Journal article
Detection of Sulfoquinovosidase Activity in Cell Lysates Using Activity-Based Probes
Z Li, IB Pickles, M Sharma, B Melling, L Pallasdies, JDC Codée, SJ Williams, HS Overkleeft, GJ Davies
Angewandte Chemie International Edition | Published : 2024
Abstract
The sulfolipid sulfoquinovosyl diacylglycerol (SQDG), produced by plants, algae, and cyanobacteria, constitutes a major sulfur reserve in the biosphere. Microbial breakdown of SQDG is critical for the biological utilization of its sulfur. This commences through release of the parent sugar, sulfoquinovose (SQ), catalyzed by sulfoquinovosidases (SQases). These vanguard enzymes are encoded in gene clusters that code for diverse SQ catabolic pathways. To identify, visualize and isolate glycoside hydrolase CAZY-family 31 (GH31) SQases in complex biological environments, we introduce SQ cyclophellitol-aziridine activity-based probes (ABPs). These ABPs label the active site nucleophile of this enzy..
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Grants
Awarded by European Commission
Funding Acknowledgements
We thank the Royal Society (Ken Murray Research Professorship to G.J.D.), the Biotechnology and Biological Sciences Research Council (BBSRC) (grant BB/W003805/1 to G.J.D. and M.S.), the Netherlands Organization for Scientific Research (NWO TOP grant 2018-714.018.002 to H.S.O.), the European Research Council (ERC-2011-AdG-290836 "Chembiosphing" to H.S.O. and ERC-2020-SyG-951231 "Carbocentre" to G.J.D. and H.S.O.), and the Australian Research Council (DP210100233, DP210100235, DP240100126 to S.J.W.). Mass spectrometry was performed at the York Centre of Excellence in Mass Spectrometry, which benefited from capital investment through Science City York, supported by Yorkshire Forward with funds from the Northern Way Initiative, and from the EPSRC (EP/K039660/1; EP/M028127/1). We thank Hans van den Elst (University of Leiden) and Andrew Leech, Adam Dowle, Chris Taylor, Chloe Baldreki, Sally James, Lesley Gilbert and Samantha Donninger for technical support and advice (all University of York Bioscience Technology Facility). We thank Diamond Light Source for beamtime (proposals 24948, and 18598), and Johan Turkenburg, Sam Hart and the staff of beamline I03 for assistance with crystal testing and data collection.