MOLECULAR & CONTROL MECHANISM OF ENERGY COUPLING SITE I

我们建议研究电子和质子转移的机制。 线粒体NADH-醌氧化还原酶途径(通常被称为 如复杂的i)。这种络合物在氧化的过程中起着中心作用。 NADH，细胞代谢的还原产物，通过呼吸 链条。复合体I是最复杂和最不被理解的能量 呼吸链质子动力酶的转导。建议的 研究在机制的研究方面有了新的方向， 该复合体的动力学和调节特性在两个孤立的 和膜结合态。具体研究将包括：[i] 催化活性络合物I最小结构的测定 NADH氧化与质子的矢量转移相耦合 膜.[II]内部电子转移的测定 NADH-苯二酮氧化还原酶固有氧化还原成分的序列分析 利用人造电子受体(过渡金属络合物) 具有适当的氧化还原电位和适当的其他参数 (亲水性、电荷、大小)；[III]关于该化合物的物理化学研究 与配合物I中的特定结合位点相关的乌双喹酮类化合物 最近在俄罗斯公司S实验室发现的；[IV] 慢活/慢停的分子事件研究 复合体I的转变，最近被俄罗斯联合计划署重新发现， 这将扩展到对生理相关因素的搜索 它们参与了对这个综合体的滞回行为的控制。 私家侦探在EPR研究和热力学方面有丰富的经验 线粒体和线粒体固有顺磁性氧化还原中心的分析 微生物系统。具体地说，私家侦探已经做出了重大贡献 在表征铁-硫团簇、黄素和 泛醌自由基。俄罗斯公安局是一个老牌的 生物化学家，最近开发出了独特的亚线粒体颗粒 具有紧密耦合的I位能量转导的制剂。这个 因此，这些调查人员之间的合作计划是可望的 大大提高对能量耦合机制的认识 以及它的规定。

We propose to study the mechanism of electron and proton transfer pathways in mitochondrial NADH-quinone oxidoreductase (often referred to as Complex I). this complex plays a central role in the oxidation of NADH, the reducing product of cellular metabolism, by the respiratory chain. Complex I is the most complicated and least understood energy transducing proton-motive enzyme of the respiratory chain. the proposed research takes new directions in the investigation of the mechanism, kinetics, and regulatory properties of this complex in both the isolated and membrane-bound states. The specific studies will include: [I] determination of the minimal structure of Complex I capable of catalyzing NADH oxidation which is coupled with vectorial transfer of protons across the membrane; [II] determination of the internal electron transfer sequence within intrinsic redox components of NADH-quinone oxidoreductase by utilizing artificial electron acceptors (transition metal complexes) with proper redox potentials and appropriate other parameters (hydrophilicity, charge, size); [III] physico-chemical studies on the ubisemiquinones associated with the specific binding sites in Complex I which was recently discovered in Russian Co-P.I.'s laboratory; [IV] studies of molecular events involved in the slow active/inactive transition of Complex I, rediscovered recently by the Russian Co-P.I., which will be extended to searches of physiologically relevant factors which are involved in the control of hysteretic behavior of this complex. The P.I. has extensive experience with EPR studies and with thermodynamic analysis of the intrinsic paramagnetic redox centers of mitochondrial and microbial systems. Specifically, the P.I. has made significant contribution in the characterization of iron-sulfur clusters, flavin and ubiquinone free radicals. The Russian Co-P.I. is an established biochemist and has recently developed unique submitochondrial particle preparations having tightly coupled Site I energy transduction. The collaborative program between these investigators, therefore, is expected to greatly enhance the understanding of the mechanism of energy coupling and its regulation.