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. 1988 Feb 15;250(1):215–219. doi: 10.1042/bj2500215

A comparison of lactogenic receptors from rat liver and Nb2 rat lymphoma cells by using cross-linking techniques.

C F Webb 1, M Wallis 1
PMCID: PMC1148835  PMID: 3355513

Abstract

Lactogenic receptors were analysed with the use of the cross-linking agent disuccinimidyl suberate to attach covalently 125I-labelled ovine prolactin or human growth hormone to binding sites from (1) liver from pregnant rats and (2) the rat-derived Nb2 lymphoma cell line. Analysis by SDS/polyacrylamide-gel electrophoresis of the proteins cross-linked to labelled hormone in rat liver indicated a major specifically-labelled complex with an Mr of 68,000-72,000, when run under reducing or non-reducing conditions. With Nb2 cells a major specifically-labelled complex with an Mr of 97,000-110,000 was identified, but only when electrophoresis was run using reducing conditions. Assuming one hormone molecule (Mr 22,000-24,000) per hormone-receptor complex, then the receptor proteins have an Mr of 44,000-50,000 for rat liver and 73,000-88,000 for the Nb2 cells. For both cell types the receptors were of lactogenic specificity; lactogenic hormones competed for binding whereas somatogenic hormones did not. These studies suggest that the lactogenic receptors in rat liver membranes and Nb2 cells differ in two respects. Firstly, the Mr of the labelled receptor protein in Nb2 cells is greater than that of the corresponding receptor protein in rat liver membranes; secondly, the Nb2 cell receptor appears to exist as a disulphide-linked oligomer whereas the receptor in rat liver membranes does not.

