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THERMODYNAMICS OF LIGAND BINDING TO HUMAN ERYTHROCYTE

与人红细胞结合的配体的热力学

基本信息

批准号：
3438496
负责人：
CHRIS L CRANEY
金额：
$ 6.6万
依托单位：
OCCIDENTAL COLLEGE
依托单位国家：
美国
项目类别：
财政年份：
1986
资助国家：
美国
起止时间：
1986-09-01 至 1989-08-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/3438496
关键词：
NAD(H) phosphate enzyme substrate erythrocytes glucose 6 phosphatase glucose 6 phosphate dehydrogenase high performance liquid chromatography human tissue thermodynamics

项目摘要

The proposed research examines the thermodynamics of substrate binding to human erythrocyte glucose-6-phosphate dehydrogenase (G6PD), a key enzyme of the pentose phosphate pathway. The project's health implication is derived from the importance of G6PD. G6PD maintains the reducing environment of the erythrocyte, and in genetically deficient individuals, it is suspected of contributing to hemolysis. The specific aims of the proposal are to measure quantitatively the number of nicotinamide adenine dinucleotide phosphate (NADP) and glucose-6-phosphate (G6P) molecules bound to G6PD and the magnitude of the association constant for these two molecules. The project will directly measure the equilibrium quantity of radioactively labeled G6P, or NADP, bound to the enzyme. These measurements will be conducted under pH and concentration of potential regulatory molecules (e.g. ATP) conditions known to influence the enzymatic activity of G6PD. The binding data will be analyzed by direct fitting to a series of binding equations of increasing complexity using a non-linear regression algorithm. The algorithm will yield quantitative values for the number and strength of the ligand binding. Comparison between these measurements will indicate the effect of these ligands on G6PD, and analysis of the binding profiles by a linked-functions approach will provide quantitative estimates of the impact of regulatory molecules upon substrate binding.

拟议的研究检查了底物结合的热力学人红细胞葡萄糖-6-磷酸脱氢酶（G6 PD）是一种关键酶，戊糖磷酸途径。该项目的健康影响是由 G6 PD的重要性。 G6 PD维持了红细胞，并在遗传缺陷的个人，这是怀疑导致溶血该提案的具体目标是定量测定烟酰胺腺嘌呤二核苷酸的数量磷酸（NADP）和葡萄糖-6-磷酸（G6 P）分子结合到G6 PD，这两个分子的缔合常数的大小。的该项目将直接测量放射性物质的平衡量标记的G6 P，或NADP，与酶结合。这些测量将是在pH和潜在调节分子浓度下进行 (e.g. ATP）条件下，已知影响G6 PD的酶活性。结合数据将通过直接拟合一系列结合来分析。使用非线性回归的增加复杂性的方程算法该算法将产生数字的定量值，配体结合的强度。这些测量值之间的比较将表明这些配体对G6 PD的影响，并分析结合通过链接功能方法的配置文件将提供定量估计调节分子对底物结合的影响。