Skip to main content
PMC 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
. 1988 Jul;85(13):4837–4841. doi: 10.1073/pnas.85.13.4837

Purification and characterization of the human interferon-gamma receptor from placenta.

J Calderon 1, K C Sheehan 1, C Chance 1, M L Thomas 1, R D Schreiber 1
PMCID: PMC280531  PMID: 2455298

Abstract

Purification of the human interferon-gamma (IFN-gamma) receptor was facilitated by identification of human placenta as a large-scale receptor source. When analyzed in radioligand binding experiments, intact placental membranes and detergent-solubilized membrane proteins expressed 1.3 and 5.9 X 10(12) receptors per mg of protein, respectively, values that were 13-163 times greater than that observed for U937 membranes. Two protocols were followed to purify the IFN-gamma receptor from octyl glucoside-solubilized membranes: (i) sequential affinity chromatography over wheat germ agglutinin- and IFN-gamma-Sepharose and (ii) affinity chromatography over columns containing receptor-specific monoclonal antibody and wheat germ agglutinin. Both procedures resulted in fully active preparations that were 70-90% pure. Purified receptor migrated as a single molecular species of 90 kDa either when analyzed on silver-stained NaDodSO4/polyacrylamide gels or when subjected to electrophoretic transfer blot analysis using a labeled IFN-gamma receptor-specific monoclonal antibody. The identity of the 90-kDa component as the receptor was confirmed by demonstrating its ability to specifically bind 125I-labeled IFN-gamma following NaDodSO4/PAGE and transfer to nitrocellulose. Certain receptor preparations converted into a 55-kDa fragment either during purification or upon storage at 4 degrees C. On the basis of N-Glycanase digestion studies, the IFN-gamma receptor appeared to contain 17 kDa of N-linked carbohydrate. The ligand binding site, the epitope for the receptor-specific monoclonal antibody, and all of the N-linked carbohydrate could be localized to the 55-kDa domain of the molecule.

Full text

PDF
4840

Images in this article

4839Image
on p.4839
4839Image
on p.4839
4839Image
on p.4839
4839Image
on p.4839
4839Image
on p.4839
4839Image
on p.4839
4839Image
on p.4839
4840Image
on p.4840
4840Image
on p.4840

