Formation of angiotensin II and other angiotensin peptides from des-leu 10-angiotensin I in rat lung and kidney.
The formation of AII from a metabolite of AI, des-leu10-angiotensin I [A(1-9)] has been studied in centrifugal fractions of rat lung and kidney using gradient elution HPLC to monitor the formation of peptide products. AII-forming activity was present in kidney S2 (22.3 nmol/mg protein/min) but not in kidney P2 centrifugal fractions. Lung S2 fractions showed relatively weak AII-forming activity (0.34 nmole/mg protein/min) whilst no activity was observed in lung P2. Carboxypeptidase N-like activity measured using both Hipp-Arg and Hipp-Lys as synthetic substrates did not parallel AII-forming activity, since this activity was highest in the P2 fractions of both lung and kidney, as were ACE and aminopeptidase activities. Whilst the major peptide produced in kidney S2 was AII (71%) significant amounts of both AIII (23%) and A(2-9) (6%) were also observed. In lung the amounts of these peptides produced as a percentage of the A(1-9) degrading activity were 2.9%, 2.4% and 21% respectively. The AII-forming activity in kidney S2 was not inhibited by enalaprilat, bestatin, amastatin, phosphoramidon or Pro-Phe but was inhibited (31%) by 1 mM cobalt (II). 1,10-Phenanthroline, iodoacetic acid, EDTA and puromycin significantly enhanced the formation of AII and increased the rate of degradation of the substrate, A(1-9). These results support the concept of a sequential carboxypeptidase pathway operating, particularly in kidney, to produce AII from AI. These results provide further evidence of an alternative metabolic pathway for the formation of AII not involving angiotensin converting enzyme.
Drummer
O H
OH
University of Melbourne, Austin Hospital, Heidelberg, Victoria, Australia.
Kourtis
S
S
Johnson
H
H
D000626:Aminopeptidases / Q000378:metabolism; D000803:Angiotensin I / Q000031:analogs & derivatives* / Q000378:metabolism; D000804:Angiotensin II / Q000378:metabolism*; D000818:Animals; D002851:Chromatography, High Pressure Liquid; D007668:Kidney / Q000378:metabolism*; D008168:Lung / Q000378:metabolism*; D001121:Lysine Carboxypeptidase / Q000378:metabolism; D007703:Peptidyl-Dipeptidase A / Q000378:metabolism; D011480:Protease Inhibitors / Q000494:pharmacology; D051381:Rats; D011919:Rats, Inbred Strains
D011480:Protease Inhibitors; D000804:Angiotensin II; C045857:angiotensin I, des-Leu(10)-; D000803:Angiotensin I; D000626:Aminopeptidases; D007703:Peptidyl-Dipeptidase A; D001121:Lysine Carboxypeptidase