Elevation of angiotensin converting enzyme in bovine endothelial cells quantitated by an ELISA.
Levels of angiotensin converting enzyme (ACE) in cultured bovine pulmonary artery endothelial cells treated with dexamethasone, aldosterone, 3,3',5'-triiodo-L-thyronine, Ca2+ ionophore, 3-isobutyl-1-methylxanthine, dibutyryl cAMP and forskolin were quantitated by an enzyme linked immunosorbent assay (ELISA). The configuration for the ELISA consisted of purified bovine lung ACE adsorbed to a solid phase competing with endothelial cellular ACE for a limited amount of anti-ACE immunoglobulin. ACE-IgG complex on the solid phase was detected by goat anti-rabbit IgG-alkaline phosphatase conjugate with enzymatic activity measured by p-nitrophenylphosphate as substrate. This ELISA detected ACE with a sensitivity of 32 ng/ml cellular ACE. Elevation in cellular ACE catalytic activity as measured by fluorescent assay of detergent extracts from bovine endothelial cells corresponded well with an increase in ACE protein as determined by the ELISA. These results provide direct evidence that increases in catalytic activity of ACE produced in endothelial cells by a variety of agents result from enhancement of the synthesis of ACE protein.
New England Medical Center Hospital, Department of Medicine, Boston, MA 02111.
Y
Y
Dasarathy
J
J
Stevens
J J
JJ
Lanzillo
B L
BL
Fanburg
D015056:1-Methyl-3-isobutylxanthine / Q000494:pharmacology; D000450:Aldosterone / Q000494:pharmacology; D000818:Animals; D003994:Bucladesine / Q000494:pharmacology; D000001:Calcimycin / Q000494:pharmacology; D002417:Cattle; D002478:Cells, Cultured; D005576:Colforsin / Q000494:pharmacology; D003907:Dexamethasone / Q000494:pharmacology; D004730:Endothelium, Vascular / Q000187:drug effects / Q000201:enzymology*; D004797:Enzyme-Linked Immunosorbent Assay; D007700:Kinetics; D007703:Peptidyl-Dipeptidase A / Q000378:metabolism*; D014284:Triiodothyronine / Q000494:pharmacology
D014284:Triiodothyronine; D005576:Colforsin; D000001:Calcimycin; D000450:Aldosterone; D003994:Bucladesine; D003907:Dexamethasone; D007703:Peptidyl-Dipeptidase A; D015056:1-Methyl-3-isobutylxanthine