Protein Sequencer/Amino Acid Analyzer

Cohn, David

Abstract

This application for a Shared Equipment Grant request funds to establish a protein sequencing facility. The users propose to purchase an Applied Biosystems Model 470A gas phase sequencer and a Waters Model 840 parallel-channel HPLC. The latter will be configured to simultaneously analyze PTH-amino acids arising from the Edman degradation sequencing procedure and amino acid compositions of acid hydrolysates of specific proteins and peptides. This equipment will provide a central facility for the analysis and composition of proteins and peptides to support ongoing research activities. The University of Louisville Graduate and Medical Schools have committed long-term funding for maintenance of the instruments and their operation. Research projects that depend on, or will benefit from, the sequencer facility include: analysis of peptide and proteins of the parathyroid gland involved in the processing and secretion of parathormone; study of membrane proteins associated with a human Ca++-pump and comparison to the skeletal and cardiac muscle Ca++-ATPase enzyme partial sequencing of peptides contained within human serum albumin involved in the specific binding of drugs and non-enzymatic glycosylation; characterization of peptides generated from mammalian tumor and synovial collagenases; study of peptides involved in the interactions of cytochrome P-450 and NADPH-P-450 enzymes (Gray); sequencing of S-adenosyl homocysteinase to an extent sufficient to prepare oligonucleotide probes for screening a cDNA library for the gene; a similar project to develop oligonucleotide probes to mammalian """"""""stress"""""""" protein genes; structural studies and intercomparisons of unique 34 kDa phosphorylated proteins of mammalian cancer cells related to the binding of epidermal growth factor; structure-activity studies of hormonally active peptides and peptide enzyme inhibitors; and chemical analysis of a new chemotactic factor derived from eosinophils.