DIRECT MEASUREMENT OF ENZYMATIC ACTIVITIES BY MICROCALORIMETRY: HIV PROTEASE

通过微量热法直接测量酶活性：HIV 蛋白酶

• 批准号: 6122012
• 负责人: MATTHEW J TODD
• 金额: --
• 依托单位: JOHNS HOPKINS UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 1997
• 资助国家: 美国
• 起止时间: 1997-08-05 至 1998-08-04
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6122012
• 关键词: AIDS; biological products; biomaterials; biomedical equipment development; biomedical resource; communicable diseases; education; enzymes; human tissue; immunology; lymphatic system; model design /development; proteins; technology /technique development; virus

A widely-applicable method for measuring enzymatic activity using micro-calorimetry is being developed. Chemical reactions are driven by a decrease in free energy, which is the sum of enthalpic and entropic contributions. The enthalpic contribution is characterized by heat. Using isothermal titration calorimetry, we measure the heat evolved during a reaction and express the results as reaction rates. Whereas classical enzyme assays calculate rate as a derivative of product appearance (or substrate depletion) with time, biocalorimetric assays measure reaction rates directly. Isothermal calorimetric analysis is a non-destructive (non-radioactive) technique that 1) is applicable to most biological reactions, 2) eliminates the requirement for coupling enzymes, 3) is useful in spectrophotometrically opaque solutions, and 4) permits rapid activity analysis using physiological substrates. We present data assaying enzymes from; E. Coli chaperonin GroEL, HIV-1 protease, F. Heparinium heparinase, and H. Pylorii urease.

一种广泛适用的测定酶活性的方法， 正在开发微量热法。 化学反应是由 通过自由能的减少，自由能是自由能和自由能的总和， 熵的贡献 的贡献的特点是 通过加热。 使用等温滴定量热法， 在反应过程中演化，并将结果表示为反应速率。 而经典的酶测定将速率计算为 产物外观（或底物消耗）随时间的变化，生物量热法 测定直接测量反应速率。 等温量热 分析是一种非破坏性（非放射性）技术，1） 适用于大多数生物反应，2）消除了要求 对于偶联酶，3）可用于光学不透明的 4）允许使用生理学方法进行快速活性分析 印刷受体. 我们目前的数据分析酶; E。大肠杆菌伴侣蛋白 GroEL、HIV-1蛋白酶、F.肝素肝素酶， 和H.幽门螺旋菌尿素酶。