Low Temperature Trapping of Intermediates in Enzyme- Catalyzed Reactions

这项研究涉及方法开发的组合， 获取有关酶的结构信息- 底物或酶-抑制剂复合物，以及在 早期慢结合过渡态类似物抑制剂研究 蛋白酶催化和糖苷酶中间体中的事件 催化作用 调查的重点是 衰减全反射傅立叶变换 转移红外光谱法研究酶-底物 中间体，在低温下稳定，和酶- 抑制剂复合物，以及固定化酶和 冷冻酶学研究中的停流动力学。 FT-IR具有 不仅可以揭示酶-配体中形成的新键， 复合物，但也键变形和极化。 酶在众多科学领域的重要性， 医学和工业无需赘述。 最近 分子生物学和生物技术的发展已经导致 对酶学各个方面的兴趣大大增加。 一 这是确定酶细节的主要问题。 催化机理是酶底物寿命短 复合物和中间体，使其特别困难 以获取结构信息。 正常人的寿命 瞬时物种可以显著增加， 低温 开发一些方法， 促进结构数据的获取正在进行中 这里.

This research involves a combination of method development for the acquisition of structural information concerning enzyme- substrate or enzyme-inhibitor complexes, and application to the study of slow-binding transition-state analog inhibitors, early events in protease catalysis and intermediates in glycosidase catalysis. The focus of the investigation is on the application of attenuated total reflectance (ATR) Fourier transfer infrared spectroscopy to study enzyme-substrate intermediates, stabilized at low temperatures, and enzyme- inhibitor complexes, and the use of immobilized enzymes and stopped-flow kinetics in cryoenzymology studies. FT-IR has the potential to reveal not only new bonds formed in enzyme-ligand complexes, but also bond deformations and polarization. The importance of enzymes in numerous areas of science, medicine and industry needs no elaboration. Recent developments in molecular biology and biotechnology have led to a greatly increased interest in all aspects of enzymology. A major problem in determining the details of an enzyme. Catalytic mechanism is the short lifetime of enzyme - substrate complexes and intermediates, making it particularly difficult to obtain structural information. The lifetimes of normal transient species can be significantly increased at sintably low temperatures. The development of some methods which should facilitate the acquisition of structural data are underway here.