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Originally published In Press as doi:10.1074/mcp.M400041-MCP200 on June 15, 2004.
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Molecular & Cellular Proteomics 3:872-886, 2004.
© 2004 by The American Society for Biochemistry and Molecular Biology, Inc.

Research

Characterization of Tetrahymena Histone H2B Variants and Posttranslational Populations by Electron Capture Dissociation (ECD) Fourier Transform Ion Cyclotron Mass Spectrometry (FT-ICR MS)*,S

K. F. Medzihradszky{ddagger}, X. Zhang{ddagger}, R. J. Chalkley{ddagger}, S. Guan{ddagger}, M. A. McFarland§, M. J. Chalmers§, A. G. Marshall§, R. L. Diaz, C. D. Allis and A. L. Burlingame{ddagger},||

From the {ddagger} Department of Pharmaceutical Chemistry and Mass Spectrometry Facility, University of California, San Francisco, CA 94143-0446; § Ion Cyclotron Resonance Program, National High Magnetic Field Laboratory, Florida State University, Tallahassee, FL 32310-3706; and Laboratory of Chromatin Biology, Rockefeller University, New York, NY 10021

This work describes the nature and sequence information content of the electron capture dissociation mass spectra for the intact Tetrahymena histone H2B. Two major variants of this protein were present bearing nominal modifications of both +42 and +84 Da. This work describes identification of the nature of these two modifications. For example, using gas-phase selection and isolation of the +42-Da modified species, from a background of two H2B variants each present in six or more posttranslationally modified isoforms, we were able to determine that this +42-Da modification isoform bears trimethylation rather than acetylation. LC-CIDMS analysis was also employed on digested preparations to obtain complementary detail of the nature of site-specific posttranslational modifications. This study establishes that integration of the information from these two datasets provides a comprehensive map of posttranslational occupancy for each particular covalent assemblage selected for structural investigation.

|| To whom correspondence should be addressed: Department of Pharmaceutical Chemistry, University of California, 513 Parnassus Avenue, San Francisco, CA 94143-0446. Tel.: 415-476-5641; Fax: 415-476-0688; E-mail: alb{at}itsa.ucsf.edu


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