词: bioengineering /biomedical engineering; biological products; biomedical resource; enzymes; respiratory system

Mutagenic studies that implicate specific amino acids in reaction chemistry of in substrate binding are fully interpretable only when the structural integrity of the variant protein has been validated. This validation is facilitated by identification of spin-labeled substrate analogs. Measurement of spin-probe affinity and binding stoichiometry, using a H235A and H235D mutants and comparison of these parameters with those measured for wild-type protein affords an elegant, rigorous, yet efficient solution-state method for validating the structural integrity of engineered proteins. The paramagnetic cation Mn++ is being productively used to validate the integrity of engineered variants of human HMG-CoA lyase, which catalyzed a key step in leucine catabolism. Studies were conducted using X-band EPR spectroscopy.

暗示特定氨基酸的诱变研究 仅在 变体蛋白的结构完整性已得到验证。 通过识别自旋标记，可以促进此验证 底物类似物。 测量自旋探针亲和力和结合 化学计量法，使用H235A和H235D突变体并进行比较 用野生型蛋白测量的参数提供了 优雅，严格但有效的解决方案方法用于验证 工程蛋白的结构完整性。 顺磁性 阳离子Mn ++正在有效地用于验证 人类HMG-COA裂解酶的工程变体，它催化了关键步骤 在亮氨酸分解代谢中。 使用X波段EPR进行了研究 光谱法。