title: Endoproteolysis at tetrabasic amino acid sites in procalcitonin gene-related peptide by pituitary cell lines.
abstract: The specificity of neuroendocrine prohormone convertases for tetrabasic amino acid sites was investigated. Mutations were introduced into the tetrabasic cleavage site of the procalcitonin gene-related peptide (proCGRP) cDNA and these mutated cDNA's were expressed in AtT-20 cells which predominantly express the endoprotease prohormone convertase-1 (PC1/3), and in GH3 cells which predominantly express prohormone convertase-2 (PC2). Mutations were introduced into the proCGRP cDNA which converted the wild-type ArgArgArgArg site to LysLysArgArg and ArgArgLysLys, and the proCGRP variants were stably transfected into AtT-20 and GH3 cells. ProCGRP containing each of the LysLysArgArg permutations were efficiently cleaved in both AtT-20 and GH3 cells. Cleavage of LysLysArgArg in exogenous proCGRP, but not in endogenous POMC, suggests that the specificity of cleavage at tetrabasic sites is not defined solely by the endoproteases expressed by the cell or by the amino acid sequence at the cleavage site, but is also dependent on the structure of the propeptide.
authors.affiliation: Department of Biochemistry and Molecular Biology, University of Miami School of Medicine, FL 33101, USA.
authors.forename: M I
authors.initials: MI
authors.lastname: Rosenblatt
authors.forename: I M
authors.initials: IM
authors.lastname: Dickerson
mesh_terms: D000596:Amino Acids / Q000737:chemistry*; D016282:Aspartic Acid Endopeptidases / Q000378:metabolism*; D001665:Binding Sites; D015740:Calcitonin Gene-Related Peptide / Q000378:metabolism*; D002460:Cell Line; D016297:Mutagenesis, Site-Directed; D010902:Pituitary Gland / Q000166:cytology / Q000378:metabolism*; D045706:Proprotein Convertase 2; D043484:Proprotein Convertases; D011498:Protein Precursors / Q000378:metabolism*; D013379:Substrate Specificity; D013381:Subtilisins / Q000378:metabolism*
chemical_list: D000596:Amino Acids; D011498:Protein Precursors; C075733:pro-calcitonin gene-related peptide; D043484:Proprotein Convertases; D013381:Subtilisins; D045706:Proprotein Convertase 2; D016282:Aspartic Acid Endopeptidases; D015740:Calcitonin Gene-Related Peptide