Ectopic beta-adrenergic receptor binding sites. possible molecular basis of aberrant catecholamine responsiveness of an adrenocortical tumor adenylate cyclase.
L T Williams
T B Gore
R J Lefkowitz
Abstract
The molecular basis for the aberrant catecholamine responsiveness of the adenylate cyclase of adrenocortical carcinoma 494 was explored. The adenylate cyclase of this corticosteroid-producing, transplanted, adrenal cancer of the rat was stimulated not only by adrenocorticotropic hormone and fluoride, but also by the beta-adrenergic agonist, isoproterenol. The adenylate cyclase of normal adrenal tissue was unresponsive to isoproterenol. Direct binding studies with the specific high affinity B-adrenergic ligand, (-)[3H]dihydroalprenolol, demonstrated the pressure of 0.094 pmol of specific binding sites per milligram of tumor membrane protein. By contrast, normal adrenal membranes contained too few binding sites to accurately measure and study using these techniques. The tumor binding sites had high affinity for (-)[3H] dihydroalprenolol with an equilibrium dissociation constant of 2.1 nM. Adrenergic agonists competed for the binding sites in an order of potency, [(-) isoproterenol greater than (-) epinephrine (-) norepinephrine], paralleling their order of potency as beta-adrenergic agonists. The beta-adrenergic antagonist, (-) propranolol, competed for binding, causing half-mzximal inhibition of specific binding at a concentration of 6 nM. The alpha-adrenergic antagonist, phentolamine, and several catecholamine metabolites and precursors did not effectively compete for the binding sites at high concentrations. Binding was stereospecific, the (+) stereoisomers of beta-adrenergic agonists and antagonists requiring 40- to 300-fold higher concentrations than the corresponding (-) stereoisomers to half maximally inhibit (-) [3H] dihydroalprenolol binding. These results indicate that adrenocortical carcinoma 494 membranes contain beta-adrenergic receptor-binding sites which are not normally present in membranes of adrenal tissue. These ectopic beta-adrenergic receptors presumably confer on the neoplastic tissue the catecholamine sensitivity of its adenylate cyclase.
Full text
PDF
Selected References
These references are in PubMed. This may not be the complete list of references from this article.
- Alexander R. W., Williams L. T., Lefkowitz R. J. Identification of cardiac beta-adrenergic receptors by (minus) [3H]alprenolol binding. Proc Natl Acad Sci U S A. 1975 Apr;72(4):1564–1568. doi: 10.1073/pnas.72.4.1564. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Birnbaumer L., Pohl S. L., Kaumann A. J. Receptors and acceptors: a necessary distinction in hormone binding studies. Adv Cyclic Nucleotide Res. 1974;4(0):239–281. [PubMed] [Google Scholar]
- JOHNSON D. F., SNELL K. C., FRANCOIS D., HEFTMANN E. In vitro metabolism of progesterone-4-C-14 in an adrenocortical carcinoma of the rat. Acta Endocrinol (Copenh) 1961 Jul;37:329–335. doi: 10.1530/acta.0.0370329. [DOI] [PubMed] [Google Scholar]
- LOWRY O. H., ROSEBROUGH N. J., FARR A. L., RANDALL R. J. Protein measurement with the Folin phenol reagent. J Biol Chem. 1951 Nov;193(1):265–275. [PubMed] [Google Scholar]
- Lefkowitz R. J., Roth J., Pricer W., Pastan I. ACTH receptors in the adrenal: specific binding of ACTH-125I and its relation to adenyl cyclase. Proc Natl Acad Sci U S A. 1970 Mar;65(3):745–752. doi: 10.1073/pnas.65.3.745. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Lefkowitz R. J. Stimulation of catecholamine-sensitive adenylate cyclase by 5'-guanylyl-imidodiphosphate. J Biol Chem. 1974 Oct 10;249(19):6119–6124. [PubMed] [Google Scholar]
- Maguire M. E., Wiklund R. A., Anderson H. J., Gilman A. G. Binding of (125I)iodohydroxybenzylpindolol to putative beta-adrenergic receptors of rat glioma cells and other cell clones. J Biol Chem. 1976 Mar 10;251(5):1221–1231. [PubMed] [Google Scholar]
- Mayer S. E. Effects of adrenergic agonists and antagonists on adenylate cyclase activity of dog heart and liver. J Pharmacol Exp Ther. 1972 Apr;181(1):116–125. [PubMed] [Google Scholar]
- Mukherjee C., Caron M. G., Coverstone M., Lefkowitz R. J. Identification of adenylate cyclase-coupled beta-adrenergic receptors in frog erythrocytes with (minus)-[3-H] alprenolol. J Biol Chem. 1975 Jul 10;250(13):4869–4876. [PubMed] [Google Scholar]
- Mukherjee C., Caron M. G., Mullikin D., Lefkowitz R. J. Structure-activity relationships of adenylate cyclase-coupled beta adrenergic receptors: determination by direct binding studies. Mol Pharmacol. 1976 Jan;12(1):16–31. [PubMed] [Google Scholar]
- Ney R. L., Hochella N. J., Grahame-Smith D. G., Dexter R. N., Butcher R. W. Abnormal regulation of adenosine 3',5'-monophosphate and corticosterone formation in an adrenocortical carcinoma. J Clin Invest. 1969 Sep;48(9):1733–1739. doi: 10.1172/JCI106139. [DOI] [PMC free article] [PubMed] [Google Scholar]
- SNELL K. C., STEWART H. L. Variations in histologic pattern and functional effects of a transplantable adrenal cortical carcinoma in intact, hypophysectomized, and newborn rats. J Natl Cancer Inst. 1959 Jun;22(6):1119–1155. [PubMed] [Google Scholar]
- Salomon Y., Londos C., Rodbell M. A highly sensitive adenylate cyclase assay. Anal Biochem. 1974 Apr;58(2):541–548. doi: 10.1016/0003-2697(74)90222-x. [DOI] [PubMed] [Google Scholar]
- Schorr I., Ney R. L. Abnormal hormone responses of an adrenocortical cancer adenyl cyclase. J Clin Invest. 1971 Jun;50(6):1295–1300. doi: 10.1172/JCI106608. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Schorr I., Rathnam P., Saxena B. B., Ney R. L. Multiple specific hormone receptors in the adenylate cyclase of an adrenocortical carcinoma. J Biol Chem. 1971 Sep 25;246(18):5806–5811. [PubMed] [Google Scholar]
- Williams L. T., Jarett L., Lefkowitz R. J. Adipocyte beta-adrenergic receptors. Identification and subcellular localization by (-)-[3H]dihydroalprenolol binding. J Biol Chem. 1976 May 25;251(10):3096–3104. [PubMed] [Google Scholar]
- Williams L. T., Snyderman R., Lefkowitz R. J. Identification of beta-adrenergic receptors in human lymphocytes by (-) (3H) alprenolol binding. J Clin Invest. 1976 Jan;57(1):149–155. doi: 10.1172/JCI108254. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Zatz M., Kebabian J. W., Romero J. A., Lefkowitz R. J., Axelrod J. Pineal beta adrenergic receptor: correlation of binding of 3H-l-alprenolol with stimulation of adenylate cyclase. J Pharmacol Exp Ther. 1976 Mar;196(3):714–722. [PubMed] [Google Scholar]