Degradation of proinsulin and isolated C-peptide by rat kidney neutral metallo-endopeptidase.
Previous studies have shown that a neutral metallo-endopeptidase purified from rat kidney degrades the B chain of insulin, glucagon, ACTH and, at a markedly slower rate, the A chain of insulin. In contrast the enzyme does not attack native insulin, oxytocin, vasopressin, ribonuclease, albumin or denatured hemoglobin. The current studies demonstrate that the neutral peptidase also degrades the isolated C-peptide of proinsulin and cleaves certain peptide bonds in and near the C-peptide moiety of native proinsulin. Time courses of the formation of fluorescamine-reactive material during digestion of proinsulin and isolated C-peptide with the peptidase were identical. However, structural analysis of the peptidase-digested proinsulin showed that the enzyme does not convert proinsulin to insulin but that the peptidase cleaves one bond, Tyr26-Thr27, in the B chain moiety and five bonds in the C-peptide moiety, producing four split proinsulins. One of the split proinsulins is des-octacosa-peptide (27-54) porcine proinsulin or des-tetracosapeptide (27-50) bovine proinsulin. Each is a derivative of the insulin molecule having an extension of nine residues (ten residues in the case of the derivative from bovine proinsulin) at the N-terminus of A chain and lacking four residues at the C-terminus of B chain. This two chain derivative retains full immunoreactivity with insulin antibodies and exhibits 2.4-times more biological activity (promotion of glycogenesis in primary cultured hepatocytes) than proinsulin and about two-thirds the activity of insulin.
D000595:Amino Acid Sequence; D000818:Animals; D002096:C-Peptide / Q000378:metabolism*; D055598:Chemical Phenomena; D002621:Chemistry; D002850:Chromatography, Gel; D010450:Endopeptidases / Q000378:metabolism*; D007668:Kidney / Q000201:enzymology*; D008666:Metalloendopeptidases; D010455:Peptides / Q000378:metabolism*; D011384:Proinsulin / Q000378:metabolism*; D011863:Radioimmunoassay; D051381:Rats; D013379:Substrate Specificity
D002096:C-Peptide; D010455:Peptides; D011384:Proinsulin; D010450:Endopeptidases; D008666:Metalloendopeptidases