title: A chemical reporter for protein AMPylation.
abstract: Protein AMPylation is an emerging post-translational modification, which plays key roles in bacterial pathogenesis and cell biology. Enzymes with AMPylation activity, referred to as AMPylators, have been identified in several bacterial pathogens and eukaryotes. To facilitate the study of this unique modification, we developed an alkynyl chemical reporter for detection and identification of protein AMPylation substrates. Covalent functionalization of AMPylation substrates with the alkynyl reporter in lieu of adenylyl 5'-monophosphate (AMP) allows their subsequent bioorthogonal ligation with azide-fluorescent dyes or affinity enrichment tags. We show that this chemical reporter is transferred by a range of AMPylators onto their cognate protein substrates and allows rapid detection and identification of AMPylated substrates.
authors.affiliation: The Laboratory of Chemical Biology and Microbial Pathogenesis, The Rockefeller University, New York, New York 10065, USA.
authors.forename: Markus
authors.initials: M
authors.lastname: Grammel
authors.forename: Phi
authors.initials: P
authors.lastname: Luong
authors.forename: Kim
authors.initials: K
authors.lastname: Orth
authors.forename: Howard C
authors.initials: HC
authors.lastname: Hang
mesh_terms: D000249:Adenosine Monophosphate / Q000737:chemistry / Q000378:metabolism*; D000480:Alkynes / Q000737:chemistry / Q000378:metabolism*; D011506:Proteins / Q000737:chemistry / Q000378:metabolism*; D040901:Proteomics
chemical_list: D000480:Alkynes; D011506:Proteins; D000249:Adenosine Monophosphate