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(Arteriosclerosis, Thrombosis, and Vascular Biology. 2001;21:911.)
© 2001 American Heart Association, Inc.

Vascular Biology

Prorenin Accumulation and Activation in Human Endothelial Cells

Importance of Mannose 6-Phosphate Receptors

Mark M. E. D. van den Eijnden; Jasper J. Saris; René J. A. de Bruin; Elly de Wit; Wim Sluiter; Timothy L. Reudelhuber; Maarten A. D. H. Schalekamp; Frans H. M. Derkx; A. H. Jan Danser

From the Cardiovascular Research Institute COEUR, Departments of Pharmacology (M.M.E.D.v.d.E., J.J.S., A.H.J.D.), Internal Medicine (M.M.E.D.v.d.E., J.J.S., R.J.A.d.B., M.A.D.H.S., F.H.M.D.), and Biochemistry (E.d.W., W.S.), Erasmus University Rotterdam, Rotterdam, the Netherlands, and the Laboratory of Molecular Biochemistry of Hypertension (T.L.R.), Clinical Research Institute of Montréal, Montréal, Canada.

Correspondence to Dr A.H.J. Danser, Department of Pharmacology, Room EE1418b, Erasmus University Rotterdam, Dr. Molewaterplein 50, 3015 GE Rotterdam, Netherlands. E-mail danser{at}farma.fgg.eur.nl

Abstract—ACE inhibitors improve endothelial dysfunction, possibly by blocking endothelial angiotensin production. Prorenin, through its binding and activation by endothelial mannose 6-phosphate (M6P) receptors, may contribute to this production. Here, we investigated this possibility as well as prorenin activation kinetics, the nature of the prorenin-activating enzyme, and M6P receptor–independent prorenin binding. Human umbilical vein endothelial cells (HUVECs) were incubated with wild-type prorenin, K/A-2 prorenin (in which Lys42 is mutated to Ala, thereby preventing cleavage by known proteases), M6P-free prorenin, and nonglycosylated prorenin, with or without M6P, protease inhibitors, or angiotensinogen. HUVECs bound only M6P-containing prorenin (K_d 0.9±0.1 nmol/L, maximum number of binding sites [B_max] 1010±50 receptors/cell). At 37°C, because of M6P receptor recycling, the amount of prorenin internalized via M6P receptors was >25 times B_max. Inside the cells, wild-type and K/A-2 prorenin were proteolytically activated to renin. Renin was subsequently degraded. Protease inhibitors interfered with the latter but not with prorenin activation, thereby indicating that the activating enzyme is different from any of the known prorenin-activating enzymes. Incubation with angiotensinogen did not lead to endothelial angiotensin generation, inasmuch as HUVECs were unable to internalize angiotensinogen. Most likely, therefore, in the absence of angiotensinogen synthesis or endocytosis, M6P receptor–mediated prorenin internalization by endothelial cells represents prorenin clearance.

Key Words: human umbilical vein endothelial cells • mannose 6-phosphate receptors • prorenin • renin • angiotensin

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