Identification of GlcNAcylation sites of peptides and alpha-crystallin using Q-TOF mass spectrometry.
The addition of a single N-acetylglucosamine residue O-linked to serine and threonine residues of nuclear and cytoplasmic proteins is a widespread modification throughout all eukaryotes. The conventional method for detecting and locating sites of modification is a multi-step radioactivity-based protocol. In this paper we show that using quadrupole time-of-flight (Q-TOF) mass spectrometry, modification sites can be identified at a significantly higher sensitivity than previous approaches. This is the first demonstration that sites of O-GlcNAcylation can be identified directly using mass spectrometry.
Chalkley
R J
RJ
Ludwig Institute for Cancer Research, University College Branch of Cell and Molecular Biology, Royal Free and University College Medical School, London, UK.
Burlingame
A L
AL
D000215:Acylation; D000595:Amino Acid Sequence; D002851:Chromatography, High Pressure Liquid; D003459:Crystallins / Q000737:chemistry*; D004591:Electrophoresis, Polyacrylamide Gel; D005690:Galactose / Q000737:chemistry; D006868:Hydrolysis; D013058:Mass Spectrometry; D008969:Molecular Sequence Data; D010455:Peptides / Q000737:chemistry*; D021241:Spectrometry, Mass, Electrospray Ionization
D003459:Crystallins; D010455:Peptides; D005690:Galactose