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Selected References

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

  1. Amit T., Barkey R. J., Gavish M., Youdim M. B. Induction of prolactin (PRL) receptors by PRL in the rat lung and liver. Demonstration and characterization of a soluble receptor. Endocrinology. 1984 Feb;114(2):545–552. doi: 10.1210/endo-114-2-545. [DOI] [PubMed] [Google Scholar]
  2. Ashkenazi A., Cohen R., Gertler A. Characterization of lactogen receptors in lactogenic hormone-dependent and independent NB2 lymphoma cell lines. FEBS Lett. 1987 Jan 1;210(1):51–55. doi: 10.1016/0014-5793(87)81296-6. [DOI] [PubMed] [Google Scholar]
  3. Bohnet H. G., Gómez F., Friesen H. G. Prolactin and estrogen binding sites in the mammary gland of the lactating and non-lactating rat. Endocrinology. 1977 Oct;101(4):1111–1121. doi: 10.1210/endo-101-4-1111. [DOI] [PubMed] [Google Scholar]
  4. Borst D. W., Sayare M., Posner B. I. Characterization of the prolactin receptor in cell fractions from rat liver. Mol Cell Endocrinol. 1985 Feb;39(2):125–130. doi: 10.1016/0303-7207(85)90129-7. [DOI] [PubMed] [Google Scholar]
  5. Cadman H. F., Wallis M. An investigation of sites that bind human somatotropin (growth hormone) in the liver of the pregnant rabbit. Biochem J. 1981 Sep 15;198(3):605–614. doi: 10.1042/bj1980605. [DOI] [PMC free article] [PubMed] [Google Scholar]
  6. Calvo J. C., Finocchiaro L., Lüthy I., Charreau E. H., Calandra R. S., Engström B., Hansson V. Specific prolactin binding in the rat adrenal gland: its characterization and hormonal regulation. J Endocrinol. 1981 May;89(2):317–325. doi: 10.1677/joe.0.0890317. [DOI] [PubMed] [Google Scholar]
  7. Fraker P. J., Speck J. C., Jr Protein and cell membrane iodinations with a sparingly soluble chloroamide, 1,3,4,6-tetrachloro-3a,6a-diphrenylglycoluril. Biochem Biophys Res Commun. 1978 Feb 28;80(4):849–857. doi: 10.1016/0006-291x(78)91322-0. [DOI] [PubMed] [Google Scholar]
  8. Gout P. W., Beer C. T., Noble R. L. Prolactin-stimulated growth of cell cultures established from malignant Nb rat lymphomas. Cancer Res. 1980 Jul;40(7):2433–2436. [PubMed] [Google Scholar]
  9. Hughes J. P., Simpson J. S., Friesen H. G. Analysis of growth hormone and lactogenic binding sites cross-linked to iodinated human growth hormone. Endocrinology. 1983 Jun;112(6):1980–1985. doi: 10.1210/endo-112-6-1980. [DOI] [PubMed] [Google Scholar]
  10. Kelly P. A., Posner B. I., Tsushima T., Friesen H. G. Studies of insulin, growth hormone and prolactin binding: ontogenesis, effects of sex and pregnancy. Endocrinology. 1974 Aug;95(2):532–539. doi: 10.1210/endo-95-2-532. [DOI] [PubMed] [Google Scholar]
  11. Mitani M., Dufau M. L. Purification and characterization of prolactin receptors from rat ovary. J Biol Chem. 1986 Jan 25;261(3):1309–1315. [PubMed] [Google Scholar]
  12. Mountjoy K., Cowden E. A., Dobbie J. W., Ratcliffe J. G. Prolactin receptors in the rat kidney. J Endocrinol. 1980 Oct;87(1):47–54. doi: 10.1677/joe.0.0870047. [DOI] [PubMed] [Google Scholar]
  13. Posner B. I., Kelly P. A., Shiu R. P., Friesen H. G. Studies of insulin, growth hormone and prolactin binding: tissue distribution, species variation and characterization. Endocrinology. 1974 Aug;95(2):521–531. doi: 10.1210/endo-95-2-521. [DOI] [PubMed] [Google Scholar]
  14. Postel-Vinay M. C. Binding of human growth hormone to rat liver membranes: lactogenic and somatotropic sties. FEBS Lett. 1976 Oct 15;69(1):137–140. doi: 10.1016/0014-5793(76)80670-9. [DOI] [PubMed] [Google Scholar]
  15. Shiu R. P., Elsholtz H. P., Tanaka T., Friesen H. G., Gout P. W., Beer C. T., Noble R. L. Receptor-mediated mitogenic action of prolactin in a rat lymphoma cell line. Endocrinology. 1983 Jul;113(1):159–165. doi: 10.1210/endo-113-1-159. [DOI] [PubMed] [Google Scholar]
  16. Tanaka T., Shiu R. P., Gout P. W., Beer C. T., Noble R. L., Friesen H. G. A new sensitive and specific bioassay for lactogenic hormones: measurement of prolactin and growth hormone in human serum. J Clin Endocrinol Metab. 1980 Nov;51(5):1058–1063. doi: 10.1210/jcem-51-5-1058. [DOI] [PubMed] [Google Scholar]
  17. Tsushima T., Friesen H. G. Radioreceptor assay for growth hormone. J Clin Endocrinol Metab. 1973 Aug;37(2):334–337. doi: 10.1210/jcem-37-2-334. [DOI] [PubMed] [Google Scholar]
  18. Webb C. F., Cadman H. F., Wallis M. The specificity of binding of growth hormone and prolactin to purified plasma membranes from pregnant-rabbit liver. Biochem J. 1986 Jun 15;236(3):657–663. doi: 10.1042/bj2360657. [DOI] [PMC free article] [PubMed] [Google Scholar]
  19. Yamada K., Donner D. B. Structures of the somatotropin receptor and prolactin receptor on rat hepatocytes characterized by affinity labelling. Biochem J. 1984 Jun 1;220(2):361–369. doi: 10.1042/bj2200361. [DOI] [PMC free article] [PubMed] [Google Scholar]

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