title: Chemoproteomic profiling and discovery of protein electrophiles in human cells.
abstract: Activity-based protein profiling (ABPP) serves as a chemical proteomic platform to discover and characterize functional amino acids in proteins on the basis of their enhanced reactivity towards small-molecule probes. This approach, to date, has mainly targeted nucleophilic functional groups, such as the side chains of serine and cysteine, using electrophilic probes. Here we show that 'reverse-polarity' (RP)-ABPP using clickable, nucleophilic hydrazine probes can capture and identify protein-bound electrophiles in cells. Using this approach, we demonstrate that the pyruvoyl cofactor of S-adenosyl-L-methionine decarboxylase (AMD1) is dynamically controlled by intracellular methionine concentrations. We also identify a heretofore unknown modification-an N-terminally bound glyoxylyl group-in the poorly characterized protein secernin-3. RP-ABPP thus provides a versatile method to monitor the metabolic regulation of electrophilic cofactors and discover novel types of electrophilic modifications on proteins in human cells.
authors.affiliation: Chemical Physiology, The Scripps Research Institute, La Jolla, California 92037, USA.
authors.forename: Megan L
authors.identifier: 0000-0003-1178-4616
authors.initials: ML
authors.lastname: Matthews
authors.affiliation: Chemical Physiology, The Scripps Research Institute, La Jolla, California 92037, USA.|Bioinformatics Solutions Inc., Waterloo, Ontario N2L 6J2, Canada.
authors.forename: Lin
authors.initials: L
authors.lastname: He
authors.affiliation: Chemical Physiology, The Scripps Research Institute, La Jolla, California 92037, USA.
authors.forename: Benjamin D
authors.initials: BD
authors.lastname: Horning
authors.affiliation: Chemistry and Cell and Molecular Biology, The Scripps Research Institute, La Jolla, California 92037, USA.
authors.forename: Erika J
authors.initials: EJ
authors.lastname: Olson
authors.affiliation: Chemical Physiology, The Scripps Research Institute, La Jolla, California 92037, USA.|École polytechnique fédérale de Lausanne, CH-1015 Lausanne, Switzerland.
authors.forename: Bruno E
authors.initials: BE
authors.lastname: Correia
authors.affiliation: Chemical Physiology, The Scripps Research Institute, La Jolla, California 92037, USA.
authors.forename: John R
authors.initials: JR
authors.lastname: Yates
authors.affiliation: Chemistry and Cell and Molecular Biology, The Scripps Research Institute, La Jolla, California 92037, USA.
authors.forename: Philip E
authors.initials: PE
authors.lastname: Dawson
authors.affiliation: Chemical Physiology, The Scripps Research Institute, La Jolla, California 92037, USA.
authors.forename: Benjamin F
authors.initials: BF
authors.lastname: Cravatt
mesh_terms: D012437:Adenosylmethionine Decarboxylase / Q000737:chemistry* / Q000378:metabolism; D000596:Amino Acids / Q000737:chemistry / Q000378:metabolism; D002470:Cell Survival; D005456:Fluorescent Dyes / Q000737:chemistry; D057809:HEK293 Cells; D006801:Humans; D006834:Hydrazines / Q000737:chemistry; D008715:Methionine / Q000737:chemistry / Q000378:metabolism; D015394:Molecular Structure; D009419:Nerve Tissue Proteins / Q000737:chemistry* / Q000378:metabolism; D040901:Proteomics
chemical_list: D000596:Amino Acids; D005456:Fluorescent Dyes; D006834:Hydrazines; D009419:Nerve Tissue Proteins; C029424:hydrazine; D008715:Methionine; C000608756:AMD1 protein, human; D012437:Adenosylmethionine Decarboxylase