Mechanism of digestion of bradykinin and lysylbradykinin (kallidin) in human serum. Role of carboxypeptidase, angiotensin-converting enzyme and determination of final degradation products.
Degradation of bradykinin and lysylbradykinin was studied in plasma and serum, and the results were compared to those seen with mixtures of carboxypeptidase N and angiotensin-converting enzyme (ACE), the two recognized kininases in blood. Angiotensin-converting enzyme was an effective kininase in mixtures with carboxypeptidase N at physiologic concentration and digested bradykinin to the dipeptides Phe- Arg and Ser-Pro plus the pentapeptide Arg-Pro-Pro-Gly-Phe. Carboxypeptidase N slowly removed the C-terminal Arg from bradykinin to yield des-Arg9-bradykinin (DBK); the latter was digested by ACE to yield the aforementioned pentapeptide and the tripeptide Ser-Pro-Phe. In serum, however, the C-terminal Arg was removed from bradykinin about five times faster than was accounted for by the activity of carboxypeptidase N. The primary substrate of ACE in serum, therefore, was des-Arg9-bradykinin and not bradykinin. The products of this reaction, pentapeptide and tripeptide, were unstable in serum and were cleaved by enzymes that have not yet been characterized. One product, free phenylalanine, was used to monitor these reactions by HPLC. Our studies indicate that the final products of bradykinin degradation were the tripeptide Arg-Pro-Pro, one mole each of Ser, Pro, Gly, and Arg, and two moles of phenylalanine. Since the serum level of carboxypeptidase N did not account for the rapid kinin degradation seen, other carboxypeptidases may have been operative, perhaps released as a result of blood clotting, or a serum cofactor may augment carboxypeptidase N activity.
Sheikh
I A
IA
Department of Medicine, SUNY-Stony Brook 11794-8160.
Kaplan
A P
AP
D000818:Animals; D001920:Bradykinin / Q000097:blood*; D002268:Carboxypeptidases / Q000097:blood*; D002851:Chromatography, High Pressure Liquid; D006801:Humans; D006868:Hydrolysis; D007201:Indicator Dilution Techniques; D007609:Kallidin / Q000097:blood*; D010446:Peptide Fragments / Q000097:blood*; D007703:Peptidyl-Dipeptidase A / Q000097:blood*
D010446:Peptide Fragments; D007609:Kallidin; D002268:Carboxypeptidases; D007703:Peptidyl-Dipeptidase A; D001920:Bradykinin