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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.
Recently Published Articles
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Version of Record online: 27 APR 2017 | DOI: 10.1002/cbic.201700066
Modulation of sialic acid biosynthesis: Several drug-like small molecules have been identified as inhibitors of N-acetylmannosamine kinase by high-throughput screening. Validation and molecular modelling show high affinity in the low micromolar range for this important enzyme involved in the biosynthesis of sialic acids.
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Version of Record online: 27 APR 2017 | DOI: 10.1002/cbic.201700093
Transcriptome labelling: We describe a metabolic labelling approach for rapid generation of 13C bio-isotopologues of the C. elegans transcriptome to study its modifications by liquid-chromatography tandem high-resolution mass spectrometry (LC-HRMS). We identify the most abundant RNA modifications, study the response to environmental stress and demonstrate their dynamic behaviour.
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Jeremias E. G. A. Dold, Jessica Pfotzer, Dr. Anne-Katrin Späte and Prof. Dr. Valentin Wittmann
Version of Record online: 27 APR 2017 | DOI: 10.1002/cbic.201700002
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Version of Record online: 27 APR 2017 | DOI: 10.1002/cbic.201700154
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Version of Record online: 27 APR 2017 | DOI: 10.1002/cbic.201700033
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