Proteomics and Molecular Characterization

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PROTEOMICS SHARED RESOURCE The overall goal of the Proteomics and Molecular Characterization Shared Resource is to provide a wide range of state-of-the-art protein analysis services for KCC investigators. These services are grouped into three major areas: 1) Proteomics and Mass Spectrometry performs experiments designed to help investigators identify and characterize proteins on a global scale (e.g. expression levels, protein complexes, post-translational modifications). Protein identification is accomplished with mass spectrometry (SELDI and MALDI TOP and LC ESI). Currently, 2-Dimensional PAGE analysis is the predominant platform used to define differences in protein expression between experimental samples, but we have made significant efforts to expand the services to include multidimensional chromatography-based approaches. In our estimation, this will be the preferred workflow for many investigators especially those involved with clinical studies because the HPLC analysis requires significantly less starting material and because of less variability compared with the gel based separations. Commercial kits for isotopic labeling (e.g. SILAC, ICAT and ITRAQ) as well as 18O labeling of tryptic peptides are being developed for KCC members. 2) Protein and Peptide Chemistry provides peptide synthesis services for KCC investigators at very reasonable costs. In addition to routine peptides, the resource has successfully synthesized many specialized peptidesThe resource has been successful in synthesizing extremely long peptides (up to 69 residues), as well as cyclic peptides. 3) Molecular Interaction provides access to instrumentation for characterizing protein/protein interactions. These instruments include a Biacore 3000 SPR biosensor, Kinexa 3000, and an Isothermal Titration Calorimeter. In addition to the services contained within these Divisions, the shared resource provides users access to a SELDI-TOF mass spectrometer, a MALDI-TOF mass spectrometer, Typhoon 9400 Gel Imager, UV spectrometer, as well as equipment for sonication, vacuum centrifugation, and lyophilization. The services and equipment made available by this shared resource play an important role in the current and future research plans of KCC investigators. This shared resource underwent significant management changes in December 2006 to improve the resource by integrating two active research laboratories whose research is focused on proteomics and mass spectrometry. Our goals are threefold: 1) to increase the number of services available to researchers; 2) to reduce costs and increase chargebacks; and 3) to bring new instrumentation to the resource. Additionally, usage is expected to increase as the resource begins offering new services based on LC/MS proteomics workflows and protein microarrays. These changes will improve the capabilities while decreasing the overall costs of running the Facility.