Skip to main content
PMC home page
The Journal of Experimental Medicine logoLink to The Journal of Experimental Medicine
. 1980 Nov 1;152(5):1147–1161. doi: 10.1084/jem.152.5.1147

Structural evidence for distinct IgG subclass-specific Fc receptors on mouse peritoneal macrophages

BC Lane, J Kan-Mitchell, MS Mitchell, SM Cooper
PMCID: PMC2185986  PMID: 7430949

Abstract

Membrane proteins which selectively bind to the Fc portion of IgG were identified in the Nonidet P-40 extracts of radiolabeled thioglycollate- elicited mouse peritoneal macrophages. Affinity columns of various IgG preparations coupled to Sepharose 4B were used to absorb the Fc-binding proteins. Analysis of the acetic acid or sodium dodecyl sulfate (SDS) eluates from aggregated human IgG or antigen-complexed rabbit IgG columns revealed two Fc(gamma)/-specific proteins with apparent 67,000 and 52,000 mol wt. These proteins were not detected in acid or SDS eluates from F(ab’)(2) columns or in eluates from IgG column, over which were passed lysates of Fc receptor-negative cells. With the use of affinity columns that contained aggregated mouse myeloma proteins of different IgG subclasses, we found that the 67,000-dahon protein selectively binds to IgG2a, whereas the 52,000-dalton protein binds to IgG1 and IgG2b. Neither protein was found in SDS eluates from IgG3 columns. Trypsin treatment of the macrophages before detergent lysis removed the 67,000-dalton protein, although it leaves intact the 52,000-dalton protein. These results provide structural confirmation for the existence of separate Fc receptors on mouse macrophages and indicate that the two Fc-binding proteins identified in this study represent all or part of the trypsin- sensitive Fc receptor which binds IgG2a and the trypsin-resistant Fc receptor which binds IgG2b and IgG1.

Full Text

The Full Text of this article is available as a PDF (2.1 MB).

