Calcitonin gene related peptide (CGRP) activates adenylate cyclase of bovine aortic endothelial cells: guanosine 5' triphosphate dependence and partial agonist activity of a tyrosinated analogue.
Human calcitonin gene related peptide (hCGRP) has been shown to release prostacyclin from cultured human endothelial cells and activate adenylate cyclase in endothelial cell membrane preparations. Human endothelial cells are difficult to obtain, so a bovine aortic endothelial cell model has been used to investigate the actions of hCGRP further. Bovine aortic endothelial cells were cultured and membranes prepared from them. In these membrane fractions there was a concentration dependent activation of adenylate cyclase by hCGRP, which required the presence of guanosine 5' triphosphate. Basal activity of adenylate cyclase was 41.5 pmol.min-1.mg-1 protein. The concentration for half maximum activation of adenylate cyclase (Kact) by hCGRP was 842 nM. The maximum increase in adenylate cyclase activity above basal in response to hCGRP was 180 pmol cyclic adenosine monophosphate.min-1.mg-1 protein. A [tyr degree] substituted analogue of hCGRP [( tyr degree]-hCGRP) also activated adenylate cyclase (Kact 2.2 microM) but the maximum stimulation by [tyr degree]-hCGRP was less than that obtained with native peptide. The same analogue partially inhibited the hCGRP dependent activation of adenylate cyclase (Ki = 2 microM). These studies show that hCGRP acts on a receptor on endothelial cells to activate adenylate cyclase and that [tyr degree]-hCGPO is a partial agonist at this receptor. The potent vasodilator properties of hCGRP may be mediated through or modified by its interaction with endothelial cells.
McEwan
J R
JR
Department of Clinical Pharmacology, Royal Postgraduate Medical School, London.
Ritter
J M
JM
MacDermot
J
J
D000262:Adenylyl Cyclases / Q000378:metabolism*; D000818:Animals; D001011:Aorta; D015740:Calcitonin Gene-Related Peptide / Q000031:analogs & derivatives / Q000032:analysis / Q000494:pharmacology*; D002417:Cattle; D004730:Endothelium, Vascular / Q000032:analysis / Q000187:drug effects* / Q000201:enzymology; D004789:Enzyme Activation / Q000187:drug effects*; D006160:Guanosine Triphosphate / Q000494:pharmacology; D011956:Receptors, Cell Surface / Q000502:physiology; D014443:Tyrosine / Q000494:pharmacology
D011956:Receptors, Cell Surface; D014443:Tyrosine; D006160:Guanosine Triphosphate; D000262:Adenylyl Cyclases; D015740:Calcitonin Gene-Related Peptide