In this subcontract analytical core of the program project entitled """"""""Innate immune response to microbial infection"""""""" we will provide expert mass spectrometric, chemical and chromatographic support. The analytical core will consist of chemistry, chromatography and mass spectrometry support. The analytical core will consist of chemistry, chromatography and mass spectrometry support. At UCSF, we have pioneered efforts in the structural determination and mass spectrometric analysis of bacterial lipooligosaccharides (LOS), oligosaccharides and lipid A. Our laboratory also has extensive experience in the application of mass spectrometric and chromatographic methods for the analysis of protein phosphorylation. In addition, we have recently carried out a successful proteomics investigation using 2-dimensional SDS-PAGE, mass spectrometry and database searching technologies. These technologies will serve as the basis of the analytical core support of the three projects that comprise this current program project application. For project #1 we will analyze and/or structurally characterize Neisseria NOS and lipid A species and identify proteins that are differentially expressed between wild-type and several mutant neisserial strains (e.g., htrb-). In project #2 we provide some structural characterization of glycolipids primarily via GC/MS and molecular weight profiling using mass spectrometry techniques. In the third and last project we will determine the phosphorylated state(s) of the key oxidative human enzyme p47-phox, using peptide mapping and mass spectrometry protocols. The structural studies that comprise the analytical core will provide important and essential data that will be key to obtaining a better understanding of the human response to bacterial infection.
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