Regulation of angiotensin I-converting enzyme in cultured bovine bronchial epithelial cells.
The purpose of this study was to determine whether angiotensin I-converting enzyme (ACE) is present in cultured bovine bronchial epithelial cells (BBECs) and whether its activity can be modulated. We found that extracts of confluent monolayers of cultured BBECs degraded [glycine-1-14C]hippuryl-L-histidyl-L-leucine at a rate of 843 +/- 66 pmol/hr/mg protein (mean +/- SEM, n = 5). In addition, we found that the enzyme was shed into the culture medium. ACE activity in BBECs was inhibited by three selective, but structurally different, ACE inhibitors (captopril, quinapril, and cisalaprilat) with an IC50 of approximately 2 nM. Increasing chloride concentration in the assay buffer resulted in an increase in BBECs ACE activity of 63%. Enzyme activity was also modulated by the presence of zinc cation in the assay buffer. Addition of dexamethasone to the culture medium was associated with a significant increase in BBECs ACE activity (P < 0.05), which was inhibited by the steroid receptor antagonist RU 38486. Western blot analysis of BBECs, tracheal and bronchial mucosal strips utilizing a cross-reacting rabbit anti-mouse ACE antibody, showed a faint 175 kDa band and additional strong 52 kDa and 47 kDa band. The mechanism of generation of the low M.W. bands is unknown. Our data indicate the presence of ACE in cultured BBECs and that enzyme activity can be modulated.
Müns
G
G
Department of Internal Medicine, University of Nebraska Medical Center, Omaha 68198.
Vishwanatha
J K
JK
Rubinstein
I
I
D000806:Angiotensin-Converting Enzyme Inhibitors / Q000494:pharmacology; D000818:Animals; D000838:Anions; D015153:Blotting, Western; D001980:Bronchi / Q000201:enzymology*; D002417:Cattle; D002478:Cells, Cultured; D002712:Chlorides / Q000494:pharmacology; D003907:Dexamethasone / Q000494:pharmacology; D004848:Epithelium / Q000201:enzymology; D015735:Mifepristone / Q000494:pharmacology; D007703:Peptidyl-Dipeptidase A / Q000378:metabolism*; D015032:Zinc / Q000494:pharmacology
D000806:Angiotensin-Converting Enzyme Inhibitors; D000838:Anions; D002712:Chlorides; D015735:Mifepristone; D003907:Dexamethasone; D007703:Peptidyl-Dipeptidase A; D015032:Zinc