Assembly and activation of HK-PK complex on endothelial cells results in bradykinin liberation and NO formation.
Prekallikrein (PK) activation on human umbilical endothelial cells (HUVEC) presumably leads to bradykinin liberation. On HUVEC, PK activation requires the presence of cell-bound high-molecular-weight kininogen (HK) and Zn(2+). We examined the Zn(2+) requirement for HK binding to and the consequences of PK activation on endothelial cells. Optimal HK binding (14 pmol/10(6) HUVEC) is seen with no added Zn(2+) in HEPES-Tyrode buffer containing gelatin versus 16--32 microM added Zn(2+) in the same buffer containing bovine serum albumin. The affinity and number of HK binding sites on HUVEC are a dissociation constant of 9.6 +/- 1.8 nM and a maximal binding of 1.08 +/- 0.26 x 10(7) sites/cell (means +/- SD). PK is activated to kallikrein by an antipain-sensitive mechanism in the presence of HK and Zn(2+) on HUVEC, human microvascular endothelial cells, umbilical artery smooth muscle cells, and bovine pulmonary artery endothelial cells. Simultaneous with kallikrein formation, bradykinin (5.0 or 10.3 pmol/10(6) HUVEC in the absence or presence of lisinopril, respectively) is liberated from cell-bound HK. Liberated bradykinin stimulates the endothelial cell bradykinin B2 receptor to form nitric oxide. Assembly and activation of PK on endothelial cells modulates their physiological activities.
Division of Hematology and Oncology, Department of Internal Medicine, University of Michigan, Ann Arbor, Michigan 48109-5669, USA.
Y
Y
Zhao
Q
Q
Qiu
F
F
Mahdi
Z
Z
Shariat-Madar
R
R
Røjkjaer
A H
AH
Schmaier
D000818:Animals; D000976:Antipain / Q000494:pharmacology; D001665:Binding Sites; D019921:Biotinylation; D001920:Bradykinin / Q000378:metabolism*; D002417:Cattle; D002478:Cells, Cultured; D004730:Endothelium, Vascular / Q000378:metabolism*; D004789:Enzyme Activation; D006801:Humans; D007610:Kallikreins / Q000378:metabolism*; D007700:Kinetics; D019679:Kininogen, High-Molecular-Weight / Q000378:metabolism*; D017706:Lisinopril / Q000494:pharmacology; D008833:Microcirculation; D019331:NG-Nitroarginine Methyl Ester / Q000494:pharmacology; D009569:Nitric Oxide / Q000378:metabolism*; D011288:Prekallikrein / Q000378:metabolism*; D011651:Pulmonary Artery; D018002:Receptors, Bradykinin / Q000502:physiology; D012710:Serum Albumin, Bovine; D014469:Umbilical Arteries; D014471:Umbilical Veins; D015032:Zinc / Q000494:pharmacology; D019323:omega-N-Methylarginine / Q000494:pharmacology
D019679:Kininogen, High-Molecular-Weight; D018002:Receptors, Bradykinin; D012710:Serum Albumin, Bovine; D019323:omega-N-Methylarginine; D009569:Nitric Oxide; D000976:Antipain; D011288:Prekallikrein; D017706:Lisinopril; D007610:Kallikreins; D015032:Zinc; D001920:Bradykinin; D019331:NG-Nitroarginine Methyl Ester