VIBRATIONAL SPECTROSCOPY OF REDOX METALLOPROTEINS

这项建议旨在应用现代振动光谱 探针（主要共振拉曼（RR）、红外（IR）、时间- 解析激光光谱学和相关技术）来选择 氧化还原金属蛋白的化学和生物物理学问题。 其目的是了解基本因素， 影响这些基本的活性位点的功能， 生物分子 我们关心的是这些蛋白质如何影响 它们的关键代谢功能是容易的电子转移， 小分子（例如，02)激活，必然的现象，如 分子开关和电子转移的热力学调节， 和耦合功能如氧化还原相关的质子泵。 我们将 集中在“蓝色”铜蛋白和细胞色素c氧化酶 （CcO）。 选择这些系统是因为其内在的 利益和所涉问题的普遍性。 具体 拟开展的项目包括以下内容。 1. 经常资源研究旨在确定 振动观测量、结构函数和动力学 蓝铜活性中心的位置 2. CcO的基本RR和IR表征，包括 详细的RR激发曲线和同位素研究。 3. CcO动力学的时间分辨振动研究， 表征连接和电子转移的目的 酶的功能反应性的瞬变。 4. RR研究旨在阐明质子易位 CCO的功能。 5. Rieske型铁硫化合物的振动特性 氮蛋白 6. 新发现的细胞色素的振动1表征 嗜热栖热菌的beta alpha 3 7. 表征合成复合物，其可以模拟 CcO的Fe-Cu偶联细胞色素α 3-CuB位点 8. 极压对光谱影响的研究 和外部金属的长程电子转移动力学， 标记的和金属取代的蛋白质。

This proposal seeks to apply modern vibrational spectroscopic probes (Principally resonance Raman (RR), infrared (IR), time- resolved laser spectroscopies, and related techniques) to selected problems in the chemistry and biophysics of redox metalloproteins. The objective is to understand the fundamental factors which influence the function of the active sites of these essential biomolecules. We are concerned with how these proteins effect their critical metabolic functions of facile electron transfer and small molecule (e.g., 02) activation, corollary phenomena such as molecular switching and thermodynamic tuning of electron transfer, and coupled functions such as redox-linked proton pumping. We will concentrate on the "blue" copper proteins and cytochrome c oxidase (CcO). These systems are chosen because of their intrinsic interest and the generality of the issues involved. Specific projects to be undertaken include the following. 1. RR studies aimed at establishing specific relationships between the vibrational observables and the structure function and dynamics of the blue copper active site. 2. Fundamental RR and IR characterization of CcO, including detailed RR excitation profiles and isotope studies. 3. Time-resolved vibrational studies of CcO dynamics, with the objective of characterizing the ligation and electron transfer transients in the functional reactivity of the enzyme. 4. RR studies aimed at elucidating the proton translocation function of CcO. 5. Vibrational characterization of Rieske-type iron-sulfur- nitrogen proteins. 6. Vibration 1 characterization of the newly discovered cytochrome beta alpha 3 from Thermus thermophilus. 7. Characterization of synthetic complexes which may mimic the coupled Fe-Cu cytochrome alpha 3-CuB site of CcO 8. Study of the effects of extreme pressure upon the spectroscopy and long-range electron transfer dynamics of externally metal- labelled and metal-substituted proteins.