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系统，用于进行急冷流动。 反应动力学在反应机理研究中的应用 精子网、ATPase和其他酶系统不能 其次是光学手段。时间分辨率优于1.5毫秒的 到目前为止，通过2-4反应的酸猝灭实现了 以乙酸二硝基苯酯为试验反应。每种试剂0.25毫升 每个数据点需要2.5毫升，而其他仪器则为2.5毫升。 计划将其用于零下40摄氏度的低温淬火 即将到来的一年。该驱动器在我们的零压降热力系统中的使用 停流系统是去年报告的。压力、速度和 在一系列粘度上正在进行的热测量提供了 支持球体混合器理论所需的信息，即 混合主要是线动量变为角动量的结果。 势头。到目前为止的结果表明，对于粘度为1的 使压差小于10磅/平方英寸就足以 以3M/秒的速度产生比99%更好的混合，仅有 正负2M摄氏度。此外，还将增加光纤传感器 在接下来的一年里，热-光同时停止流动 实验是可以做的。