DEVELOPMENT OF BIOPHYSICAL METHODS FOR STUDY OF BIO-MACROMOLECULAR REACTIONS

研究生物大分子反应的生物物理方法的发展

• 批准号: 4694533
• 负责人: R L BERGER
• 金额: --
• 依托单位: NATIONAL HEART, LUNG, AND BLOOD INSTITUTE
• 依托单位国家: 美国
• 资助国家: 美国
• 起止时间: 至
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/4694533
• 关键词: acidity /alkalinity; adenosinetriphosphatase; cell membrane; chemical reaction; enzyme substrate; fiber optics; flash photolysis; hemoglobin; lactate dehydrogenases; lasers; muscle contraction; sarcoplasmic reticulum; sickle cell anemia; stop flow technique; thermodynamics; viscosity

A new inertial drive flow system has been developed for doing quench flow reaction kinetics in the investigation of the mechanism of reaction of Sacroplasmic Reticulum, ATPase, and other enzyme systems which cannot be followed by optical means. A time resolution of better than 1.5 ms has been achieved to date using the acid quenching of the reaction of 2-4 dinitrophenyl acetate as the test reaction. 0.25ml of each reagent are required per data point in contrast to 2.5 ml for other instruments. Extension of its use for cryoquenching at -40 degrees C is planned for the coming year. The use of this drive in our zero pressure drop thermal stopped flow system was reported last year. Pressure, velocity, and thermal measurements underway over a series of viscosities have provided information needed for the support of the ball mixer theory, i.e. that mixing is mainly the result of changing linear momentum to angular momentum. The results to date demonstrate that for a viscosity of one centipoise a pressure differential of less that 10 psi is sufficient to produce better than 99% mixing at 3 M/sec with thermal fluctuations of only plus/minus 2m degrees C. In addition optical fiber sensors will be added in the coming year so that simultaneous thermal-optical stopped flow experiments may be done.

开发了一种新的惯性驱动流动系统用于淬火流动 反应动力学在反应机理研究中的应用 肌浆网、ATP 酶和其他不能被 其次是光学手段。 时间分辨率优于 1.5 ms 迄今为止，使用酸猝灭2-4的反应已实现 乙酸二硝基苯酯作为测试反应。 每种试剂0.25ml 每个数据点需要 2.5 毫升，而其他仪器则需要 2.5 毫升。 计划将其用于 -40 摄氏度的低温淬火 来年。 在我们的零压降热工中使用该驱动器 去年报道了停流系统。 压力、速度和 对一系列粘度进行的热测量提供了 支持球混合器理论所需的信息，即 混合主要是线动量转变为角动量的结果 势头。 迄今为止的结果表明，对于粘度为 1 厘泊 小于 10 psi 的压差就足以 以 3 M/秒的速度产生优于 99% 的混合，热波动仅为 正/负 2m 摄氏度。此外还将添加光纤传感器 在来年，以便同时热光停止流动 可以进行实验。