This Shared Instrumentation Grant proposal is for the purchase of equipment to be used in the isolation of proteins and peptides and the primary structural analysis of proteins/peptides. The requested items include the Applied Biosystems, Inc. (ABI) Model 477-A Pulsed Liquid Phase Protein Sequencing System, the ABI Model 420-A Automated Amino Acid Analyzer, and an accessory item for protein isolation, the ABI Model 230-A High Performance Electrophoresis System (HPEC). This instrumentation represents necessary additions to the Baylor College of Medicine Protein Chemistry Core Facility. A total of 18 users from 9 different departments at the College are included in this proposal. Investigators who will share this instrumentation and their project areas include: D. Allis (nuclear proteins and enzymes from Tetrahymena); K. Angelides (ion channel proteins and neurofilaments); M. Birnbaumer (vasopressin receptors); J. Butel (SV40 T antigen associated proteins); B. Dunbar (ovarian glycoproteins); H. Epstein (thick filaments and cardiac muscle proteins); S. Eskin (endothelial cell shear stress induced proteins); M. Estes (rotavirus proteins); S. Hamilton (calcium channel proteins from muscle); F. Ledley (cobalamin dependent metabolic enzymes); E. McCabe (metabolic enzymes glycerol kinase and hexokinase); D. Marcus (anti-carbohydrate antibodies); T. Means (calmodulin and calmodulin dependent enzymes); D. Medina (selenium binding proteins); B. O'Malley (steroid receptors and transcription factors); B. Perryman (creatine kinase from plasma); S. Rich (immunoregulatory lymphokines and factors); S. Tsai (ovalbumin gene transcription factors). The equipment requested will be used to sequence these biologically important proteins and peptides, to sequence on synthetic peptides and on isolated functionally important proteins/peptides, and to perform rapid electrophoretic isolations of proteins/peptides. Primary structural data generated with this instrumentation will be essential to progress of each of the biomedical research projects described in this proposal. The protein sequencer, amino acid analyzer, and automated electrophoresis system represent state-of-the- art instruments that are required additions to an existing, functional, and successful Protein Microchemistry Facility that is currently heavily utilized. A present and future constraint on the Facility is the lack of """"""""instrument time"""""""" for the multiple users at the College. The requested instrumentation will help to alleviate this problem and provide for more rapid and timely analyses.
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