Modulation of angiotensin II binding affinity by allosteric interaction of polyvinyl sulfate with an intracellular domain of the DuP-753-sensitive angiotensin II receptor of bovine adrenal glomerulosa.
Angiotensin II (AII) is an important regulator of aldosterone secretion by adrenal glomerulosa cells. All interacts with a specific receptor coupled to a guanine nucleotide-binding protein that controls the activity of phospholipase C. Recently, novel All nonpeptide antagonists (DuP-753 and PD-123319) have been shown to discriminate between two subclasses of All receptors in many different tissues. Our studies confirmed that 125I-All specifically labeled two classes of binding sites for All in a membrane preparation of bovine adrenal glomerulosa cells. The first class (DuP-753 sensitive) represented approximately 85% of the total binding sites for All and possessed a high affinity (IC50 of 92.9 +/- 19.5 nM) for DuP-753. PD-123319 did not have any effect on 125I-All binding to this site. The second class of binding sites was more sensitive to PD-123319, with an IC50 of 6.9 +/- 3.7 nM, and had a much lower affinity for DuP-753 (IC50 around 10 microM). The two classes of receptors had different affinities for All. All showed an affinity around 2 nM for All type 1 receptor (AT1)(DuP-753 sensitive) and a higher affinity, around 0.3 nM, for All type 2 receptor (AT2) (PD-123319 sensitive). All-induced steroidogenesis was completely abolished in the presence of 3 microM DuP-753, indicating that this activity was mediated through a DuP-753-sensitive receptor. We also found that polyvinyl sulfate (PVS), a polyanion, could partly inhibit the binding of 125I-All to bovine adrenal glomerulosa cell membranes, with half-maximal efficiency at 17.3 +/- 8.2 nM. The inhibitory effect of PVS was selective for AT1. The inhibitory effect of PVS was due to a change in the affinity state of the receptor. Unexpectedly, PVS had no effect on All-induced steroidogenesis or on All binding to intact bovine adrenal glomerulosa cells. However, the inhibitory effect of PVS on All binding was recovered after permeabilization of cells. Direct interaction of polyanions with AT1 was suggested by the capacity of solubilized photoaffinity-labeled 125I-AT1 to adsorb to heparin-agarose gels. The adsorption of 125I-AT1 to heparin-agarose was inhibited by prior incubation of solubilized receptor with heparin or PVS. These results suggest that All-induced steroidogenesis is mediated by a DuP-753-sensitive receptor and that PVS decreases the affinity of this receptor by interacting with an intracellular domain (possibly the positively charged domain responsible for coupling with guanine nucleotide-binding proteins).
Boulay
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G
Department of Pharmacology, Faculty of Medicine, University of Sherbrooke, Quebec, Canada.
Servant
G
G
Luong
T T
TT
Escher
E
E
Guillemette
G
G
D000494:Allosteric Regulation; D000804:Angiotensin II / Q000037:antagonists & inhibitors / Q000378:metabolism*; D057911:Angiotensin Receptor Antagonists; D000818:Animals; D001713:Biphenyl Compounds / Q000378:metabolism / Q000494:pharmacology*; D002417:Cattle; D002478:Cells, Cultured; D007093:Imidazoles / Q000378:metabolism / Q000494:pharmacology*; D019808:Losartan; D008861:Microsomes / Q000187:drug effects / Q000378:metabolism; D011145:Polyvinyls / Q000378:metabolism / Q000494:pharmacology*; D011725:Pyridines / Q000494:pharmacology; D011945:Receptors, Angiotensin / Q000378:metabolism*; D015398:Signal Transduction / Q000187:drug effects; D013379:Substrate Specificity; D013777:Tetrazoles / Q000378:metabolism / Q000494:pharmacology*; D015384:Zona Glomerulosa / Q000166:cytology / Q000187:drug effects / Q000378:metabolism*
D057911:Angiotensin Receptor Antagonists; D001713:Biphenyl Compounds; D007093:Imidazoles; D011145:Polyvinyls; D011725:Pyridines; D011945:Receptors, Angiotensin; D013777:Tetrazoles; D000804:Angiotensin II; C073402:PD 123319; C019050:polyvinyl sulfate; D019808:Losartan