The purpose of this proposal is to seek funds for the purchase of a semi-robotic Proteome Works system. This integrated instruments and software package will greatly enhance the Laboratory of Mass Spectrometry with the Lerner Research Institute of the Cleveland Clinic Foundation (CCF) capabilities. The requested integrated protein spot cutter, protein proteolytic digestion/processing/MALDI-spotting station and software will be used to facilitate and advance research programs at institutions within the area (CCF and Case Western Reserve University). Major Users all have numerous NIH-funded research grants, which have component projects that can best be conducted using 2-D gel analysis followed by mass spectrometry. All the Major User's projects require substantial numbers of sample preparations and/or 2-D gel analysis and comparison. The Major Users projects are broad and include: 1) identification of differentially expressed proteins in normal and diseased tissue; 2) quantification of structurally defined components in complex biological complexes; 3) characterization of covalent protein modifications; and 4) investigations of cellular differentiation markers and altered gene expression. The proteome Works system provides the versatility, resolution and capabilities to meet these diverse demands. In addition, the requested instrument system will be used to meet the needs of the occasional user. This service will be conducted within our existing Molecular Biotechnology Core, which provides support to investigators by performing protein sequencing, peptide synthesis, and related services. Due to the human genome project, the Mass Spectrometry Core has experienced greatly increased usage. With the integrated Proteome Works system, the mass Spectrometry Core can process more samples and increase its machine-time efficiency. Importantly, the Proteome Works system will allow users to digitally capture 2-D gel images, compare results of multiple experiments and integrate in gene expression information and relate it directly back to protein identities and expression levels. In fact, the inability to do this has significantly hindered the pace of progress in biomedical research. Acquisition of the Proteome Works system will allow full utilization of the unique capabilities and usefulness of the Mass Spectrometry Core, gene expression laboratories and Cores here at CCF and CWRU. CCF has committed substantial resources to the development of the CCF Laboratory of Mass Spectrometry, including personnel to oversee its operation, and a fiscal plan to insure its operation and maintenance for the long-term. In sum, acquisition of the requested integrated Proteome Works system will have a significant impact on the progress and future development of biomedical research programs at multiple institutions in the region including CCF and CWRU.
