Identification of novel sites of O-N-acetylglucosamine modification of serum response factor using quadrupole time-of-flight mass spectrometry.
The addition of a single N-acetylglucosamine moiety O-linked to serine and threonine residues of nuclear and cytoplasmic proteins is a widespread post-translational modification. The conventional method for detecting and locating sites of modification is through a multistep radioactivity-based approach. We have recently shown that sites of O-GlcNAc modification can be determined using quadrupole time-of-flight tandem mass spectrometry (Chalkley, R. J., and Burlingame, A. L. (2001) Identification of GlcNAcylation sites of peptides and alpha-crystallin using Q-TOF mass spectrometry. J. Am. Soc. Mass Spectrom. 12, 1106-1113). In this work utilization of this new approach has revealed previously undetected sites of O-GlcNAc modification of the transcription factor serum response factor.
Chalkley
Robert J
RJ
Department of Biochemistry and Molecular Biology, University College, London, Gower Street, London WC1E 6BT, United Kingdom. robertc@itsa.ucsf.edu
Burlingame
A L
AL
D000117:Acetylglucosamine / Q000737:chemistry*; D000595:Amino Acid Sequence; D002851:Chromatography, High Pressure Liquid; D004591:Electrophoresis, Polyacrylamide Gel; D006801:Humans; D008969:Molecular Sequence Data; D010766:Phosphorylation; D026362:Serum Response Factor / Q000737:chemistry*; D021241:Spectrometry, Mass, Electrospray Ionization
D026362:Serum Response Factor; D000117:Acetylglucosamine