Identification of vascular renin-binding proteins by chemical cross-linking: inhibition of binding of renin by renin inhibitors.
OBJECTIVE
To investigate the mechanism of tissue uptake of renin.


METHODS
Angiotensin peptide formation in tissues is dependent on kidney-derived renin, leading us to hypothesize that tissues possess a mechanism for uptake of renin from plasma.


METHODS
The binding of [125I]-labelled renin to membranes prepared from various rat tissues was examined. [125I]-labelled renins were cross-linked to membranes with disuccinimidyl suberate and analysed by sodium dodecyl sulphate-polyacrylamide gel electrophoresis followed by autoradiography.


RESULTS
Mesenteric artery membranes bound both [125I]-labelled rat renin and [125I]-labelled mouse submandibular gland renin. Cross-linking experiments showed two bands, one of relative molecular mass approximately 105,000 and the other of approximately 75,000. After taking into account the molecular weight of renin, these bands represent renin-binding proteins of relative molecular mass approximately 70,000 and approximately 40,000, respectively. The highest level of these binding proteins was in the mesenteric artery; lower levels were found in the aorta, lung and renal medulla. Renin-binding proteins were also identified in membranes prepared from cultured rat aortic smooth muscle cells. No binding proteins were identified in the kidney cortex, heart, adrenal capsule, adrenal medulla, peri-aortic brown adipose tissue, uterus or pituitary. Binding of renin to mesenteric artery membranes was prevented by inhibitors of renin enzymatic activity (H-77 and SQ 30697); this effect of H-77 showed a dose-dependence parallel to the inhibition of renin activity by this compound, suggesting that the binding of H-77 to the active site of renin prevents its binding to the membranes.


CONCLUSIONS
These studies provide evidence for a vascular renin-binding mechanism, which may play a role in the generation of angiotensin peptides in vasculature, and may thus be a determinant of blood pressure. Moreover, one of the actions of inhibitors of renin enzymatic activity in vivo may be to prevent the binding of renin to the vasculature.
Campbell
D J
DJ
St Vincent's Institute of Medical Research, Fitzroy, Victoria, Australia.
Valentijn
A J
AJ
D000808:Angiotensinogen / Q000378:metabolism; D000818:Animals; D001808:Blood Vessels / Q000378:metabolism*; D002352:Carrier Proteins / Q000378:metabolism*; D003432:Cross-Linking Reagents; D005260:Female; D008297:Male; D008566:Membranes / Q000378:metabolism; D008638:Mesenteric Arteries / Q000378:metabolism; D007703:Peptidyl-Dipeptidase A / Q000378:metabolism; D010949:Plasma / Q000378:metabolism; D051381:Rats; D017207:Rats, Sprague-Dawley; D012083:Renin / Q000378:metabolism*; D000092502:Renin Inhibitors
D000808:Angiotensinogen; D002352:Carrier Proteins; D003432:Cross-Linking Reagents; D007703:Peptidyl-Dipeptidase A; D012083:Renin; D000092502:Renin Inhibitors