A Novel Instrument for Studying Transient Enzyme Intermediates

了解酶催化如何在酶活性位点发生，可以为蛋白质结构-功能研究和基于结构的药物设计提供关键见解。催化作用的研究是理解酶功能的核心。对酶活性位点内发生的化学、化学物种和动力学途径的理解，对于详细描述酶的工作原理，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。我们正在开发的这一仪器和方法的主要重点将是推进生物化学反应中的瞬态中间体（包括共价和非共价）的研究。此外，该仪器应该在监测化学催化和反应，帮助蛋白质折叠结构和动力学研究以及进一步验证和表征具有新催化功能的工程蛋白的创造方面发挥作用。该仪器将开辟研究化学不稳定的中间体和新的酶和蛋白质系统的可能性，否则无法研究。该仪器将为瞬态酶中间体的研究方法提供重大进展，并将继续将高分辨率质谱技术应用于生物学。随着方法变得更加强大，随着具有新机制的新酶被发现，随着将瞬态动力学信息纳入药物设计的增加以及快速动力学研究（如蛋白质折叠）的新领域的出现，这一领域的重要性将继续增长。我们现在正在见证质谱法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功率分析检测相结合，有可能将酶学推向一个新的水平。

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.