title: A Clickable Aminooxy Probe for Monitoring Cellular ADP-Ribosylation.
abstract: ADP-ribosylation is essential for cell function, yet there is a dearth of methods for detecting this post-translational modification in cells. Here, we describe a clickable aminooxy alkyne (AO-alkyne) probe that can detect cellular ADP-ribosylation on acidic amino acids following Cu-catalyzed conjugation to an azide-containing reporter. Using AO-alkyne, we show that PARP10 and PARP11 are auto-ADP-ribosylated in cells. We also demonstrate that AO-alkyne can be used to monitor stimulus-induced ADP-ribosylation in cells. Functional studies using AO-alkyne support a previously unknown mechanism for ADP-ribosylation on acidic amino acids, wherein a glutamate or aspartate at the initial C1'-position of ADP-ribose transfers to the C2' position. This new mechanism for ADP-ribosylation has important implications for how glutamyl/aspartyl-ADP-ribose is recognized by proteins in cells.
authors.forename: Rory K
authors.initials: RK
authors.lastname: Morgan
authors.forename: Michael S
authors.initials: MS
authors.lastname: Cohen
mesh_terms: D000246:Adenosine Diphosphate Ribose / Q000032:analysis* / Q000378:metabolism*; D000480:Alkynes / Q000737:chemistry* / Q000378:metabolism; D001386:Azides / Q000737:chemistry* / Q000378:metabolism; D002384:Catalysis; D057930:Click Chemistry; D003300:Copper / Q000737:chemistry; D006801:Humans; D015335:Molecular Probes / Q000737:chemistry* / Q000378:metabolism; D011065:Poly(ADP-ribose) Polymerases / Q000032:analysis / Q000378:metabolism; D011499:Protein Processing, Post-Translational; D011506:Proteins / Q000378:metabolism*; D011518:Proto-Oncogene Proteins / Q000032:analysis / Q000378:metabolism
chemical_list: D000480:Alkynes; D001386:Azides; D015335:Molecular Probes; D011506:Proteins; D011518:Proto-Oncogene Proteins; D000246:Adenosine Diphosphate Ribose; D003300:Copper; C499627:PARP10 protein, human; D011065:Poly(ADP-ribose) Polymerases; C000605039:poly(ADP-ribose) polymerase-11, mouse