Solubilization and reconstitution of vasopressin V1 receptors of rat liver

N Aiyar, P Nambi, F Stassen and ST Crooke

The observation that vasopressin receptors loose their binding capacity on solubilization suggests that lipid-milieu may be a requirement for the binding of vasopressin to these receptors. To preserve the ligand binding capacity, we have developed reconstitution procedures to study solubilized rat liver vasopressin receptors. We report here that vasopressin V1 receptors were efficiently solubilized from rat liver membranes with egg lysolecithin and quantitatively reconstituted into phospholipid vesicles resulting in a 4-fold purification. Binding of [3H]arginine vasopressin to the reconstituted receptors was saturable and of high affinity (Kd = 0.6 +/- 0.1 nM). In competition binding experiments the solubilized, reconstituted receptors exhibited the same pharmacological profile as was observed with vasopressin V1 receptors in membranes.

Volume 32, Issue 1, pp. 34-36, 07/01/1987
Copyright © 1987 by American Society for Pharmacology and Experimental Therapeutics