VIBRATIONAL SPECTROSCOPY OF REDOX METALLOPROTEINS

这项提议试图应用现代振动光谱 探头(主要是共振拉曼(RR)、红外(IR)、时间- 解析的激光光谱分析和相关技术)以选定 氧化还原金属蛋白的化学和生物物理问题。 目标是了解以下基本因素： 影响这些必需的活性部位的功能 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轻便的电子传递和 小分子(例如，02)活化，由此产生的现象，如 电子转移的分子开关和热力学调节， 以及氧化还原连接的质子泵浦等耦合功能。我们会 关注“蓝色”铜蛋白和细胞色素c氧化酶 (CCO)。选择这些系统是因为它们的内在 兴趣和所涉问题的一般性。特定的 将开展的项目包括以下项目。 1.旨在建立特定关系的RR研究 振动能观与结构函数和动力学 蓝铜活跃点的。 2.CcO的基本RR和IR表征，包括 详细的RR激发谱和同位素研究。 3.CcO动力学的时间分辨振动研究 目的研究连接和电子转移的性质 酶的功能反应性的瞬变。 4.旨在阐明质子转移的RR研究 Cco的功能。 5.Rieske型铁硫化合物的振动特性 含氮蛋白质。 6.新发现的细胞色素的振动1特性 嗜热嗜热菌的β-3。 7.可模拟的合成络合物的表征 CcO的Fe-Cu细胞色素α3-CuB偶联位置 8.极端压力对光谱影响的研究 以及外部金属的远程电子转移动力学- 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.