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March 14, 2017
Very Important Paper: Identifying Unknown Enzyme-Substrate Pairs from the Cellular Milieu with Native Mass Spectrometry
Kalli C. Catcott, Jing Yan, Wanlu Qu, Vicki H. Wysocki*, Zhaohui Sunny Zhou*
The enzyme-substrate complex is inherently transient, rendering its detection difficult. Therefore, V. Wysocki (The Ohio State University, U.S.A.), Z.S. Zhou (Northeastern University, USA) and co-workers developed a framework, IsoLAIT (Isotope-Labeled, Activity-based Identification and Tracking), for bisubstrate systems. Using this method a common substrate, such as S-adenosylmethionine for methyltransferases, is replaced by an analogue (e.g., S-adenosylvinthionine) that, as a probe, creates a tightly bound [enzyme•substrate-probe] complex upon catalysis by thiopurine-S-methyltransferase. Then, this persistent complex is identified by native mass spectrometry from the cellular milieu without separation. Furthermore, the probe’s isotope pattern flags even unknown substrates and enzymes. IsoLAIT can be broadly applicable for other enzyme systems, particularly those catalyzing group transfer and with multiple substrates.
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