Atomic level mutagenesis to study hydrogen bonds

研究氢键的原子水平诱变

• 批准号: 7134305
• 负责人: JASON P SCHWANS
• 金额: $ 4.88万
• 依托单位: STANFORD UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2006
• 资助国家: 美国
• 起止时间: 2006-01-01 至 2007-12-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7134305
• 关键词: Accounting; Acids; Active Sites; Address; Affinity; Amino Acids; Aspartic Acid; Behavior; Binding; Biological Process; C-terminal; Catalysis; Chemicals; Classification; Complex; Crystallography; Cysteine; Depth; Discrimination; Electrostatics; Environment; Enzymes; Foundations; Hydrogen Bonding; Individual; Isomerase; Ketosteroids; Kinetics; Laboratories; Length; Ligation; Literature; Measures; Modeling; Mutagenesis; N-terminal; Nature; Organic Chemistry; Peptide Synthesis; Peptides; Personal Satisfaction; Phase; Phenols; Positioning Attribute; Procedures; Property; Proteins; Rate; Reaction; Relative (related person); Resolution; Role; Series; Site; Site-Directed Mutagenesis; Solid; Solutions; Specificity; System; Testing; Thermodynamics; Twin Multiple Birth; Variant; analog; aqueous; base; chemical property; cofactor; cysteine sulfinic acid; enzyme activity; enzyme structure; functional group; reaction rate; research study

Understanding how enzymes achieve their enormous catalytic power and exquisite specificity is central to the study of biological function. Over the past decades many basic features of enzyme function and behavior have been illuminated. Nevertheless, the extraordinary rate enhancements and specificites of enzymes cannot be accounted for quantitatively. Unnatural amino acids are required to probe at an unprecedented depth the energetic consequences of what is arguably the most profound difference between enzymatic and uncatalyzed solution reactions¿carrying out reactions in the idiosyncratic and highly specialized enzyme active site instead of aqueous solution. A series of ketosteroid isomerase variants will be generated in which the properties of an active site aspartic acid residue (Asp103) that donates a hydrogen bond are varied systematically and rationally. These studies will deepen our understanding of hydrogen bonding energetics within an enzyme active site and advance our understanding of the properties of the enzymatic environment that dictate the nature and energetics of such hydrogen bonds. These experiments will serve as a foundation for the use of systematic and incisive chemical perturbation in the study of enzymes.

了解酶如何实现其巨大的催化能力和精致的特异性是核心， 研究生物学功能。在过去的几十年里，酶的功能和行为的许多基本特征 被照亮了。然而，酶的非凡速率增强和特异性 无法定量计算。非天然氨基酸需要探测在一个前所未有的 深度什么可以说是最深刻的差异之间的活力后果酶和 非催化溶液反应，在特异性和高度专业化的酶中进行反应 活性位点而不是水溶液。将产生一系列酮类固醇异构酶变体，其中 提供氢键的活性位点天冬氨酸残基（Asp103）的性质是不同的 系统而合理地。这些研究将加深我们对氢键能量学的理解 在酶的活性位点和推进我们的酶环境的性质的理解 决定了这种氢键的性质和能量。这些实验将作为 在酶的研究中使用系统而深刻的化学扰动。