Journal of the American Society for Mass Spectrometry

, Volume 20, Issue 3, pp 477–483

Tandem mass spectrometry investigation of ADP-ribosylated kemptide

Authors

  • Shawna M. Hengel
    • Department of Medicinal ChemistryUniversity of Washington
  • Scott A. Shaffer
    • Department of Medicinal ChemistryUniversity of Washington
  • Brook L. Nunn
    • Department of Medicinal ChemistryUniversity of Washington
    • Department of Medicinal ChemistryUniversity of Washington
Article

DOI: 10.1016/j.jasms.2008.10.025

Cite this article as:
Hengel, S.M., Shaffer, S.A., Nunn, B.L. et al. J Am Soc Mass Spectrom (2009) 20: 477. doi:10.1016/j.jasms.2008.10.025

Abstract

Bacterial adenosine diphosphate-ribosyltransferases (ADPRTs) are toxins that play a significant role in pathogenicity by inactivating host proteins through covalent addition of ADP-ribose. In this study we used ADP-ribosylated Kemptide (LRRASLG) as a standard to examine the effectiveness of three common tandem mass spectrometry fragmentation methods for assignment of amino acid sequence and site of modification. Fragmentation mechanisms investigated include low-energy collision-induced dissociation (CID), infrared multiphoton dissociation (IRMPD), and electron-capture dissociation (ECD); all were performed on a hybrid linear ion trap Fourier transform ion cyclotron resonance mass spectrometer. We show that ECD, but neither CID nor IRMPD, of ADP-ribosylated Kemptide produces tandem mass spectra that are interpretable with regard to amino acid sequence assignment and site of modification. Examination of CID and IRMPD tandem mass spectra of ADP-ribosylated Kemptide revealed that fragmentation was primarily focused to the ADP-ribose region, generating several potential diagnostic ions for use in discovery of ADP-ribosylated proteins. Because of the lower relative sensitivity of ECD during data-dependent acquisition to CID, we suggest a 2-fold strategy where CID and IRMPD are first used to detect ADP-ribosylated peptides, followed by sequence assignment and location of modification by ECD analysis.

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© American Society for Mass Spectrometry 2009