Degradation of bradykinin by neutral endopeptidase (EC 3.4.24.11) in cultured human endothelial cells.
The presence of neutral endopeptidase 24.11 was demonstrated in human umbilical vein endothelial cells by immunostaining. Enzymatic activity of neutral endopeptidase was determined as 0.167 +/- 0.02 mU/mg protein in the membrane fraction of human umbilical vein endothelial cells, using the fluorogenic peptide substrate, dansyl-D-Ala-Gly-Phe(pNO2)-Gly. No activity was found in the cytosolic fraction of endothelial cells. The role of this peptidase in the degradation of the endogenous vasodilator bradykinin was investigated by incubating human umbilical vein endothelial cell monolayers with bradykinin (10(-8) mol/l). The inhibitor of neutral endopeptidase, phosphoramidon (10(-8) mol/l), decreased the degradation of bradykinin in the supernatant of endothelial cells; the half-life of bradykinin was then increased from 29 +/- 1 to 46 +/- 2 minutes. The angiotensin-converting enzyme inhibitor, lisinopril (10(-8) mol/l), increased the half-life of bradykinin to 244 +/- 20 minutes; the combination of both inhibitors increased the half-life of bradykinin to 381 +/- 51 minutes. Inhibitors of aminopeptidase (amastatin) and carboxypeptidase (2-mercaptomethyl-3-guanidinoethyl-thiopropionic acid) caused no significant effect. The effect of phosphoramidon was small in comparison with that of lisinopril, but was pronounced in combination with lisinopril. Neutral endopeptidase activity is localized in the membranes of human endothelial cells and seems to be involved in the degradation of bradykinin by the vascular endothelium, particularly during angiotensin converting enzyme inhibition.
Department of Cardiology, German Heart Institute Berlin, Germany.
K
K
Graf
M
M
Gräfe
C
C
Bossaller
J
J
Niehus
K D
KD
Schulz
W
W
Auch-Schwelk
E
E
Fleck
D000806:Angiotensin-Converting Enzyme Inhibitors / Q000494:pharmacology; D001920:Bradykinin / Q000378:metabolism*; D002478:Cells, Cultured; D003600:Cytosol / Q000201:enzymology; D004151:Dipeptides / Q000494:pharmacology; D004730:Endothelium, Vascular / Q000201:enzymology*; D006020:Glycopeptides / Q000494:pharmacology; D006207:Half-Life; D006801:Humans; D017706:Lisinopril; D015260:Neprilysin / Q000037:antagonists & inhibitors / Q000378:metabolism*; D013379:Substrate Specificity; D014471:Umbilical Veins
D000806:Angiotensin-Converting Enzyme Inhibitors; D004151:Dipeptides; D006020:Glycopeptides; D017706:Lisinopril; D015260:Neprilysin; D001920:Bradykinin; C008890:phosphoramidon