SURFACE-ENHANCED RAMAN STUDIES OF BIOMOLECULES

这项研究的长期目标是将SERS/SERS发展为一般的 研究生物系统的技术。巨大的敏感性 由增强的拉曼效应提供的具有相当大的潜力 基础研究和生物分析应用。在.期间 获取SERS/SERRS谱的工作程序的初始阶段 各种生物分子被开发出来，以保护原住民 结构。这项技术在结构/功能方面的适用性 水溶性和膜结合态(光合作用和GAP)的分析 连接)蛋白质、核酸和模型系统的成功 演示了。在拟议的供资期间，以前制定的 SERS技术与电化学方法相结合，可见、UV-RR和 时间分辨拉曼光谱，将用来研究一个问题 根本利益，并开发新的分析技术。 需要解决的两个具体目标包括： (1)SERRS、共振和UV共振拉曼光谱，以及 伏安法将应用于多种细胞色素。 C和细胞色素c3突变体，以表征蛋白质/血红素 相互作用及其对电子转移性质的影响。这个 将使用SerRS和电化学方法相结合的方法进行后续研究 血红素在电子转移过程中发生的结构变化 进程。UV-RR散射将为芳香族氨基酸提供探针 不同突变体中的残留物。时间分辨R&A光谱学将是 用来测量电子在两个原子之间转移的动力学 细胞色素及其天然氧化还原伙伴在溶液和 电极表面采用SERRS技术。化学改性 细胞色素c和共价连接的细胞色素c/胞质蓝蛋白络合物 也将被研究。 (2)提出了一种新的、高灵敏度的血清免疫分析(SLI)方法 以及高度特异的检测生物重要分子的方法。 一种通用的SLI程序，可用于不同的临床和 环境分析将在拟议的资金期间进行。 句号。将检查两种类型的模型系统以验证 这种方法：以百草枯为例的小分子与环境 大分子铁蛋白在临床中的重要性和应用 申请。目标是在大气层中达到检测极限 射程。

The long term goal of this research is to develop SERS/SERRS as a general technique for the study of biological systems. The enormous sensitivity provided by the enhanced Raman effect has considerable potential for fundamental studies and also for bioanalytical applications. During the initial phase of this work procedures for obtaining SERS/SERRS spectra for a variety of biological molecules were developed which preserve the native structure. The applicability of this technique for structure/functional analysis of water soluble and membrane bound (photosynthetic and gap junction) proteins, nucleic acids and model systems was successfully demonstrated. During the proposed funding period the previously developed SERS technique, together with electrochemical methods, visible, Uv-RR and time resolved Raman spectroscopy, will be used to study a problem of fundamental interest and to develop a new analytical technique. The two specific aims to be addressed include the following: (1) SERRS, resonance and UV resonance Raman spectroscopy, together with voltammetric measurements will be applied to a wide variety of cytochrome c and cytochrome c3 mutants in order to characterize the protein/heme interactions and their effect on the electron transfer properties. The combination of SERRS with electrochemical methods will be used to follow the structural changes in the heme which occur during electron transfer process. UV-RR scattering will provide a probe of the aromatic amino acid residues in different mutants. Time resolved R&an spectroscopy will be utilized to measure the kinetics of electron transfer between the cytochromes and their natural redox partners in solution and at the electrode surface using the SERRS technique. Chemically modified cytochrome c and covalently linked cytochrome c/plastocyanin complexes will be studied as well. (2) SERRS-Linked Immunoassay (SLI) is proposed as a new, highly sensitive and highly specific method for detecting biologically important molecules. A universal SLI procedure which can be used for different clinical and environmental analyses will be developed during the proposed funding period. Two types of model systems will be examined in order to validate this method: paraquat as an example of small molecules with environmental importance and ferritin as an example of a large molecule for clinical application. The goal will be to reach detection limits in the attomolar range.