This Shared Instrumentation Grant proposal is for the purchase of the ProteomeWorks(TM) System from BIO-RAD and Micromass. This is an integrated system for proteomics studies consisting of 1) 2-D gel electrophoresis equipment, gel image and analysis equipment, and a 2-D gel spot cutter from BIO-RAD; and 2) the MassPREP Station, TofSpec- 2E MALDI-TOF mass spectrometer, and software for database searches from Micromass. This proteomics system will be operated out of the Baylor College of Medicine (BCM) Protein Chemistry Core Laboratory and supported by the Office of Research. A proteomics core does not exist at BCM nor is similar instrumentation available. The College simply does not have the capacity to perform the high throughput level of identification of novel proteins, interacting proteins, o important posttranslational modifications needed by the following investigators. A total of 12 users from departments are included in this application. Users, and their projects, who will share the 2-D gel/MALDI-TOF system for proteomics studies include: 1) Joseph Bryan, ATP-sensitive K+ channels; 2) Stephen Elledge, the role of cell cycle genes in development and cancer; 3) Susan Hamilton, activity dependent signal transduction in skeletal muscle; 4) Wade Harper, proteomics approaches to cell cycle control; 5) Mary Moore, 2-D gel electrophoresis and MS analysis in the study of the nuclear transport machinery; 6) Bert O'Malley, mechanisms of nuclear receptor action in normal and malignant tissues; 7) Jun Qin, identification of phosphopeptides and proteins in BRCA and ATM protein complexes; 8) David Roth, identification of novel components of the V(D)J recombinase complex; 9) Robert Schwartz, a strategy to identify additional interactive SRF accessory factors; 10) David Sweatt, protein synthesis alterations in mammalian associative learning; 11) Nancy Weigel, steroid receptor phosphorylation and regulation of prostate cancer growth by vitamin D; and 12) Theodore Wensel, proteomics of vertebrate phototransduction. For all of these projects, the requested 2-D gel/MALDI-TOF ProteomeWorks(TM) System is essential for accomplishing the stated goals.