Selected References

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

  1. Aguet M., Merlin G. Purification of human gamma interferon receptors by sequential affinity chromatography on immobilized monoclonal antireceptor antibodies and human gamma interferon. J Exp Med. 1987 Apr 1;165(4):988–999. doi: 10.1084/jem.165.4.988. [DOI] [PMC free article] [PubMed] [Google Scholar]
  2. Allavena P., Scala G., Djeu J. Y., Procopio A. D., Oppenheim J. J., Herberman R. B., Ortaldo J. R. Production of multiple cytokines by clones of human large granular lymphocytes. Cancer Immunol Immunother. 1985;19(2):121–126. doi: 10.1007/BF00199719. [DOI] [PMC free article] [PubMed] [Google Scholar]
  3. Anderson P., Yip Y. K., Vilcek J. Specific binding of 125I-human interferon-gamma to high affinity receptors on human fibroblasts. J Biol Chem. 1982 Oct 10;257(19):11301–11304. [PubMed] [Google Scholar]
  4. Arenzana-Seisdedos F., Virelizier J. L., Fiers W. Interferons as macrophage-activating factors. III. Preferential effects of interferon-gamma on the interleukin 1 secretory potential of fresh or aged human monocytes. J Immunol. 1985 Apr;134(4):2444–2448. [PubMed] [Google Scholar]
  5. Basham T. Y., Merigan T. C. Recombinant interferon-gamma increases HLA-DR synthesis and expression. J Immunol. 1983 Apr;130(4):1492–1494. [PubMed] [Google Scholar]
  6. Beutler B., Tkacenko V., Milsark I., Krochin N., Cerami A. Effect of gamma interferon on cachectin expression by mononuclear phagocytes. Reversal of the lpsd (endotoxin resistance) phenotype. J Exp Med. 1986 Nov 1;164(5):1791–1796. doi: 10.1084/jem.164.5.1791. [DOI] [PMC free article] [PubMed] [Google Scholar]
  7. Celada A., Allen R., Esparza I., Gray P. W., Schreiber R. D. Demonstration and partial characterization of the interferon-gamma receptor on human mononuclear phagocytes. J Clin Invest. 1985 Dec;76(6):2196–2205. doi: 10.1172/JCI112228. [DOI] [PMC free article] [PubMed] [Google Scholar]
  8. Celada A., Gray P. W., Rinderknecht E., Schreiber R. D. Evidence for a gamma-interferon receptor that regulates macrophage tumoricidal activity. J Exp Med. 1984 Jul 1;160(1):55–74. doi: 10.1084/jem.160.1.55. [DOI] [PMC free article] [PubMed] [Google Scholar]
  9. Coffman R. L., Carty J. A T cell activity that enhances polyclonal IgE production and its inhibition by interferon-gamma. J Immunol. 1986 Feb 1;136(3):949–954. [PubMed] [Google Scholar]
  10. Collart M. A., Belin D., Vassalli J. D., de Kossodo S., Vassalli P. Gamma interferon enhances macrophage transcription of the tumor necrosis factor/cachectin, interleukin 1, and urokinase genes, which are controlled by short-lived repressors. J Exp Med. 1986 Dec 1;164(6):2113–2118. doi: 10.1084/jem.164.6.2113. [DOI] [PMC free article] [PubMed] [Google Scholar]
  11. Cuatrecasas P. Isolation of the insulin receptor of liver and fat-cell membranes (detergent-solubilized-( 125 I)insulin-polyethylene glycol precipitation-sephadex). Proc Natl Acad Sci U S A. 1972 Feb;69(2):318–322. doi: 10.1073/pnas.69.2.318. [DOI] [PMC free article] [PubMed] [Google Scholar]
  12. Faltynek C. R., Princler G. L. Modulation of interferon-alpha and interferon-gamma receptor expression during T-lymphocyte activation and proliferation. J Interferon Res. 1986 Dec;6(6):639–653. doi: 10.1089/jir.1986.6.639. [DOI] [PubMed] [Google Scholar]
  13. Finbloom D. S., Hoover D. L., Wahl L. M. The characteristics of binding of human recombinant interferon-gamma to its receptor on human monocytes and human monocyte-like cell lines. J Immunol. 1985 Jul;135(1):300–305. [PubMed] [Google Scholar]
  14. Green E. D., van Halbeek H., Boime I., Baenziger J. U. Structural elucidation of the disulfated oligosaccharide from bovine lutropin. J Biol Chem. 1985 Dec 15;260(29):15623–15630. [PubMed] [Google Scholar]
  15. Handa K., Suzuki R., Matsui H., Shimizu Y., Kumagai K. Natural killer (NK) cells as a responder to interleukin 2 (IL 2). II. IL 2-induced interferon gamma production. J Immunol. 1983 Feb;130(2):988–992. [PubMed] [Google Scholar]
  16. Laemmli U. K. Cleavage of structural proteins during the assembly of the head of bacteriophage T4. Nature. 1970 Aug 15;227(5259):680–685. doi: 10.1038/227680a0. [DOI] [PubMed] [Google Scholar]
  17. Le J., Prensky W., Yip Y. K., Chang Z., Hoffman T., Stevenson H. C., Balazs I., Sadlik J. R., Vilcek J. Activation of human monocyte cytotoxicity by natural and recombinant immune interferon. J Immunol. 1983 Dec;131(6):2821–2826. [PubMed] [Google Scholar]
