LASER RAMAN SCATTERING FROM BIOLOGICAL MACROMOLECULES

这项建议的目标是确定大分子 酶、核酸和蛋白质的构象和大分子动力学 以更定量的方式在水溶液或 水性分散体比以前使用经典的 和共振激光拉曼散射。两种流动冷冻急冷法的使用 以及Vidicom检测散射拉曼的稳流技术 信号应提供有关瞬变结构的重要信息 酶-底物途径中的中间体和中间体 蛋白质折叠。核糖核酸分子构象的初步研究 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。有人建议开发一种 寡核苷酸碱基序列依赖性的系统研究 二级结构。两台新的光谱仪--其中一台使用冷冻猝灭 旋转细胞技术和另一种使用稳态流动系统的技术-- 已经开发出来，并建议将这些仪器用于 研究酶-底物复合体。获取数据的新方法 平衡构象状态和平衡构象的二级结构 用经典方法研究蛋白质重折叠过程中的动力学构象状态 拉曼技术将会得到发展。一种新的测定方法 波动对二级结构的依赖导致 氢-氚交换也在发展中。因为许多人的健康 相关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 在预防和一般保健方面都应该有所帮助。

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.