Identification of ADP-ribosylation sites of CD38 mutants by precursor ion scanning mass spectrometry Academic Article uri icon


MeSH Major

  • Antigens, CD38
  • Membrane Glycoproteins
  • Mutation, Missense
  • Poly Adenosine Diphosphate Ribose
  • Protein Processing, Post-Translational
  • Proteins


  • Protein ADP-ribosylation, including mono- and poly-ADP-ribosylation, is increasingly recognized to play important roles in various biological pathways. Molecular understanding of the functions of ADP-ribosylation requires the identification of the sites of modification. Although tandem mass spectrometry (MS/MS) is widely recognized as an effective means for determining protein modifications, identification of ADP-ribosylation sites has been challenging due to the labile and hydrophilic nature of the modification. Here we applied precursor ion scanning-triggered MS/MS analysis on a hybrid quadrupole linear ion trap mass spectrometer for selectively detecting ADP-ribosylated peptides and determining the auto-ADP-ribosylation sites of CD38 (cluster of differentiation 38) E226D and E226Q mutants. CD38 is an enzyme that catalyzes the hydrolysis of nicotinamide adenine dinucleotide (NAD) to ADP-ribose. Here we show that NAD can covalently label CD38 E226D and E226Q mutants but not wild-type CD38. In this study, we have successfully identified the D226/Q226 and K129 residues of the two CD38 mutants being the ADP-ribosylation sites using precursor ion scanning hybrid quadrupole linear ion trap mass spectrometry. The results offer insights about the CD38 enzymatic reaction mechanism. The precursor ion scanning method should be useful for identifying the modification sites of other ADP-ribosyltransferases such as poly(ADP-ribose) polymerases.

publication date

  • February 15, 2013



  • Academic Article



  • eng

PubMed Central ID

  • PMC3601590

Digital Object Identifier (DOI)

  • 10.1016/j.ab.2012.10.029

PubMed ID

  • 23123429

Additional Document Info

start page

  • 218

end page

  • 26


  • 433


  • 2