A Review of Tandem Mass Spectrometry Characterization of Adenosine Diphosphate-Ribosylated Peptides

Abstract

The use of tandem mass spectrometry to identify and characterize sites of protein adenosine diphosphate (ADP) ribosylation will be reviewed. Specifically, we will focus on data acquisition schemes and fragmentation techniques that provide peptide sequence and modification site information. Also discussed are uses of synthetic standards to aid characterization, and an enzymatic method that converts ADP-ribosylated peptides into ribosyl mono phosphorylated peptides making identification amenable to traditional phosphopeptide characterization methods. Finally the potential uses of these techniques to characterize poly ADP-ribosylation sites, and inherent challenges, are addressed.

Figure 1

A) Transfer of ADP-ribose from β-NAD+ to arginine is catalyzed by ADP-ribosyl transferases and B) Poly ADP-ribose polymerase (PARP-1) catalyzes ADP-ribosylation and subsequent ADP-ribose polymers (N′).

Figure 2

Representative tandem mass spectra of ADP-ribosylated peptides using A) CID or B) ECD fragmentation [16].

Figure 3

A) Proposed ADP-ribosylated peptide fragment ion nomenclature as suggested by Hengel et al., B) cross ring fragmentation annotation, and c) poly ADP-ribosylation annotation.

Figure 4

Phosphodiesterase (PDE) treatment of mono ADP-ribosylated peptide yields a ribose mono-phosphate peptide adduct.

Figure 5

A) CID tandem mass spectrum of PDE treated Kemptide, B) zoomed in m/z range of CID spectrum, and C) MSA CID spectrum of PDE treated ADP-ribosylated Kemptide.