Liesener André, Karst Uwe
Chemical Analysis Group and MESA+ Institute for Nanotechnology, University of Twente, PO Box 217, 7500, AE Enschede, The Netherlands.
Anal Bioanal Chem. 2005 Aug;382(7):1451-64. doi: 10.1007/s00216-005-3305-2. Epub 2005 Jul 9.
This review highlights recent advances in the application of electrospray ionisation and matrix-assisted laser desorption/ionisation mass spectrometry (MS) to study enzymatic reactions. Several assay schemes for different fields of application are presented. The employment of MS as a means of detection in pre-steady-state kinetic studies by rapid-mixing direct analysis and rapid-mixing quench flow techniques is discussed. Several steady-state kinetic studies of a broad range of different enzymatic systems are presented as well as enzyme inhibition studies for various target enzymes. As a promising new development multiplex assays, which monitor the conversion of several substrates simultaneously in one experiment, are described. This assay type has been used for competition studies, enzymatic activity screenings and for diagnostic purposes in clinical chemistry. Generally, it can be concluded that mass spectrometry offers an intriguing alternative as detection methodology in enzymatic bioassays. Its applicability for the monitoring the conversion of naturally occurring substrates and its overall versatility make MS an especially promising tool for the study of enzyme-catalysed processes.
本综述重点介绍了电喷雾电离和基质辅助激光解吸/电离质谱（MS）在研究酶促反应中的最新应用进展。介绍了适用于不同应用领域的几种检测方案。讨论了在预稳态动力学研究中通过快速混合直接分析和快速混合淬灭流动技术将MS用作检测手段的情况。介绍了多种不同酶系统的几个稳态动力学研究以及针对各种靶酶的酶抑制研究。作为一个有前景的新发展方向，还描述了在一个实验中同时监测几种底物转化的多重检测。这种检测类型已用于竞争研究、酶活性筛选以及临床化学中的诊断目的。总的来说，可以得出结论，质谱作为酶促生物测定中的检测方法提供了一种有趣的替代方案。其在监测天然底物转化方面的适用性及其整体多功能性使MS成为研究酶催化过程的特别有前景的工具。
