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Molecular & Cellular Proteomics 2:182-190, 2003.
© 2003 by The American Society for Biochemistry and Molecular Biology, Inc.

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Research

Identification of Novel Sites of O-N-Acetylglucosamine Modification of Serum Response Factor Using Quadrupole Time-of-flight Mass Spectrometry^*

Robert J. Chalkley and A. L. Burlingame

From the Department of Biochemistry and Molecular Biology, University College, London, Gower Street, London WC1E 6BT, United Kingdom and the Ludwig Institute for Cancer Research, 91 Riding House Street, London W1W 7BS, United Kingdom

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 -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.

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