APPLIED BIOSYSTEMS MODEL 420A-03 DERIVATIZER-ANALYZER

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This proposal is being submitted in order to obtain an Applied Biosystems 420A Derivatizer-Analyzer system for highly sensitive amino acid analysis. Project I consists of the structural characterization of several proteins involved with the complement system. Dysfunctional C1 inhibitor proteins from patients with hereditary angioneurotic edema will be analyzed to find the structural determinants of dysfunction. The kinin-like peptide also present in these patients' plasma will be isolated and characterized. The structure of biologically active fragments of C3 will be determined, as will the structure of bovine conglutinin. Project II is involved with the structural and functional characterization of bone phosphoproteins, and of enamel proteins and phosphoproteins. Project III is designed to specifically identify the region of IgG in immune complexes to which complement component C4 covalently binds during complement activation. C4 isotypes will be compared in their binding to immune complexes. Project IV is concerned with characterization of T cell control of IgE synthesis. The T cell IgE receptor will be isolated from T cells of patients with the hyper-IgE syndrome. Project V is involved with calcium-binding proteins and other proteins of the mineralized extracellular matrix in bone. Osteocalcin fragments present in bone matrix will be characterized. The activation of tumor cell procathepsin B will be analyzed, and the structure and function of polypeptide growth factors from bone matrix will be characterized. Project VI is directed toward the characterization of receptors and ion channels from cardiac and skeletal muscle. The calcium channel from fast skeletal muscle sarcolemmel membranes and the fast calcium release channel from the terminal cisternae of the sarcoplasmic reticulum are being isolated; their amino acid compositions will be determined and partial sequence analysis done in order to clone the genes for the proteins.