  18. Littman S. J., Faltynek C. R., Baglioni C. Binding of human recombinant 125I-interferon gamma to receptors on human cells. J Biol Chem. 1985 Jan 25;260(2):1191–1195. [PubMed] [Google Scholar]
  19. March S. C., Parikh I., Cuatrecasas P. A simplified method for cyanogen bromide activation of agarose for affinity chromatography. Anal Biochem. 1974 Jul;60(1):149–152. doi: 10.1016/0003-2697(74)90139-0. [DOI] [PubMed] [Google Scholar]
  20. Miossec P., Ziff M. Immune interferon enhances the production of interleukin 1 by human endothelial cells stimulated with lipopolysaccharide. J Immunol. 1986 Nov 1;137(9):2848–2852. [PubMed] [Google Scholar]
  21. Molinas F. C., Wietzerbin J., Falcoff E. Human platelets possess receptors for a lymphokine: demonstration of high specific receptors for HuIFN-gamma. J Immunol. 1987 Feb 1;138(3):802–806. [PubMed] [Google Scholar]
  22. Nathan C. F., Murray H. W., Wiebe M. E., Rubin B. Y. Identification of interferon-gamma as the lymphokine that activates human macrophage oxidative metabolism and antimicrobial activity. J Exp Med. 1983 Sep 1;158(3):670–689. doi: 10.1084/jem.158.3.670. [DOI] [PMC free article] [PubMed] [Google Scholar]
  23. Nedwin G. E., Svedersky L. P., Bringman T. S., Palladino M. A., Jr, Goeddel D. V. Effect of interleukin 2, interferon-gamma, and mitogens on the production of tumor necrosis factors alpha and beta. J Immunol. 1985 Oct;135(4):2492–2497. [PubMed] [Google Scholar]
  24. Novick D., Orchansky P., Revel M., Rubinstein M. The human interferon-gamma receptor. Purification, characterization, and preparation of antibodies. J Biol Chem. 1987 Jun 25;262(18):8483–8487. [PubMed] [Google Scholar]
  25. Rashidbaigi A., Kung H. F., Pestka S. Characterization of receptors for immune interferon in U937 cells with 32P-labeled human recombinant immune interferon. J Biol Chem. 1985 Jul 15;260(14):8514–8519. [PubMed] [Google Scholar]
  26. Rosztóczy I., Siroki O., Béládi I. Effects of interferons-alpha, -beta, and -gamma on human interleukin-2 production. J Interferon Res. 1986 Oct;6(5):581–589. doi: 10.1089/jir.1986.6.581. [DOI] [PubMed] [Google Scholar]
  27. Sarkar F. H., Gupta S. L. Receptors for human gamma interferon: binding and crosslinking of 125I-labeled recombinant human gamma interferon to receptors on WISH cells. Proc Natl Acad Sci U S A. 1984 Aug;81(16):5160–5164. doi: 10.1073/pnas.81.16.5160. [DOI] [PMC free article] [PubMed] [Google Scholar]
  28. Schreiber R. D., Pace J. L., Russell S. W., Altman A., Katz D. H. Macrophage-activating factor produced by a T cell hybridoma: physiochemical and biosynthetic resemblance to gamma-interferon. J Immunol. 1983 Aug;131(2):826–832. [PubMed] [Google Scholar]
  29. Sheehan K. C., Calderon J., Schreiber R. D. Generation and characterization of monoclonal antibodies specific for the human IFN-gamma receptor. J Immunol. 1988 Jun 15;140(12):4231–4237. [PubMed] [Google Scholar]
  30. Snapper C. M., Paul W. E. Interferon-gamma and B cell stimulatory factor-1 reciprocally regulate Ig isotype production. Science. 1987 May 22;236(4804):944–947. doi: 10.1126/science.3107127. [DOI] [PubMed] [Google Scholar]
  31. Steeg P. S., Moore R. N., Johnson H. M., Oppenheim J. J. Regulation of murine macrophage Ia antigen expression by a lymphokine with immune interferon activity. J Exp Med. 1982 Dec 1;156(6):1780–1793. doi: 10.1084/jem.156.6.1780. [DOI] [PMC free article] [PubMed] [Google Scholar]
  32. Towbin H., Staehelin T., Gordon J. Electrophoretic transfer of proteins from polyacrylamide gels to nitrocellulose sheets: procedure and some applications. Proc Natl Acad Sci U S A. 1979 Sep;76(9):4350–4354. doi: 10.1073/pnas.76.9.4350. [DOI] [PMC free article] [PubMed] [Google Scholar]
  33. Ucer U., Bartsch H., Scheurich P., Berkovic D., Ertel C., Pfizenmaier K. Quantitation and characterization of gamma-interferon receptors on human tumor cells. Cancer Res. 1986 Oct;46(10):5339–5343. [PubMed] [Google Scholar]
  34. Wheelock E. F. Interferon-like virus-inhibitor induced in human leukocytes by phytohemagglutinin. Science. 1965 Jul 16;149(3681):310–311. [PubMed] [Google Scholar]
  35. Youngner J. S., Salvin S. B. Production and properties of migration inhibitory factor and interferon in the circulation of mice with delayed hypersensitivity. J Immunol. 1973 Dec;111(6):1914–1922. [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