biostudies-literatureSchum DNational Institutes of HealthNIGMS NIH HHSNIH HHSe0054025https://www.ebi.ac.uk/biostudies/studies/S-EPMC12172467biostudies-literatureUnknown10(6)Pyridoxal 5'-phosphate (PLP) is an essential cofactor for enzymes that catalyze diverse reactions in central metabolism. 2-Aminoacrylate (2AA) is a reactive enamine and an obligate catalytic intermediate in some PLP-mediated reactions. In the absence of the enamine/imine deaminase RidA, <i>Salmonella enterica</i> accumulates 2AA, which causes cellular stress. 2AA can attack PLP in the active site of some enzymes and covalently inactivate them by forming a 2AA-PLP adduct, which has already been characterized for some target enzymes <i>in vivo</i> and <i>in vitro</i>. The mechanism of 2AA attack suggests that a majority of cellular PLP-DEs would be targets of 2AA damage. Herein, a chemical proteomics workflow that uses PL (pyridoxal) probes to enrich PLP-DEs with a click chemistry-based protmSystemsChemical proteomics enhances the understanding of 2AA stress in &lt;i&gt;Salmonella enterica&lt;/i&gt;.PMC12172467RO1GM095837R35 GM153189R35GM153189R01 GM095837Schum DSieber SAFiedler MKShen WDowns DMfalseChemical proteomics enhances the understanding of 2AA stress in &lt;i&gt;Salmonella enterica&lt;/i&gt;.Pyridoxal 5'-phosphate (PLP) is an essential cofactor for enzymes that catalyze diverse reactions in central metabolism. 2-Aminoacrylate (2AA) is a reactive enamine and an obligate catalytic intermediate in some PLP-mediated reactions. In the absence of the enamine/imine deaminase RidA, <i>Salmonella enterica</i> accumulates 2AA, which causes cellular stress. 2AA can attack PLP in the active site of some enzymes and covalently inactivate them by forming a 2AA-PLP adduct, which has already been characterized for some target enzymes <i>in vivo</i> and <i>in vitro</i>. The mechanism of 2AA attack suggests that a majority of cellular PLP-DEs would be targets of 2AA damage. Herein, a chemical proteomics workflow that uses PL (pyridoxal) probes to enrich PLP-DEs with a click chemistry-based prot2025-01-01T00:00:00Z2025 Jun2026-06-03T07:08:42.575Z2026-04-25T03:22:19.817ZS-EPMC121724674043940910.1128/msystems.00540-25