Metalloenzyme Mechanisms

超出所提供的空间。通过本研究应用的广泛、长期目标，我们试图了解生物大分子在大分子组装形成过程中识别其伙伴的机制。根据以下目标，建议对生物系统中识别事件的主要类别进行系统研究。(1)蛋白质-蛋白质识别。在这里，我们试图定义特定表面相互作用的作用；通过表面互补提供的静电、氢键和疏水自由能，决定了真核细胞色素P450依赖加氧酶中金属蛋白之间协调电子转移事件形成复合物的特异性和亲和性。(2)膜组分识别。生物膜具有磷脂、脂肪酸、胆固醇和其他关键细胞成分的复杂组成，它不仅仅是嵌入真核生物P450蛋白成分的简单“洗涤剂”。双分子层的组成和物理性质对于作为Aim(1)主题的识别事件非常重要。多种技术被用于理解膜结构对分子识别的贡献。(3)蛋白质小分子识别。在这个目的中，我们试图确定静电、氢键和底物与酶活性位点的紧密配合的基本力如何在细胞色素P450催化中产生观察到的高度区域和立体特异性控制。(4)蛋白质-核酸识别。同样的基本力量控制着识别过程。在这个目标中，我们建立在以前的成功定义的作用，溶剂水在调解蛋白质和核酸组分之间的氢键识别。我们通过研究有趣的“间接读出”大分子组装，并使用新颖的流体静力学和渗透压方法，结合晶体学和分子动力学模拟，扩大了系统研究的范围。我们提出的研究计划在这个竞争性的更新中协调一致地使用广泛的跨学科工具和技术，在分子生物学和先进的生物物理方法中，这些方法已被证明是理解金属酶机制的基本机制的理想选择。PERFORMANCESITE ( ======================================== 节结束 ===========================================

EXCEED THE SPACE PROVIDED. Through the broad, long term objectives of this research application, we seek to understand the mechanisms by which biological macromolecules recognize their partners in the formation of macromolecular assemblies. Systematic investigations are proposed that focus on the major classes of recognition events in biological systems according to the following Aims. (1) Protein - Protein Recognition. Here we seek to define the role of specific surface interactions; electrostatic, hydrogen bonding, and hydrophobic free energies provided through surface complimentary, which define the specificity and affinity in the formation of complexes between the metalloproteins coordinating electron transfer events in the eukaryotic cytochrome P450 dependent oxygenases. (2) Membrane component recognition. Biological membranes, with their complex makeup of phospholipids, fatty acids, cholesterol and other critical cellular components, is more than a simple "detergent" for embedded eukaryotic P450 protein components. The composition and physical properties of the bilayer are important for the very recognition events that are the subject of Aim (1). Multiple techniques are brought to bear toward the goal of understanding the contributions of membrane architectures to molecular recognition. (3) Protein - Small Molecule Recognition. In this Aim we seek to ascertain how the same fundamental forces of electrostatics, hydrogen bonding and the hand - glove fit of a substrate into the active site of an enzyme can give rise to the observed high degree control of regio- and stereo- specificity in cytochrome P450 catalysis. (4) Protein - Nucleic Acid Recognition. Again the same fundamental forces control recognition processes. In this Aim we build on previous success in defining roles of solvent water in mediating hydrogen bond recognition between protein and nucleic acid components. We expand the repertoire of systems investigated by examining the interesting class of "indirect readout" macromolecular assemblies and use novel hydrostatic and osmotic pressure methods, in concert with crystallography and molecular dynamics simulations. Our proposed research program in this competitive renewal make concerted use of broad interdisciplinary tools and techniques in molecular biology and advanced biophysical methods which have proven to be ideal for understanding the fundamental mechanisms of metalloenzyme mechanisms. PERFORMANCESITE( ========================================Section End===========================================