A Novel Instrument for Studying Transient Enzyme Intermediates

研究瞬时酶中间体的新型仪器

• 批准号: 9876677
• 负责人: Karen Anderson
• 金额: $ 35万
• 依托单位: Yale University
• 依托单位国家: 美国
• 项目类别: Continuing Grant
• 财政年份: 1999
• 资助国家: 美国
• 起止时间: 1999-04-01 至 2003-03-31
• 项目状态: 已结题
• 来源: https://www.nsf.gov/awardsearch/showAward?AWD_ID=9876677&HistoricalAwards=false
• 关键词: Novel; Instrument; Studying; Transient; Enzyme

Key insights into protein structure-function studies and structure-based drug design can be provided by an understanding of how enzyme catalysis is occurring at the enzyme active site. The study of catalysis is at the very core of understanding enzyme function. An understanding of the chemistry, chemical species and the kinetic pathway that takes place within the active site of enzyme holds great promise for describing in detail how enzymes work and more importantly how small molecules might be designed to be more potent drugs i.e. transition state analogs. The major focus of this instrument and methodology we are developing will be in advancing studies of transient intermediates in biochemical reactions (both covalent and non-covalent). In addition, this instrument should find utility in monitoring chemical catalysis and reactions, aiding in protein folding structural and kinetic studies and further validating and characterizing the creation of engineered proteins with novel catalytic functions. This instrument will open up the possibility of studying chemically labile intermediates and new enzyme and protein Systems that otherwise could not be studied. This instrument will provide a major advance in methods to study transient enzyme intermediates and will continue the evolution of applying high-resolution mass spectrometry techniques into biology. This area continues to grow in importance as the methods become more powerful, as new enzymes with novel mechanisms become identified, as incorporation of transient kinetic information into drug design increases and as new areas of rapid kinetic investigations such as protein folding emerge. We are, just now, witnessing the power and impact that mass spectrometry will have in the biological sciences. This instrument, which interfaces classical rapid kinetics studies of enzymes with high powered analytical detection, has the possibility of propelling enzymology to a new level.

通过了解酶催化如何在酶活性位点发生，可以提供对蛋白质结构功能研究和基于结构的药物设计的关键见解。催化研究是理解酶功能的核心。了解酶活性位点内发生的化学、化学种类和动力学途径，对于详细描述酶的工作原理以及更重要的是如何将小分子设计成更有效的药物（即过渡态类似物）具有很大的希望。我们正在开发的该仪器和方法的主要重点将是推进生化反应（共价和非共价）中瞬时中间体的研究。此外，该仪器应可用于监测化学催化和反应，帮助蛋白质折叠结构和动力学研究，并进一步验证和表征具有新颖催化功能的工程蛋白质的产生。该仪器将为研究化学不稳定中间体以及新的酶和蛋白质系统开辟可能性，否则无法研究这些系统。该仪器将为研究瞬时酶中间体的方法提供重大进步，并将继续将高分辨率质谱技术应用于生物学的发展。随着方法变得更加强大，随着具有新颖机制的新酶的被发现，随着瞬态动力学信息融入药物设计的增加以及随着快速动力学研究的新领域（例如蛋白质折叠）的出现，该领域的重要性继续增长。我们现在正在见证质谱分析在生物科学中的力量和影响。该仪器将酶的经典快速动力学研究与高性能分析检测相结合，有可能将酶学推向一个新的水平。