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. 1972 Jan;69(1):283–287. doi: 10.1073/pnas.69.1.283

Binding of Thyrotropin-Releasing Hormone to Plasma Membranes of Bovine Anterior Pituitary Gland

Fernand Labrie 1, Nicholas Barden 1, Guy Poirier 1, Andre De Lean 1
PMCID: PMC427592  PMID: 4621548

Abstract

An assay for the binding of [3H]thyrotropin-releasing hormone ([3H]TRH) is described. Plasma membranes isolated from bovine anterior pituitary gland bind about 600 femtomoles of this hormone per mg of protein, as compared to 15 femtomoles per mg of protein in the total adenohypophyseal homogenate (40-fold purification). The equilibrium constant of membrane receptor-[3H]TRH binding at 0°C is 4.3 × 107 L·M-1, or a half-maximal binding of this hormone at 23 nM. The binding is time-dependent; addition of unlabeled hormone induces dissociation of the receptor-[3H]TRH complex with a half-life of 14 min. The binding of TRH is not altered by 10 μM melanocyte-stimulating hormone-release inhibiting hormone, lysine-vasopressin, adrenocorticotropin, growth hormone, prolactin, luteinizing hormone, insulin, glucagon, L-thyroxine, or L-triiodothyronine. K+ and Mg++ increase formation of the receptor-TRH complex at optimal concentrations of 5-25 mM and 0.5-2.5 mM, respectively, with inhibition at higher concentrations. Ca++ inhibits binding of TRH at all concentrations tested.

Keywords: hormone receptor, adenylate cyclase, equilibrium constant, [3H]thyrotropin

