Atomic level mutagenesis to study hydrogen bonds

研究氢键的原子水平诱变

• 批准号: 6998026
• 负责人: JASON P SCHWANS
• 金额: $ 4.4万
• 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/6998026
• 关键词: active sites; aspartate; bioenergetics; chemical synthesis; enzyme activity; gene mutation; hydrogen bond; isomerase; oxysteroid; postdoctoral investigator; protein structure function; site directed mutagenesis

DESCRIPTION (provided by applicant): 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 (Asp 103) 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.

描述（由申请人提供）：