This application requests an Applied Biosystems model 477A/120A automated Protein Sequencer and a free-standing microbore HPLC (ABI model 130A) for interdepartmental use by investigators in the School of Medicine and Biomedical Sciences of the State University of New York at Buffalo. 'Mere is no protein sequencer in the School of Medicine. Nor is sequencing available routinely on a fee-for-service basis in the University at Buffalo, that can satisfy the needs of this user group. When samples are accepted, they are accepted on a """"""""second"""""""" priority basis. Four applicants are having sequencing done outside of the University, and others are planning to do the same. The model 477A is requested, rather than the less costly model 473A, because of its 2-fold advantage in sensitivity and its greater versatility. The microbore HPLC will expedite optimum use of the sequencer and will enhance the sequencing facility's utility. The sequencer will be operated and maintained by a skilled technician with previous experience in operating an automated protein sequencer. The HPLC will be operated by individual users under the supervision of the PI. The technician's salary will be derived from state funds through the Department of Biochemistry, major repairs will be offset by the School of Medicine, other repairs and some operating costs will be provided by the Center for Applied Molecular Biology and Immunology, and the remaining routine operating costs will be obtained through user fees. A three-person internal advisory committee, consisting of the PI and two of the Co-PIs who have the most pressing need for this instrument, will govern the care and use of the instrument. The instrument will serve a core user group of 14 investigators, representing 18 projects, all supported by the National Institutes of Health, with the exception of one, who is supported by the NSF. The instruments will be in use about 90% of the time. The projects can be grouped into two categories, with several fitting into both. The first type concentrates on protein chemistry and enzymology. These users require protein/peptide sequencing routinely to derive functional information, such as the nature of active sites, precursor product relationships, or confirmation of sequences in site-directed mutagenized proteins. Six of the applicants have these needs (projects 1 through 5 and 13). The second type will use sequence information from tryptic digests of partially-purified enzymes/proteins to plan syntheses of oligopeptides and oligonucleotide probes for raising antibodies and cloning the cDNA of the genes of the target proteins. Ten of the applicants have this need (projects 1 and 6 through 18). A protein sequencer and dedicated microbore HPLC will enhance funded research by providing the capacity and accessibility required for optimum progress. An additional advantage is that the cost to the individual will be lower than other alternatives.
