title: Proteolytic processing of presecretory proteins is required for development of biological activities in pancreatic exocrine proteins.
abstract: The biological activities of pancreatic presecretory and secretory proteins synthesized in vitro were compared in studies of (a) the binding of nascent amylase to its substrate, glycogen, (b) the binding of nascent trypsinogen 1, trypsinogen 2+3, and chymotrypsinogen 1 to Sepharose-bound soybean trypsin inhibitor, and (c) the activation of nascent trypsinogen by porcine enterokinase. Nascent secretory proteins synthesized in vitro using a mRNA-dependent gel-filtered reticulocyte lysate translation system supplemented with canine pancreas rough microsomes or canine pancreas mRNA and micrococcal nuclease-treated microsomal membranes showed biological activities similar to authentic secretory proteins if oxidized glutathione was added during their synthesis. Proteins synthesized in the presence of membranes and the absence of glutathione showed significantly less biological activity due to incorrect development of conformation. Presecretory proteins synthesized in vitro with canine pancreas mRNA in the absence of microsomal membranes had little or no activity after translation in either the absence or presence of glutathione. These and previous findings (Scheele, G. A., and Jacoby, R. (1982) J. Biol. Chem. 257, 12277-12282) indicate that proteolytic removal of the NH2-terminal transport peptide is necessary to allow correct conformational development, including the formation of native disulfide bonds, which not only stabilizes the molecule but allows expression of authentic biological and probiological activity.
mesh_terms: D000818:Animals; D002352:Carrier Proteins / Q000235:genetics* / Q000378:metabolism; D002919:Chymotrypsinogen / Q000235:genetics; D004285:Dogs; D010450:Endopeptidases / Q000235:genetics; D006003:Glycogen / Q000378:metabolism; D066298:In Vitro Techniques; D008861:Microsomes / Q000378:metabolism; D010179:Pancreas / Q000378:metabolism*; D011485:Protein Binding; D014176:Protein Biosynthesis; D012333:RNA, Messenger / Q000235:genetics; D012697:Serine Endopeptidases; D014362:Trypsinogen / Q000235:genetics
chemical_list: D002352:Carrier Proteins; D012333:RNA, Messenger; D014362:Trypsinogen; D006003:Glycogen; D002919:Chymotrypsinogen; D010450:Endopeptidases; D012697:Serine Endopeptidases