Functional modification of the guanine nucleotide regulatory protein after desensitization of turkey erythrocytes by catecholamines

doi:10.1016/0003-9861(83)90198-4

Archives of Biochemistry and Biophysics

Volume 224, Issue 1, 1 July 1983, Pages 142-151

https://doi.org/10.1016/0003-9861(83)90198-4 Get rights and content

Abstract

Densensitization of turkey erythrocytes by exposure to the beta-adrenergic agonist (−)isoproterenol leads to decreased activation of adenylate cyclase by agonist, NaF, and guanyl-5′-yl imido diphosphate, with no reduction in the number of beta-adrenergic receptors. Interactions between the receptor and the guanine nucleotide regulatory protein (N protein) also seem to be impaired. These observations suggest that a component distal to the beta-adrenergic receptor may be a locus of modification. Accordingly we examined the N protein to determine whether it was altered by desensitization. The rate at which (−)isoproterenol stimulated the release of [³H]GDP from the N protein was substantially lower in membranes prepared from desensitized cells, providing further evidence for uncoupling of the receptor and the N protein. The amount of N protein in membranes from control and desensitized cells was compared by labeling the 42,000 M_r component of the N protein with [³²P]NAD⁺ and cholera toxin; no significant difference was found. However, significantly more N protein (p < .001) was solubilized by cholate extraction of desensitized membranes, suggesting an altered association of the N protein with the membrane after desensitization. The functional activity of the N protein was measured by reconstitution of cholate extracts of turkey erythrocyte membranes into S49 lymphoma cyc⁻ membranes. Reconstitution of (−)isoproterenol stimulation of adenylate cyclase activity was reduced significantly (p < .05) after desensitization. These observations suggest that desensitization of the turkey erythrocyte by (−)isoproterenol results in functional modifications of the guanine nucleotide regulatory protein, leading to impaired interactions with the beta-adrenergic receptor and reduced activation of adenylate cyclase.

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Functional modification of the guanine nucleotide regulatory protein after desensitization of turkey erythrocytes by catecholamines

Abstract

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