SURFACE-ENHANCED RAMAN STUDIES OF BIOMOLECULES

本研究的长期目标是发展Sers/SERRS作为一种通用的 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拉曼光谱具有相当大的潜力， 基础研究和生物分析应用。期间 该工作程序的初始阶段用于获得Sers/SERRS光谱， 开发了各种生物分子， 结构该技术对结构/功能的适用性 水溶性和膜结合分析（光合和间隙 连接）蛋白质，核酸和模型系统， 演示。在拟议的供资期间， Sers技术与电化学方法、可见光、UV-RR和 时间分辨拉曼光谱，将用于研究的问题， 这是一项重要的工作，并开发了一种新的分析技术。 要处理的两个具体目标包括： (1)SERRS、共振和UV共振拉曼光谱，以及 伏安法测量将适用于各种细胞色素 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。 的 SERRS与电化学方法的组合将用于以下 在电子转移过程中发生的血红素结构变化 过程UV-RR散射将提供芳香族氨基酸的探针 不同突变体中的残基。时间分辨R&an光谱将是 用于测量电子转移之间的动力学 细胞色素和它们的天然氧化还原伙伴在溶液中， 电极表面使用SERRS技术。化学修饰 细胞色素c和共价连接的细胞色素c/质体蓝素复合物 也将被研究。 (2)SERRS-连接免疫分析（SLI）是一种新的、高灵敏度的 和高度特异性的方法。 一种通用的SLI手术，可用于不同的临床和 将在拟议供资期间进行环境分析 期两种类型的模型系统将被检查，以验证 这种方法：以百草枯为例， 铁蛋白作为临床大分子重要性和实例 应用程序.目标是达到阿托摩尔浓度的检测极限 范围

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.