Precursor ion scanning and sequencing of arginine-ADP-ribosylated peptide by mass spectrometry

• Published in: Analytical biochemistry 2009-10, Vol.393 (2), p.248-254
• Main Authors: Osago, Harumi, Yamada, Kazuo, Shibata, Tomoko, Yoshino, Ken-ichi, Hara, Nobumasa, Tsuchiya, Mikako
• Format: Article
• Language: English
• Subjects: Adenosine Diphosphate Ribose - analogs & derivatives; Adenosine Diphosphate Ribose - analysis; Analytic Sample Preparation Methods - methods; Arginine-specific ADP-ribosylation; Mass spectrometry; N-(ADP-ribosyl)-carbodiimide; Ornithine; Ornithine - analysis; Peptides - chemistry; Poly Adenosine Diphosphate Ribose - chemistry; Posttranslational modification; Precursor ion scanning; Protein Processing, Post-Translational; Proteins - chemistry; Sequence Analysis, Protein - methods; Tandem Mass Spectrometry - methods
• ISSN: 0003-2697; 1096-0309
• DOI: 10.1016/j.ab.2009.06.028

Arginine (Arg)-specific ADP-ribosylation is one of the posttranslational modifications of proteins and is thought to play an important role in reversibly regulating functions of the target proteins in eukaryotes. However, the physiological target protein has not been established. We examined the fragmentation pattern of both ADP-ribosyl-Arg (ADP-R-Arg) and Arg-ADP-ribosylated peptides by quadrupole tandem mass spectrometry and found a specific cleavage of ADP-R-Arg into N-(ADP-ribosyl)-carbodiimide (ADP-R-carbodiimide) and ornithine. Based on this specific fragmentation pattern, we successfully identified the modification site and sequence of Arg-ADP-ribosylated peptide using a two-step collision and showed that ADP-R-carbodiimide is an excellent marker ion for precursor ion scanning of Arg-ADP-ribosylated peptide. We propose that a combination of the precursor ion scanning with ADP-R-carbodiimide as a marker ion and two-step collision is useful in searching for physiological target proteins of Arg-ADP-ribosylation.