Separations and tandem mass spectrometry for shotgun proteomics

ANYL 27

John R. Yates III, jyates@scripps.edu, Akira Motoyama, and Catherine Wong. Cell Biology, The Scripps Research Institute, 10550 North Torrey Pines Road, SR11, LaJolla, CA 92037
A component to understanding biological processes involves identifying the proteins expressed in cells as well as their modifications and the dynamics of processes. Several major technologies have benefited from large scale genome sequencing of organisms. The sequence data produced by these efforts can be used to interpret mass spectrometry data of proteins and thus enables rapid and straightforward analysis of protein data from experiments. Proteins separated by gel electrophoresis and 2-dimensional gel electrophoresis can now be rapidly identified enabling more comprehensive analyses of biochemical and molecular biology experiments. New approaches have also emerged for protein analysis such as direct identification of proteins in mixtures without gel separation. By digesting protein mixtures and separating peptides with liquid chromatography directly into tandem mass spectrometers, sufficient information can be obtained to identify the peptides and subsequently the proteins present in the mixture. As peptide mixtures become more complex better separation techniques such as 2-dimensional liquid chromatography are required to resolve the peptide components for analysis. New 2-D LC approaches for peptide separations will be discussed.