LASER RAMAN SCATTERING FROM BIOLOGICAL MACROMOLECULES

本提案的目的是确定大分子 酶、核酸的构象和大分子动力学， 在水溶液中以更定量的方式使用膜组分，或 水分散体比以前可能使用两个经典的 和共振激光拉曼散射。 采用两种流动式冷冻淬火 和稳流技术与散射拉曼的摄像机检测 信号应提供有关瞬态结构的重要信息， 酶-底物中间体和代谢途径中的中间体 蛋白质折叠。 构象的初步研究 脱氧核糖核苷酸寡聚体显示出显著的差异， 晶体和溶液中的构象。 建议制定一个 寡核苷酸碱基序列依赖性系统研究 二级结构 两个新的光谱仪一个使用冷冻淬火 一种是旋转池技术，另一种是稳态流动系统， 已经开发，并建议使用这些工具， 研究酶-底物复合物。 新的方法来获得 二级结构的平衡构象状态和 使用经典的方法在蛋白质重折叠过程中的动力学构象状态 发展拉曼技术。 测定生物量的新方法 二级结构依赖的波动，导致 氢-氘交换也正在开发中。 由于许多健康 相关的问题与生物的正常功能密切相关。 大分子，更好地了解这些分子的动力学 应该有助于预防和一般保健。

The objectives of this proposal are to determine the macromolecular conformation and macromolecular dynamics of enzymes, nucleic acids, and membrane components in a more quantitative manner in aqueous solutions or aqueous dispersions than has previously been possible using both classical and resonance laser Raman scattering. The use of both flow freeze-quench and steady flow techniques with vidicom detection of the scattered Raman signal should give important information on the structure of transients of both enzyme-substrate intermediates and intermediates in the pathway of protein folding. Preliminary studies on conformation of deoxyribonucleotide oligomers show remarkable differences between the conformation in the crystal and in solution. It is proposed to develop a systematic study of nucleic base sequence dependence of oligonucleotide secondary structure. Two new spectrometers -- one using a freeze-quench spinning cell technique and the other using a steady-state flow system -- have been developed and it is proposed to use these instruments for studying enzyme-substrate complexes. New methods for obtaining the secondary structure of both the equilibrium conformational states and the kinetic conformational states in a protein during refolding using classical Raman technique will be developed. New methods for determining the secondary structure dependence of fluctuations leading to hydrogen-deuterium exchange are also being developed. Since many health related problems are closely linked to the proper function of biological macromolecules, a better understanding of the dynamics of these molecules should help in both preventive and general health care.