PROKARYOTIC PHOSPHORIBULOKINASE

原核磷酸核糖激酶

• 批准号: 6307853
• 负责人: HENRY M MIZIORKO
• 金额: $ 1.13万
• 依托单位: MEDICAL COLLEGE OF WISCONSIN
• 依托单位国家: 美国
• 财政年份: 2000
• 资助国家: 美国
• 起止时间: 2000-03-01 至 2001-02-28
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6307853
• 关键词: bioengineering /biomedical engineering; biological products; biomedical resource; enzymes; proteins

Mutagenic studies that implicate specific amino acids in reaction chemistry or 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 appropriate spin-labeled substrate analogs. Measurement of spin-probe affinity, binding stoichiometry, and correlation time using D42A and D169A 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. A spin-labeled ATP analog (ATPSAP) is being productively used to validate the integrity of the engineered variants D42A and D169A of prokaryotic phosphoribulokinase which catalyzes a key step in carbon dioxide assimilation in Calvin's reductive pentose phosphate cycle.

涉及反应中特定氨基酸的诱变研究 化学或底物结合只有在以下情况下才能完全解释： 变异蛋白的结构完整性已得到验证。 通过识别适当的信息可以促进这种验证 自旋标记底物类似物。 自旋探针亲和力的测量， 使用 D42A 和 D169A 的结合化学计量和相关时间 突变体并将这些参数与测量的参数进行比较 野生型蛋白质提供了优雅、严谨且高效的方法 验证结构完整性的解状态方法 工程蛋白质。 自旋标记 ATP 类似物 (ATPSAP) 正在开发中 有效地用于验证工程变体的完整性 原核磷酸核糖激酶的 D42A 和 D169A 催化 卡尔文还原戊糖中二氧化碳同化的关键步骤 磷酸盐循环。