In this proposal, we are requesting support for the purchase of an instrument system to semi-automate sample processing for proteomic projects performed in the University of Virginia Biomolecular Research Facility for University investigators. Specifically, we are requesting a gel imaging system and associated software, gel spot picker, gel spot digester and digest spotter. Samples generated by these instruments will be subjected to automated mass mapping on our existing PE Voyager DE Pro MALDI-TOF instrument or if necessary, de novo sequencing with the facility's Finnigan LCQ ion-trap LC/MS/MS mass spectrometer. The overall goal of this program is to provide high throughput, low cost protein identification for investigators with proteomic related projects. Furthermore, this technology will be interfaced with the Microarray/GeneChip Program in the facility. The instruments will be housed in the University of Virginia Biomolecular Research Facility and will support the research proteomic projects of seven NIH-funded investigators. For these projects, we anticipate that the instruments will be in use approximately 80% of the available time. The remaining time will be available for new proteomic-related project development. There is no other mass spectrometric protein identification service, high throughput or otherwise, available at the University. Dr. Jay Fox, Director of the Biomolecular Research Facility, will be responsible for the overall management of the instrumentation. Dr. Nicholas Sherman will be responsible for mass spectrometry issues of proteomics projects and Dr. John Shannon will be responsible for the protein chemistry aspects of the proteomic projects. Both are staff members in the Biomolecular Research Facility. All three scientists will consult with investigators in the planning, execution and data analysis of proteomics projects. For the past seven years, the facility has offered mass spectrometric protein identification and de novo sequence analysis and no major difficulties are expected on the downstream end of sample analysis. A fee-for-service charge will be instituted to help defray the costs of staff salaries and supplies. Oversight of the Biomolecular Research Facility is provided by a standing committee appointed by the Dean of the Medical School. The Oversight Committee provides guidance and expertise directly to Dr. Fox for the management of the facility as well as complementing the scientific expertise of facility staff. One of the principal investigators in this proposal, Dr. Kevin Hazen, is a member of the committee. The University of Virginia Biomolecular Research Facility has been in operation for the past 20 years with continual fiscal support from the University of Virginia Health Sciences Center. The administration of the Health Sciences Center is committed to the long-term operation of the facility. This support, along with that of the Oversight Committee and facility staff, will serve to ensure adequate, appropriate instrument use for the fiscally sound and scientifically successful operation of instrumentation requested for the benefit of NIH-funded investigators at the University of Virginia..
