Carboxypeptidase E is a sorting receptor for prohormones: binding and kinetic studies.
The binding of pro-opiomelanocortin,(POMC), pro-insulin, pro-enkephalin and chromogranin A (CGA) to the regulated secretory pathway sorting receptor, carboxypeptidase E (CPE), in bovine pituitary secretory granule (SG) membranes was investigated. N-POMC1-26, which contains the POMC sorting signal, bound to CPE in the SG membranes with low affinity and the binding was ion independent. Pro-insulin bound CPE with similar kinetics. Pro-enkephalin, but not CGA bound to CPE with similar IC50 as pro-insulin and N-POMC1-26. Crosslinking studies showed that pro-insulin and pro-enkephalin bound specifically to SG membrane CPE, similar to N-POMC1-26 reported previously. CPE was extracted from the SG membranes with NaHCO3 or KSCN, but not Triton X-100/1 M NaCl. The results show that CPE is tightly associated with SG membranes and binds several prohormones, but not CGA, with similar kinetics, providing further evidence that membrane CPE has the characteristics to function as a common sorting receptor for targeting prohormones to the regulated secretory pathway.
Section on Cellular Neurobiology, Laboratory of Developmental Neurobiology, National Institute of Child Health and Human Development, National Institutes of Health, Bethesda, MD 20892, USA.
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D000818:Animals; D002118:Calcium; D043423:Carboxypeptidase H; D002268:Carboxypeptidases / Q000737:chemistry* / Q000378:metabolism; D002417:Cattle; D053379:Chromogranin A; D002864:Chromogranins / Q000737:chemistry / Q000378:metabolism; D003432:Cross-Linking Reagents; D003594:Cytoplasmic Granules / Q000201:enzymology*; D007425:Intracellular Membranes / Q000201:enzymology; D007700:Kinetics; D017830:Octoxynol; D010902:Pituitary Gland / Q000201:enzymology; D011498:Protein Precursors / Q000737:chemistry / Q000378:metabolism; D011956:Receptors, Cell Surface / Q000737:chemistry* / Q000378:metabolism
D053379:Chromogranin A; D002864:Chromogranins; D003432:Cross-Linking Reagents; D011498:Protein Precursors; D011956:Receptors, Cell Surface; D017830:Octoxynol; D002268:Carboxypeptidases; D043423:Carboxypeptidase H; D002118:Calcium