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dc.contributor.authorHübner, Ines
dc.contributor.authorDienemann, Jan-Niklas
dc.contributor.authorFriederich, Julia
dc.contributor.authorSchneider, Sabine
dc.contributor.authorSieber, Stephan A
dc.date.accessioned2021-01-13T09:18:11Z
dc.date.available2021-01-13T09:18:11Z
dc.date.issued2020-12-03
dc.identifier.citationACS Chem Biol. 2020 Dec 18;15(12):3227-3234. doi: 10.1021/acschembio.0c00787. Epub 2020 Dec 3.en_US
dc.identifier.pmid33269909
dc.identifier.doi10.1021/acschembio.0c00787
dc.identifier.urihttp://hdl.handle.net/10033/622675
dc.description.abstractPyridoxal kinases (PLK) are crucial enzymes for the biosynthesis of pyridoxal phosphate, an important cofactor in a plethora of enzymatic reactions. The evolution of these enzymes resulted in different catalytic designs. In addition to the active site, the importance of a cysteine, embedded within a distant flexible lid region, was recently demonstrated. This cysteine forms a hemithioacetal with the pyridoxal aldehyde and is essential for catalysis. Despite the prevalence of these enzymes in various organisms, no tools were yet available to study the relevance of this lid residue. Here, we introduce pyridoxal probes, each equipped with an electrophilic trapping group in place of the aldehyde to target PLK reactive lid cysteines as a mimic of hemithioacetal formation. The addition of alkyne handles placed at two different positions within the pyridoxal structure facilitates enrichment of PLKs from living cells. Interestingly, depending on the position, the probes displayed a preference for either Gram-positive or Gram-negative PLK enrichment. By applying the cofactor traps, we were able to validate not only previously investigated Staphylococcus aureus and Enterococcus faecalis PLKs but also Escherichia coli and Pseudomonas aeruginosa PLKs, unravelling a crucial role of the lid cysteine for catalysis. Overall, our tailored probes facilitated a reliable readout of lid cysteine containing PLKs, qualifying them as chemical tools for mining further diverse proteomes for this important enzyme class.en_US
dc.language.isoenen_US
dc.publisherAmerican Society for Chemistry (ACS)en_US
dc.rightsAttribution-NonCommercial-ShareAlike 4.0 International*
dc.rights.urihttp://creativecommons.org/licenses/by-nc-sa/4.0/*
dc.titleTailored Cofactor Traps for the Detection of Hemithioacetal-Forming Pyridoxal Kinases.en_US
dc.typeArticleen_US
dc.identifier.eissn1554-8937
dc.contributor.departmentHZI,Helmholtz-Zentrum für Infektionsforschung GmbH, Inhoffenstr. 7,38124 Braunschweig, Germany.en_US
dc.identifier.journalACS chemical biologyen_US
dc.source.volume15
dc.source.issue12
dc.source.beginpage3227
dc.source.endpage3234
dc.source.journaltitleACS chemical biology
dc.source.countryUnited States


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