Presidential Young Investigator Award

F1 atp酶是一个大的膜相关蛋白，它连接两个腺嘌呤核苷酸，三磷酸腺苷和二磷酸腺苷之间的平衡反应，以及质子在膜上的易位。因此，通过膜上合适的质子电化学电位差，它可以驱动三磷酸腺苷的净合成。该体系将用磷固体核磁共振进行研究。将尝试检测反应中的中间体并测量反应速率。还将尝试确定反应所需的结合金属离子（锰将取代镁）与核苷酸磷酸基团之间的距离。这项工作的总体目标是确定酶的原位活性位点的结构和动力学；以前的研究依赖于提取技术，这可能不能反映酶本身的反应物的真实浓度。本研究的长期目标是扩展核磁共振实验，并测量蛋白质上特定氨基酸残基与结合的腺嘌呤核苷酸的接近程度。

The F1 ATPase is large, membrane-associated protein which links an equilibrium reaction between the two adenine nucleotides, adenosine triphosphate and adenosine diphosphate, and the translocation of protons across the membrane. Thus, with a suitable proton electrochemical potential difference across the membrane it can drive the net synthesis of adenosine triphosphate. This system will be studied by phosphorous solid state nuclear magnetic resonance. Attempts will be made to detect intermediates in the reaction and to measure rates of reaction. Attempts will also be made to determine distances between a bound metal ion required for reactions (manganese will be substituted for magnesium) and the phosphate groups of the nucleotide. %%% The overall goal of this work is to define the structure and kinetics of the active site of the enzyme in situ; previous studies have relied on extraction techniques which may not reflect the true concentrations of reactants on the enzyme itself. Longer term aims of this study are to extend the NMR experiments and measure the proximity of specific amino acid residues on the protein to the bound adenine nucleotides.