Spectroscopy and Function of Redox Metalloproteins

描述(由申请人提供)： 这个项目的总体目标是主要通过光谱和动力学的观察，帮助理解氧化还原金属蛋白的功能和结构-功能关系。我们的方法强调振动(包括时间分辨)光谱，它对构象、氧化还原状态、质子转移等很敏感-简而言之，人们需要知道许多事情来了解蛋白质功能和结构-功能关系。我们还建议使用其他技术，并根据需要开发新技术，以回答重要的科学问题。 在即将到来的赠款期间，我们将专注于血红素-铜氧化酶，其中有关为有氧生命提供动力的能量转导过程的关键问题仍未得到回答。我们提出了以下具体目标： 目标1：电子转移。氧还原和氧化还原连接的质子在血红素铜氧化酶中的传输：这些都是好氧生物能量学中的关键过程。我们将阐明活性部位、蛋白质结构和蛋白内水在氧化酶作用过程中的相互作用。具体项目包括(A)氧还原的红外(IR)流动-闪光研究，(B)同位素取代和突变研究，(C)蛋白质内电子转移研究，以及(D)蛋白质内水和质子传输研究。 目标2：技术开发，这是支持一些拟议项目所必需的。这包括(A)微流控和红外显微镜在时间分辨红外光谱中的应用，(B)红外光谱在单层蛋白质电化学中的应用，以及(C)核共振振动光谱(NRVS)在血红素-铜氧化酶中的应用。

DESCRIPTION (provided by applicant): The general objective of this project is to contribute to understanding of function and structure-function relationships in redox metalloproteins, primarily via observations of spectroscopy and dynamics. Our approach emphasizes vibrational (including time-resolved) spectroscopies, which are sensitive to conformations, redox states, proton transfers, etc. - in short, many of the things one needs to know to understand protein function and structure-function relationships. We also propose to use other techniques and develop new technologies as necessary to answer important scientific questions. During the upcoming grant period we will concentrate on the heme-copper oxidases, where key questions about the energy transduction processes that power aerobic life remain unanswered. We propose the following Specific Aims: Aim 1: Electron transfer. O2 reduction, and redox-linked proton transport in heme-copper oxidases: These are the key processes in aerobic bioenergetics. We will elucidate the interactive roles of the active sites, protein structure, and intraprotein water during the function of the oxidases. Specific projects include (a) infrared (IR) flow-flash studies of O2 reduction, (b) isotopic substitution and mutant studies, (c) studies of intraprotein electron transfer, and (d) studies of intraprotein water and proton transport. Aim 2: Technology development, which is required to support some of the projects proposed. This includes (a) Applications of microfluidics and IR microscopy to time-resolved IR, (b) IR spectroscopy applied to monolayer protein electrochemistry, and (c) nuclear resonance vibrational spectroscopy (NRVS) applied to heme-copper oxidases.