Degradation of bradykinin by bovine tracheal epithelium and isolated epithelial cells.
1. The degradation of bradykinin (BK) labelled with tritiated proline at positions 2 and 3 ([3H]-BK) was determined on the luminal surface of bovine tracheal epithelium, in supernatants obtained from incubations of the luminal tracheal surface, and in suspensions of isolated tracheal epithelial cells. Peptidase inhibitors and identification of peptide fragments were used for characterization of the metabolic pathways. 2. On the luminal surface of intact bovine trachea, [3H]-BK was degraded with a half life of 12.8 min. [1-7]-BK and [1-5]-BK were the major direct metabolites which were further degraded via [1-3]-BK and [2-3]-BK to proline. Metabolism of [3H]-BK was unaltered in the presence of ramiprilat (250 nM) or phosphoramidon (10 microM). Phenanthroline diminished the formation of [1-7]- and [1-5]-BK and abolished the generation of proline. 3. Supernatants obtained from incubations of tracheal epithelium contained kininase activities which steadily increased when tracheae were incubated for longer than 30 min. After 60 min contact with epithelium, the incubation medium contained higher kininase activities than the epithelium itself. The spectrum of kinin metabolites generated by kininases in the supernatant was comparable to that formed by intact epithelium. 4. In suspensions of isolated epithelial cells, [3H]-BK was degraded with a half life of 70 min. The metabolites [1-3]- and [2-3]-BK were formed in parallel to [1-7]- and [1-5]-BK; however, proline was not generated. Degradation of [3H]-BK was not influenced by ramiprilat, but was inhibited by 85% in the presence of phosphoramidon. Phosporamidon markedly inhibited the generation of [1-7]- and [1-5]-BK and nearly abolished the formation of [1-3]- and [2-3]-BK. 5. In conclusion, angiotensin I-converting enzyme and neutral endopeptidase 24.11 are not significantly involved in [3H]-BK degradation on the luminal side of intact tracheal epithelium. The spectrum of metabolites found may in fact reflect the combined activities of metalloendopeptidase 24.15 and post-proline cleaving enzymes. Enzymes showing similar kininase activities are also released from the epithelium. Isolated epithelial cells contain low activities of these kininases, but a high activity of neutral endopeptidases, which may reflect an exclusively basolateral localization of the latter.
Dendorfer
A
A
Institute of Pharmacology, Medical University of Lübeck, Germany.
Vordermark
D
D
Dominiak
P
P
D000818:Animals; D001920:Bradykinin / Q000378:metabolism*; D002417:Cattle; D004847:Epithelial Cells; D004848:Epithelium / Q000378:metabolism; D006207:Half-Life; D066298:In Vitro Techniques; D007705:Kinins / Q000378:metabolism; D008666:Metalloendopeptidases / Q000378:metabolism; D009479:Neuropeptides / Q000378:metabolism; D000085822:Prolyl Oligopeptidases; D012697:Serine Endopeptidases / Q000378:metabolism; D014132:Trachea / Q000201:enzymology / Q000378:metabolism*
D007705:Kinins; D009479:Neuropeptides; D012697:Serine Endopeptidases; D000085822:Prolyl Oligopeptidases; D008666:Metalloendopeptidases; C022510:thimet oligopeptidase; D001920:Bradykinin