Skip to main content
NCBI home page
Proceedings of the National Academy of Sciences of the United States of America logoLink to Proceedings of the National Academy of Sciences of the United States of America
. 1985 Jul;82(14):4842–4846. doi: 10.1073/pnas.82.14.4842

Molecular comparison of alpha 1- and alpha 2-adrenergic receptors suggests that these proteins are structurally related "isoreceptors".

S M Shreeve, C M Fraser, J C Venter
PMCID: PMC391001  PMID: 2862630

Abstract

The structures of human platelet alpha 2-adrenergic receptors and rat liver alpha 1-adrenergic receptors were compared by utilizing isoelectric focusing, NaDodSO4/PAGE, and monoclonal antibody crossreactivity. Digitonin-solubilized alpha 1- and alpha 2-adrenergic receptors have an identical isoelectric point of 4.6. Under reducing conditions in NaDodSO4/polyacrylamide gels, the alpha 1-adrenergic receptor has an apparent molecular mass of 85 kDa. Similarly, the alpha 2-adrenergic receptor, which had been affinity-labeled with [3H]phenoxybenzamine and partially purified by isoelectric focusing or photoaffinity-labeled with p-[3,5-3H]azidoclonidine, was also found to have an apparent molecular mass of 85 kDa. One hybridoma, developed from a fusion between SP2/O myeloma cells and splenic lymphocytes from BALB/c mice immunized with human platelet alpha 2-adrenergic receptors, secreted a monoclonal antibody (alpha 2-116p) against the ligand binding site of alpha 2-adrenergic but not alpha 1-adrenergic receptors. In contrast, three monoclonal antibodies raised against the alpha 1-receptor polypeptide backbone but not the ligand binding site were found to specifically immunoprecipitate human platelet alpha 2-adrenergic receptors. These data suggest that the alpha 1- and alpha 2-adrenergic receptors are "isoreceptors," sharing immunogenic and, by implication, structural determinants that most likely evolved as a result of gene duplication.

Full text

PDF
4842

Selected References

These references are in PubMed. This may not be the complete list of references from this article.

