Precursor ion scanning and sequencing of arginine-ADP-ribosylated peptide by mass spectrometry.
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.
Osago
Harumi
H
Department of Biochemistry, Shimane University, Faculty of Medicine, Izumo, Japan. biochem1@med.shimane-u.ac.jp
Yamada
Kazuo
K
Shibata
Tomoko
T
Yoshino
Ken-ichi
K
Hara
Nobumasa
N
Tsuchiya
Mikako
M
D000246:Adenosine Diphosphate Ribose / Q000031:analogs & derivatives* / Q000032:analysis; D053000:Analytic Sample Preparation Methods / Q000379:methods*; D009952:Ornithine / Q000032:analysis; D010455:Peptides / Q000737:chemistry*; D011064:Poly Adenosine Diphosphate Ribose / Q000737:chemistry; D011499:Protein Processing, Post-Translational; D011506:Proteins / Q000737:chemistry*; D020539:Sequence Analysis, Protein / Q000379:methods*; D053719:Tandem Mass Spectrometry / Q000379:methods*
C543491:ADP-ribosyl-carbodiimide; D010455:Peptides; D011506:Proteins; C024857:poly(ADP)-ribosylated proteins; D000246:Adenosine Diphosphate Ribose; D011064:Poly Adenosine Diphosphate Ribose; C043357:ADP-ribosylarginine; D009952:Ornithine