Affinity of angiotensin I-converting enzyme (ACE) inhibitors for N- and C-binding sites of human ACE is different in heart, lung, arteries, and veins.
Angiotensin-converting enzyme (ACE) has two enzymatically active domains: a C-domain in the carboxy terminal region and an N-domain in the amino terminal region. We based the pharmacologic characterization of these sites on the rat testis-lung model. In testis, only a truncate form of ACE is present (C-site), whereas both N- and C-sites are present in lung. In this model, captopril was shown to be N-selective and delaprilat to be C-selective. Ro 31-8472, a cilazapril derivative, and enalaprilat proved to be not site selective. We used these drugs to evaluate the affinity of C and N sites in various human tissues involved in the cardiovascular actions of ACE and used [125I]Ro31-8472 as ligand. The number and affinity of ACE binding sites were 17,680 +/- 2,345 fmol/mg protein (Kd = 0.32 +/- 0.04 nM) in lung, 560 +/- 65 (Kd = 0.36 +/- 0.05 nM) in heart, 237 +/- 51 (Kd = 0.37 +/- 0.06 nM) in coronary artery, 236 +/- 63 (Kd = 0.14 +/- 0.05 nM) in saphenous vein, and 603 +/- 121 (Kd = 0.50 +/- 0.06 nM) in mammary artery. The affinity (pKi) of captopril for the N sites ranged from 9.40 +/- 0.14 (lung) to 8.41 +/- 0.10 (coronary artery). The affinity for the C-site by delaprilat ranged from 9.97 +/- 0.15 (coronary artery) to 9.10 +/- 0.14 (mammary artery). Therefore, the affinity of C- and N-sites of ACE for ACE inhibitor (ACEI) drugs is different according to the organ involved. Because ACE is a glycosylated enzyme and glycosylation is organ dependent, we suggest that organ-specific glycosylation affects the binding characteristics of ACE inhibitors to N- or C-site of human tissular ACE.
Endocrinology Department, L Sacco Hospital (Vialba), Milan, Italy.
M
M
Bevilacqua
T
T
Vago
A
A
Rogolino
F
F
Conci
E
E
Santoli
G
G
Norbiato
D000328:Adult; D000806:Angiotensin-Converting Enzyme Inhibitors / Q000378:metabolism*; D000818:Animals; D001158:Arteries / Q000201:enzymology*; D001665:Binding Sites; D002216:Captopril / Q000378:metabolism; D002462:Cell Membrane / Q000201:enzymology; D003331:Coronary Vessels / Q000201:enzymology; D015773:Enalaprilat / Q000378:metabolism; D005260:Female; D006801:Humans; D066298:In Vitro Techniques; D007457:Iodine Radioisotopes; D008168:Lung / Q000201:enzymology*; D008297:Male; D008323:Mammary Arteries / Q000201:enzymology; D009206:Myocardium / Q000201:enzymology*; D007703:Peptidyl-Dipeptidase A / Q000378:metabolism*; D011724:Pyridazines / Q000378:metabolism; D011869:Radioligand Assay; D051381:Rats; D012501:Saphenous Vein / Q000201:enzymology; D013737:Testis / Q000201:enzymology; D014680:Veins / Q000201:enzymology*
D000806:Angiotensin-Converting Enzyme Inhibitors; D007457:Iodine Radioisotopes; D011724:Pyridazines; C076308:Ro 31-8472; D002216:Captopril; D007703:Peptidyl-Dipeptidase A; D015773:Enalaprilat