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J. Biol. Chem., Vol. 265, Issue 1, 195-200, 01, 1990
ME Ullian and SL Linas
In some systems there are spare receptors for hormone action, i.e. only a
fraction of the total number of surface receptors need be occupied by
agonist to elicit maximum cellular responses. The purpose of this study was
to determine the relationship between angiotensin II (AII) surface receptor
number and AII-induced inositol trisphosphate (IP3) formation in
rat-cultured vascular smooth muscle cells. To accomplish this purpose, it
was necessary to develop a method to modulate AII surface receptor number
without activating phospholipase C. Incubation with the putative AII
receptor antagonist Sar1,Leu8-AII (SL) caused reductions in AII surface
receptor number by redistribution of receptors to the cell interior.
However, in contrast to AII, SL did not elicit IP3 responses. By varying
the conditions of incubation with SL, graded (32- 60%) reductions in AII
surface receptor number were achieved. In association with reductions in
surface receptors there were comparable reductions in AII-stimulated IP3
formation. The correlation between receptor number and stimulated IP3
formation was highly linear (r = 0.99, p less than 0.01). To determine if
incubation with AII also caused reductions in stimulated IP3 formation in
proportion to the degree of receptor loss, AII surface receptor number was
decreased by incubation with AII. Despite decreases in AII receptor number
comparable to those achieved with SL, incubation with AII resulted in 2-
fold greater loss of AII-stimulated IP3 formation than did incubation with
SL. We conclude that in vascular smooth muscle cells 1) the AII receptor
antagonist SL stimulates AII receptor trafficking without eliciting IP3
formation, 2) there are no spare AII receptors for phospholipase C-mediated
IP3 formation, and 3) AII desensitization of IP3 formation is mediated by
reductions in surface receptors as well as by post-receptor mechanisms.
Angiotensin II surface receptor coupling to inositol trisphosphate formation in vascular smooth muscle cells
Department of Medicine, University of Colorado Health Sciences Center, Denver 80262.
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