Modular mass spectrometric tool for analysis of composition and phosphorylation of protein complexes.
The combination of high accuracy, sensitivity and speed of single and multiple-stage mass spectrometric analyses enables the collection of comprehensive sets of data containing detailed information about complex biological samples. To achieve these properties, we combined two high-performance matrix-assisted laser desorption ionization mass analyzers in one modular mass spectrometric tool, and applied this tool for dissecting the composition and post-translational modifications of protein complexes. As an example of this approach, we here present studies of the Saccharomyces cerevisiae anaphase-promoting complexes (APC) and elucidation of phosphorylation sites on its components. In general, the modular concept we describe could be useful for assembling mass spectrometers operating with both matrix-assisted laser desorption ionization (MALDI) and electrospray ionization (ESI) ion sources into powerful mass spectrometric tools for the comprehensive analysis of complex biological samples.
Blethrow
Justin D
JD
Department of Cellular and Molecular Pharmacology, University of California San Francisco, San Francisco, California, United States of America.
Tang
Chao
C
Deng
Changhui
C
Krutchinsky
Andrew N
AN
D000595:Amino Acid Sequence; D008969:Molecular Sequence Data; D010766:Phosphorylation; D029701:Saccharomyces cerevisiae Proteins / Q000737:chemistry / Q000378:metabolism*; D021241:Spectrometry, Mass, Electrospray Ionization / Q000379:methods*; D019032:Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization / Q000379:methods*
D029701:Saccharomyces cerevisiae Proteins