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Selected References

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

  1. Boler J., Enzmann F., Folkers K., Bowers C. Y., Schally A. V. The identity of chemical and hormonal properties of the thyrotropin releasing hormone and pyroglutamyl-histidyl-proline amide. Biochem Biophys Res Commun. 1969 Nov 6;37(4):705–710. doi: 10.1016/0006-291x(69)90868-7. [DOI] [PubMed] [Google Scholar]
  2. Bowers C. Y., Lee K. L., Schally A. V. A study on the interaction of the thyrotropin-releasing factor and L-triiodothyronine: effects of puromycin and cycloheximide. Endocrinology. 1968 Jan;82(1):75–82. doi: 10.1210/endo-82-1-75. [DOI] [PubMed] [Google Scholar]
  3. Bowers C. Y., Weil A., Chang J. K., Sievertsson H., Enzmann F., Folkers K. Activity-structure relationships of the thyrotropin releasing hormone. Biochem Biophys Res Commun. 1970 Aug 11;40(3):683–691. doi: 10.1016/0006-291x(70)90958-7. [DOI] [PubMed] [Google Scholar]
  4. Burgus R., Dunn T. F., Desiderio D., Guillemin R. Structure moléculaire du facteur hypothalamique hypophysiotrope TRF d'origine ovine: mise en évidence par spectrométrie de masse de la séquence PCA-His-Pro-NH2. C R Acad Sci Hebd Seances Acad Sci D. 1969 Nov 12;269(19):1870–1873. [PubMed] [Google Scholar]
  5. Burgus R., Dunn T. F., Desiderio D., Ward D. N., Vale W., Guillemin R. Characterization of ovine hypothalamic hypophysiotropic TSH-releasing factor. Nature. 1970 Apr 25;226(5243):321–325. doi: 10.1038/226321a0. [DOI] [PubMed] [Google Scholar]
  6. Burgus R., Guillemin R. Hypothalamic releasing factors. Annu Rev Biochem. 1970;39:499–526. doi: 10.1146/annurev.bi.39.070170.002435. [DOI] [PubMed] [Google Scholar]
  7. Cheever E. V., Lewis U. J. Estimation of the molecular weights of the multiple components of growth hormone and prolactin. Endocrinology. 1969 Sep;85(3):465–473. doi: 10.1210/endo-85-3-465. [DOI] [PubMed] [Google Scholar]
  8. Cuatrecasas P. Insulin--receptor interactions in adipose tissue cells: direct measurement and properties. Proc Natl Acad Sci U S A. 1971 Jun;68(6):1264–1268. doi: 10.1073/pnas.68.6.1264. [DOI] [PMC free article] [PubMed] [Google Scholar]
  9. Fellows R. E., Jr, Rogol A. D. Structural studies on bovine growth hormone. I. Isolation and characterization of cyanogen bromide fragments. J Biol Chem. 1969 Mar 25;244(6):1567–1575. [PubMed] [Google Scholar]
  10. Fleischer N., Donald R. A., Butcher R. W. Involvement of adenosine 3',5'-monophosphate in release of ACTH. Am J Physiol. 1969 Nov;217(5):1287–1291. doi: 10.1152/ajplegacy.1969.217.5.1287. [DOI] [PubMed] [Google Scholar]
  11. Freychet P., Roth J., Neville D. M., Jr Insulin receptors in the liver: specific binding of ( 125 I)insulin to the plasma membrane and its relation to insulin bioactivity. Proc Natl Acad Sci U S A. 1971 Aug;68(8):1833–1837. doi: 10.1073/pnas.68.8.1833. [DOI] [PMC free article] [PubMed] [Google Scholar]
  12. Freychet P., Roth J., Neville D. M., Jr Monoiodoinsulin: demonstration of its biological activity and binding to fat cells and liver membranes. Biochem Biophys Res Commun. 1971 Apr 16;43(2):400–408. doi: 10.1016/0006-291x(71)90767-4. [DOI] [PubMed] [Google Scholar]
  13. House P. D., Weidemann M. J. Characterization of an [125 I]-insulin binding plasma membrane fraction from rat liver. Biochem Biophys Res Commun. 1970 Nov 9;41(3):541–548. doi: 10.1016/0006-291x(70)90046-x. [DOI] [PubMed] [Google Scholar]
  14. 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]
  15. Labrie F., Béraud G., Gauthier M., Lemay A. Actinomycin-insensitive stimulation of protein synthesis in rat anterior pituitary in vitro by dibutyryl adenosine 3',5'-monophosphate. J Biol Chem. 1971 Mar 25;246(6):1902–1908. [PubMed] [Google Scholar]
  16. Labrie F., Pelletier G., Labrie R., Ho-Kim M. A., Delgado A., MacIntosh B., Fortier C. Liaison transcortine-corticostérone et contrôle de l'activité hypophyso-surrénalienne chez le rat. Interactions hypophyse-thyroïde-surrénales-gonades. Ann Endocrinol (Paris) 1968 Jan-Feb;29(1):29–43. [PubMed] [Google Scholar]
  17. Lefkowitz R. J., Roth J., Pastan I. Effects of calcium on ACTH stimulation of the adrenal: separation of hormone binding from adenyl cyclase activation. Nature. 1970 Nov 28;228(5274):864–866. doi: 10.1038/228864a0. [DOI] [PubMed] [Google Scholar]
  18. 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]
  19. Lin S. Y., Goodfriend T. L. Angiotensin receptors. Am J Physiol. 1970 May;218(5):1319–1328. doi: 10.1152/ajplegacy.1970.218.5.1319. [DOI] [PubMed] [Google Scholar]
  20. McCann S. M., Porter J. C. Hypothalamic pituitary stimulating and inhibiting hormones. Physiol Rev. 1969 Apr;49(2):240–284. doi: 10.1152/physrev.1969.49.2.240. [DOI] [PubMed] [Google Scholar]
  21. Mitnick M., Reichlin S. Thyrotropin-releasing hormone: biosynthesis by rat hypothalamic fragments in vitro. Science. 1971 Jun 18;172(3989):1241–1243. doi: 10.1126/science.172.3989.1241. [DOI] [PubMed] [Google Scholar]
  22. NEVILLE D. M., Jr The isolation of a cell membrane fraction from rat liver. J Biophys Biochem Cytol. 1960 Oct;8:413–422. doi: 10.1083/jcb.8.2.413. [DOI] [PMC free article] [PubMed] [Google Scholar]
  23. Robison G. A., Butcher R. W., Sutherland E. W. Cyclic AMP. Annu Rev Biochem. 1968;37:149–174. doi: 10.1146/annurev.bi.37.070168.001053. [DOI] [PubMed] [Google Scholar]
  24. Rodbell M., Krans H. M., Pohl S. L., Birnbaumer L. The glucagon-sensitive adenyl cyclase system in plasma membranes of rat liver. 3. Binding of glucagon: method of assay and specificity. J Biol Chem. 1971 Mar 25;246(6):1861–1871. [PubMed] [Google Scholar]
  25. Rodbell M., Krans H. M., Pohl S. L., Birnbaumer L. The glucagon-sensitive adenyl cyclase system in plasma membranes of rat liver. IV. Effects of guanylnucleotides on binding of 125I-glucagon. J Biol Chem. 1971 Mar 25;246(6):1872–1876. [PubMed] [Google Scholar]
  26. SUTHERLAND E. W., RALL T. W., MENON T. Adenyl cylase. I. Distribution, preparation, and properties. J Biol Chem. 1962 Apr;237:1220–1227. [PubMed] [Google Scholar]
  27. Tomasi V., Koretz S., Ray T. K., Dunnick J., Marinetti G. V. Hormone action at the membrane level. II. The binding of epinephrine and glucagon to the rat liver plasma membrane. Biochim Biophys Acta. 1970 Jul 7;211(1):31–42. doi: 10.1016/0005-2736(70)90120-3. [DOI] [PubMed] [Google Scholar]
  28. Vale W., Burgus R., Guillemin R. Presence of calcium ions as a requisite for the in vitro stimulation of TSH-release by hypothalamic TRF. Experientia. 1967 Oct 15;23(10):853–855. doi: 10.1007/BF02146887. [DOI] [PubMed] [Google Scholar]
  29. Wilber J. F., Peake G. T., Utiger R. D. Thyrotropin release in vitro: stimulation by cyclic 3',5'-Adenosine monophosphate. Endocrinology. 1969 Apr;84(4):758–760. doi: 10.1210/endo-84-4-758. [DOI] [PubMed] [Google Scholar]

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