  1. Dubocovich M. L., Langer S. Z. Negative feed-back regulation of noradrenaline release by nerve stimulation in the perfused cat's spleen: differences in potency of phenoxybenzamine in blocking the pre- and post-synaptic adrenergic receptors. J Physiol. 1974 Mar;237(3):505–519. doi: 10.1113/jphysiol.1974.sp010495. [DOI] [PMC free article] [PubMed] [Google Scholar]
  2. FERGUSON K. A. STARCH-GEL ELECTROPHORESIS--APPLICATION TO THE CLASSIFICATION OF PITUITARY PROTEINS AND POLYPEPTIDES. Metabolism. 1964 Oct;13:SUPPL–SUPPL1002. doi: 10.1016/s0026-0495(64)80018-4. [DOI] [PubMed] [Google Scholar]
  3. Fraser C. M., Venter J. C. Monoclonal antibodies to beta-adrenergic receptors: use in purification and molecular characterization of beta receptors. Proc Natl Acad Sci U S A. 1980 Dec;77(12):7034–7038. doi: 10.1073/pnas.77.12.7034. [DOI] [PMC free article] [PubMed] [Google Scholar]
  4. Hoffman B. B., Dukes D. F., Lefkowitz R. J. Alpha-adrenergic receptors in liver membranes: delineation with subtype selective radioligands. Life Sci. 1981 Jan 19;28(3):265–272. doi: 10.1016/0024-3205(81)90732-3. [DOI] [PubMed] [Google Scholar]
  5. Kunos G., Kan W. H., Greguski R., Venter J. C. Selective affinity labeling and molecular characterization of hepatic alpha 1-adrenergic receptors with [3H]phenoxybenzamine. J Biol Chem. 1983 Jan 10;258(1):326–332. [PubMed] [Google Scholar]
  6. Lands A. M., Arnold A., McAuliff J. P., Luduena F. P., Brown T. G., Jr Differentiation of receptor systems activated by sympathomimetic amines. Nature. 1967 May 6;214(5088):597–598. doi: 10.1038/214597a0. [DOI] [PubMed] [Google Scholar]
  7. Langer S. Z. Presynaptic regulation of catecholamine release. Biochem Pharmacol. 1974 Jul 1;23(13):1793–1800. doi: 10.1016/0006-2952(74)90187-7. [DOI] [PubMed] [Google Scholar]
  8. Meier K. E., Sternfeld D. R., Insel P. A. Alpha 1- and beta 2-adrenergic receptors co-expressed on cloned MDCK cells are distinct glycoproteins. Biochem Biophys Res Commun. 1984 Jan 13;118(1):73–81. doi: 10.1016/0006-291x(84)91069-6. [DOI] [PubMed] [Google Scholar]
  9. Motulsky H. J., Shattil S. J., Insel P. A. Characterization of alpha 2-adrenergic receptors on human platelets using [3H]yohimbine. Biochem Biophys Res Commun. 1980 Dec 31;97(4):1562–1570. doi: 10.1016/s0006-291x(80)80044-1. [DOI] [PubMed] [Google Scholar]
  10. Newman K. D., Williams L. T., Bishopric N. H., Lefkowitz R. J. Identification of alpha-adrenergic receptors in human platelets by [3H]dihydroergocryptine binding. J Clin Invest. 1978 Feb;61(2):395–402. doi: 10.1172/JCI108950. [DOI] [PMC free article] [PubMed] [Google Scholar]
  11. Regan J. W., DeMarinis R. M., Caron M. G., Lefkowitz R. J. Identification of the subunit-binding site of alpha 2-adrenergic receptors using [3H]phenoxybenzamine. J Biol Chem. 1984 Jun 25;259(12):7864–7869. [PubMed] [Google Scholar]
  12. Shreeve S. M., Roeske W. R., Venter J. C. Partial functional reconstitution of the cardiac muscarinic cholinergic receptor. J Biol Chem. 1984 Oct 25;259(20):12398–12402. [PubMed] [Google Scholar]
  13. Smith S. K., Limbird L. E. Evidence that human platelet alpha-adrenergic receptors coupled to inhibition of adenylate cyclase are not associated with the subunit of adenylate cyclase ADP-ribosylated by cholera toxin. J Biol Chem. 1982 Sep 10;257(17):10471–10478. [PubMed] [Google Scholar]
  14. Smith S. K., Limbird L. E. Solubilization of human platelet alpha-adrenergic receptors: evidence that agonist occupancy of the receptor stabilizes receptor--effector interactions. Proc Natl Acad Sci U S A. 1981 Jul;78(7):4026–4030. doi: 10.1073/pnas.78.7.4026. [DOI] [PMC free article] [PubMed] [Google Scholar]
  15. Starke K. Alpha-adrenoceptor subclassification. Rev Physiol Biochem Pharmacol. 1981;88:199–236. [PubMed] [Google Scholar]
  16. Steer M. L., Khorana J., Galgoci B. Quantitation and characterization of human platelet alpha-adrenergic receptors using [3H]phentolamine. Mol Pharmacol. 1979 Nov;16(3):719–728. [PubMed] [Google Scholar]
  17. Venter J. C., Eddy B., Hall L. M., Fraser C. M. Monoclonal antibodies detect the conservation of muscarinic cholinergic receptor structure from Drosophila to human brain and detect possible structural homology with alpha 1-adrenergic receptors. Proc Natl Acad Sci U S A. 1984 Jan;81(1):272–276. doi: 10.1073/pnas.81.1.272. [DOI] [PMC free article] [PubMed] [Google Scholar]
  18. Venter J. C., Horne P., Eddy B., Greguski R., Fraser C. M. Alpha 1-adrenergic receptor structure. Mol Pharmacol. 1984 Sep;26(2):196–205. [PubMed] [Google Scholar]
  19. Venter J. C., Schaber J. S., U'Prichard D. C., Fraser C. M. Molecular size of the human platelet alpha 2-adrenergic receptor as determined by radiation inactivation. Biochem Biophys Res Commun. 1983 Nov 15;116(3):1070–1075. doi: 10.1016/s0006-291x(83)80251-4. [DOI] [PubMed] [Google Scholar]

Articles from Proceedings of the National Academy of Sciences of the United States of America are provided here courtesy of National Academy of Sciences

RESOURCES