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Monitoring enzymatic conversions by mass spectrometry: a critical review

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Abstract

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.

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  1. Chemical Analysis Group and MESA+ Institute for Nanotechnology, University of Twente, PO Box 217, 7500, AE Enschede, The Netherlands

    André Liesener & Uwe Karst

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  1. André Liesener
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  2. Uwe Karst
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Correspondence to Uwe Karst.

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Liesener, A., Karst, U. Monitoring enzymatic conversions by mass spectrometry: a critical review. Anal Bioanal Chem 382, 1451–1464 (2005). https://doi.org/10.1007/s00216-005-3305-2

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