The metabolism of neuropeptides. Hydrolysis of peptides by the phosphoramidon-insensitive rat kidney enzyme 'endopeptidase-2' and by rat microvillar membranes.
Endopeptidase-2, the second endopeptidase in rat kidney brush border [Kenny & Ingram (1987) Biochem. J. 245, 515-524] has been further characterized in regard to its specificity and its contribution to the hydrolysis of peptides by microvillar membrane preparations. The peptide products were identified, after incubating luliberin, substance P, bradykinin and angiotensins I, II and III with the purified enzyme. The bonds hydrolysed were those involving a hydrophobic amino acid residue, but this residue could be located at either the P1 or P1' site. Luliberin was hydrolysed faster than other peptides tested, followed by substance P and bradykinin. Human alpha-atrial natriuretic peptide and the angiotensins were only slowly attacked. Oxytocin and [Arg8]vasopressin were not hydrolysed. No peptide fragments were detected on prolonged incubation with insulin, cytochrome c, ovalbumin and serum albumin. In comparison with pig endopeptidase-24.11 the rates for the susceptible peptides were, with the exception of luliberin, much lower for endopeptidase-2. Indeed, for bradykinin and substance P the ratio kcat./Km was two orders of magnitude lower. Since both endopeptidases are present in rat kidney microvilli, an assessment was made of the relative contributions to the hydrolysis of luliberin, bradykinin and substance P. Only for the first named was endopeptidase-2 the dominant enzyme; for bradykinin it made an equal, and for substance P a minor, contribution.
Stephenson
S L
SL
Department of Biochemistry, University of Leeds, U.K.
Kenny
A J
AJ
D000809:Angiotensins / Q000378:metabolism; D000818:Animals; D009320:Atrial Natriuretic Factor / Q000378:metabolism; D001920:Bradykinin / Q000378:metabolism; D002851:Chromatography, High Pressure Liquid; D006020:Glycopeptides / Q000494:pharmacology; D007987:Gonadotropin-Releasing Hormone / Q000378:metabolism; D006868:Hydrolysis; D007668:Kidney / Q000201:enzymology*; D007700:Kinetics; D008666:Metalloendopeptidases / Q000378:metabolism*; D008871:Microvilli / Q000201:enzymology; D015260:Neprilysin / Q000378:metabolism; D009479:Neuropeptides / Q000378:metabolism*; D051381:Rats; D013373:Substance P / Q000378:metabolism; D013552:Swine
D000809:Angiotensins; D006020:Glycopeptides; D009479:Neuropeptides; D013373:Substance P; D007987:Gonadotropin-Releasing Hormone; D009320:Atrial Natriuretic Factor; D008666:Metalloendopeptidases; D015260:Neprilysin; C053833:meprin A; D001920:Bradykinin; C008890:phosphoramidon