Skip to main content
Log in

Molecular comparison of α2-adrenergic receptors from rat adrenocortical carcinoma and human blood platelet

  • Original Article
  • Published:
Molecular and Cellular Biochemistry Aims and scope Submit manuscript


We have previously described a simple two-step purification technique to isolate α2-adrenergic receptors from the rat adrenocortical carcinoma (Jaiswal, R. K. and Sharma, R. K. (1985) Biochem. Biophys. Res. Commun. 130, 58–64). Utilizing this technique we have now achieved ∼ 77 000-fold purification to apparent homogeneity of α2-adrenergic receptors from human platelets. We have compared the biochemical characteristics of these receptors with those from the rat, which were purified ∼ 40000-fold to homogeneity.

The [125I] receptor proteins from two sources showed: (a) a single radioactive band with a Mr of 64000 as evidenced by one- and two-dimensional sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDSPAGE); and (b) a single symmetrical peak with a pl of 4.2 by isoelectric focusing polyacrylamide gel electrophoresis. Both proteins showed typical α2-adrenergic binding characteristics with specific binding activities of 13.85 nmol/mg and 14.17 nmol/mg protein. These values are close to the theoretical binding activity of 15.6 nmol/mg protein for 1 mol of the ligand binding 1 mol of the receptor protein. These results attest to the purity of the receptors, to its Mr of 64000, and to its acidic nature. However, the peptide maps of the radioiodinated α2-adrenergic receptors from rat adrenocortical carcinoma and human blood platelets reveal some distinct differences which may relate to the differences in the pharmacological specificities between rodent and nonrodent α2-adrenergic receptors.

This is a preview of subscription content, log in via an institution to check access.

Access this article

Price excludes VAT (USA)
Tax calculation will be finalised during checkout.

Instant access to the full article PDF.

Similar content being viewed by others









High Performance Liquid Chromatography


  1. Snell KC, Stewart HL: Variations in histological pattern and functional effects of a transplantable adrenalcortical carcinoma in intact, hypophysectomized and new born rats. J Natl Cancer Inst. 22:1119–1132, 1959

    Google Scholar 

  2. Sharma RK, Hashimoto K: Ultrastructural studies and metabolic regulation of isolated adrenocrotical carcinoma cells of rat. Cancer Res 32:666–671, 1972

    Google Scholar 

  3. Nambi P, Aiyar NV, Sharma RK: Identification and characterization of ectopic α2-adrenergic receptors in adrenocortical carcinoma membranes. J Nutrition, Growth and Cancer 1:77–84, 1983

    Google Scholar 

  4. Nambi P, Aiyar NV, Sharma RK: Solubulization of epinephrine-specific α2-receptors from adrenocortical carcinoma. FEES Letters 140:98–102, 1982

    Google Scholar 

  5. Jaiswal N, Sharma RK: Dual regulation of adenylate cyclase and guanylate cyclase: α2-adrenergic signal transduction in adrenocortical carcinoma cells. Arch Biochem Biophys 249:616–619, 1986

    Google Scholar 

  6. Jaiswal RK, Sharma RK: Purification and biochemical characterization of α2-adrenergic receptor from rat adrenocortical carcinoma. Biochem Biophys Res Commun 130:58–64, 1985

    Google Scholar 

  7. Hunter WH, Greenwood FC: Preparation of iodine-131 labelled human growth hormone of high specific activity. Nature (London) 194:495–496, 1962

    Google Scholar 

  8. O'Farrel PH: High resolution two-dimensional electrophoresis of proteins. J Biol Chem 250:4007–4021, 1975

    Google Scholar 

  9. Cleveland DL, Fischer SG, Kirschner MW, Laemmli UK: Peptide mapping by limited proteolysis in sodium dodecyl sulfate and ananysis by gel electrophoresis. J Biol Chem 252:1102–1106, 1977

    Google Scholar 

  10. Bradford MM: A rapid and sensitive method for the quantitation of microgram quantities of protein utilizing the principle of protein-dye binding. Anal Biochem 72:248–254, 1976

    Article  CAS  PubMed  Google Scholar 

  11. Fried VA, Ando ME, Bel AJ: Protein quantitation at the picomole level: an O-phthaldialdehyde-pre TSK columnderivatization assay. Anal Biochem 146:271–276, 1985

