IMPROVED METHODS FOR THE ANALYSIS OF PROTEIN COMPLEXES

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This subproject is one of many research subprojects utilizing the resources provided by a Center grant funded by NIH/NCRR. The subproject and investigator (PI) may have received primary funding from another NIH source, and thus could be represented in other CRISP entries. The institution listed is for the Center, which is not necessarily the institution for the investigator. We have juxtaposed several methods in an effort to improve the identification of proteins in complex protein mixtures (5-50 proteins.). Thus, we have compared direct digestion of protein mixtures followed by tryptic peptide liquid chromatography-mass spectrometric identification of peptides in chromatographic fractions with in-gel separation of proteins followed by complete gel slicing and mass spectrometric identification of tryptic peptides in every piece of a gel slice. Optimization of our total gel slicing technique is leading to an improved method for solving challenging biological problems described in the Collaborative Research section.