    Google Scholar 

  12. Bidlingmeyer BA, Cohen SA, Tarvin TL: Rapid analysis of amino acids using precolumn derivatization. J Chromatogr 36:93–104, 1984

    Google Scholar 

  13. Regan JW, Barden N, Lefkowitz RJ, Caron MG, DeMarinis RM, Krog AJ, Holden KG, Matthews WD, Hieble JP: Affinity chromatography of human platelets a2-adrenergic receptors. Proc Natl Acad Sci 79:7223–7227, 1982

    Google Scholar 

  14. Kawahara RS, Byington KH, Bylund DB: [3H]pazidoclonidine: a photoaffinity label for the α2-adrenergic receptor. (Abstract) Fed Proc 43:951, 1984

    Google Scholar 

  15. Shreeve MS, Fraser CM, Venter JC: Molecular comparison of α1- and α2-adrenergic receptors suggests that these proteins are structurally related ‘isoreceptors’. Proc Natl Acad Sci 82:4842–4846, 1985

    Google Scholar 

  16. Bylund DB: Heterogeneity of α2-adrenergic receptors. Pharmacol Biochem Behav 22:835–843, 1985

    Google Scholar 

  17. Repaske MG, Nunnari JM, Limbird LE: Purification of the α2-adrenergic receptor from porcine brain using a yohimbine-agarose affinity matrix. J Biol Chem 262:12381–12386, 1987

    Google Scholar 

  18. Regan JW, Nakata H, DeMarinis RM, Caron M, Lefkowitz RJ: Purification and characterization of the human platelets α2-adrenergic receptor. J Biol Chem 261:3894–3900, 1986

    Google Scholar 

  19. Kobilka BK, Matsui H, Kobilka TS, Yang-Feng TL, Francke V, Caron MG, Lefkowitz RJ, Regan JW: Cloning, sequencing and expression of the gene coding for the human platelets α2-adrenergic receptor. Science 238:650–656, 1987

    CAS  PubMed  Google Scholar 

  20. Hoffman BB, Lefkowitz RJ: Radioligand binding studies of adrenergic receptors: new insights into molecular and physiological regulation. Annual Rev Pharmacol Toxicol 20:581–608, 1980

    Google Scholar 

  21. Guicheney P, Garay RP, Levy-Marchal C, Meyer P: Biochemical evidence for presynaptic and postsynaptic α-adrenoreceptors in rat heart membrane: positive homotropic cooperativity of presynaptic binding. Proc Natl Acad Sci USA 75:6285–6289, 1978

    Google Scholar 

  22. Miach PJ, Dansse JP, Meyer P: Direct biochemical demonstration of two types of α-adrenoreceptor in rat brain. Nature (London) 274:492–494, 1979

    Google Scholar 

  23. Pacquery R, Guidicelli Y Heterogeneity and subcellular localization of hamster adipocyte α-adrenergic receptors. FEBS Letter 116:85–90, 1980

    Google Scholar 

  24. Lynch CJ, Steer ML: Evidence for high and low affinity α2-receptors. J Biol Chem 256:3298–3303, 1981

    Google Scholar 

  25. Hoffman BB, Yim S, Tsai BS, Lefkowitz RJ: Preferential uncoupling by manganese of α-adrenergic receptor mediated inhibition of adenylate cyclase in human platelets. Biochem Biophys Res Commun 100:724–731, 1981

    Google Scholar 

  26. Marquis HR, Becker JA, Vigdahel RL: Platelet aggregation III. An epinephrine induced decrease in cyclic AMP synthesis. Biochem Biophys Res Commun 39:783–789, 1970

    Google Scholar 

  27. Paul AK, Marala RB, Jaiswal RK, Sharma RK: Coexistence of guanylate cyclase and atrial natriuretic factor receptor in a 180-kD protein. Science 235:1224–1226, 1987

    Google Scholar 

Download references

Author information

Authors and Affiliations


Rights and permissions

Reprints and permissions

About this article

Cite this article

Jaiswall, R.K., Marshak, D.R. & Sharma, R.K. Molecular comparison of α2-adrenergic receptors from rat adrenocortical carcinoma and human blood platelet. Mol Cell Biochem 86, 41–53 (1989).

Download citation

  • Received:

  • Accepted:

  • Issue Date:

  • DOI:

Key words