Selected References

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

  1. Anderson C. L., Grey H. M. Physicochemical separation of two distinct Fc receptors on murine macrophage-like cell lines. J Immunol. 1978 Aug;121(2):648–652. [PubMed] [Google Scholar]
  2. Cooper S. M., Sambray Y., Frious G. J. Isolation of separate Fc receptors for IgG complexed to antigen and native IgG from a murine leukaemia. Nature. 1977 Nov 17;270(5634):253–255. doi: 10.1038/270253a0. [DOI] [PubMed] [Google Scholar]
  3. Cooper S. M., Sambray Y. Isolation of a murine leukemia FC receptor by selective release induced by surface redistribution. J Immunol. 1976 Aug;117(2):511–517. [PubMed] [Google Scholar]
  4. Cunningham-Rundles C., Siegal F. P., Good R. A. Isolation and characterization of a human mononuclear cell Fc receptor. Immunochemistry. 1978 Jun;15(6):365–370. doi: 10.1016/0161-5890(78)90132-3. [DOI] [PubMed] [Google Scholar]
  5. D'Urso-Coward M., Cone R. E. Membrane proteins of the P388D1 macrophage cell line: isolation of membrane polypeptides that bind to the Fc portion of aggregated IgG. J Immunol. 1978 Nov;121(5):1973–1980. [PubMed] [Google Scholar]
  6. Diamond B., Birshtein B. K., Scharff M. D. Site of binding of mouse IgG2b to the Fc receptor on mouse macrophages. J Exp Med. 1979 Sep 19;150(3):721–726. doi: 10.1084/jem.150.3.721. [DOI] [PMC free article] [PubMed] [Google Scholar]
  7. Diamond B., Bloom B. R., Scharff M. D. The Fc receptors of primary and cultured phagocytic cells studied with homogeneous antibodies. J Immunol. 1978 Oct;121(4):1329–1333. [PubMed] [Google Scholar]
  8. Dickler H. B. Lymphocyte receptors for immunoglobulin. Adv Immunol. 1976;24:167–214. doi: 10.1016/s0065-2776(08)60330-2. [DOI] [PubMed] [Google Scholar]
  9. Frade R., Kourilsky F. M. Preliminary characterization of a glycoprotein having Fc receptor properties extracted from a T cell lymphoma (L-5178-Y). Eur J Immunol. 1977 Sep;7(9):663–666. doi: 10.1002/eji.1830070916. [DOI] [PubMed] [Google Scholar]
  10. Haeffner-Cavaillon N., Klein M., Dorrington K. J. Studies on the Fc gamma receptor of the murine macrophage-like cell line P388D1. I. The binding of homologous and heterologous immunoglobulin G1. J Immunol. 1979 Nov;123(5):1905–1913. [PubMed] [Google Scholar]
  11. Heusser C. H., Anderson C. L., Grey H. M. Receptors for IgG: subclass specificity of receptors on different mouse cell types and the definition of two distinct receptors on a macrophage cell line. J Exp Med. 1977 May 1;145(5):1316–1327. doi: 10.1084/jem.145.5.1316. [DOI] [PMC free article] [PubMed] [Google Scholar]
  12. Kulczycki A., Jr, Krause V., Killion C. C., Atkinson J. P. Purification of Fc gamma receptor from rabbit alveolar macrophages that retains ligand-binding activity. J Immunol. 1980 Jun;124(6):2772–2779. [PubMed] [Google Scholar]
  13. Kulczycki A., Jr, Metzger H. The interaction of IgE with rat basophilic leukemia cells. II. Quantitative aspects of the binding reaction. J Exp Med. 1974 Dec 1;140(6):1676–1695. doi: 10.1084/jem.140.6.1676. [DOI] [PMC free article] [PubMed] [Google Scholar]
  14. Kulczycki A., Jr, Parker C. W. The cell surface receptor for immunoglobulin E. I. The use of repetitive affinity chromatography for the purification of a mammalian receptor. J Biol Chem. 1979 May 10;254(9):3187–3193. [PubMed] [Google Scholar]
  15. 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]
  16. Laskey R. A., Mills A. D. Enhanced autoradiographic detection of 32P and 125I using intensifying screens and hypersensitized film. FEBS Lett. 1977 Oct 15;82(2):314–316. doi: 10.1016/0014-5793(77)80609-1. [DOI] [PubMed] [Google Scholar]
  17. Loube S. R., McNabb T. C., Dorrington K. J. Isolation of an Fcgamma-binding protein from the cell membrane of a macrophage-like cell line (P388D1) after detergent solubilization. J Immunol. 1978 Mar;120(3):709–715. [PubMed] [Google Scholar]
  18. MacLennan I. C. Antibody in the induction and inhibition of lymphocyte cytotoxicity. Transplant Rev. 1972;13:67–90. doi: 10.1111/j.1600-065x.1972.tb00060.x. [DOI] [PubMed] [Google Scholar]
  19. Markwell M. A., Fox C. F. Surface-specific iodination of membrane proteins of viruses and eucaryotic cells using 1,3,4,6-tetrachloro-3alpha,6alpha-diphenylglycoluril. Biochemistry. 1978 Oct 31;17(22):4807–4817. doi: 10.1021/bi00615a031. [DOI] [PubMed] [Google Scholar]
  20. Molenaar J. L., van Galen M., Hannema A. J., Zeijlemaker W., Pondman K. W. Spontaneous release of Fc receptor-like material from human lymphoblastoid cell lines. Eur J Immunol. 1977 Apr;7(4):230–236. doi: 10.1002/eji.1830070409. [DOI] [PubMed] [Google Scholar]
  21. O'Farrell P. H. High resolution two-dimensional electrophoresis of proteins. J Biol Chem. 1975 May 25;250(10):4007–4021. [PMC free article] [PubMed] [Google Scholar]
  22. Rask L., Klarkeskog L., Ostberg L., Peterson P. A. Isolation and properties of a murine spleen cell Fc receptor. Nature. 1975 Sep 18;257(5523):231–233. doi: 10.1038/257231a0. [DOI] [PubMed] [Google Scholar]
  23. Silverstein S. C., Steinman R. M., Cohn Z. A. Endocytosis. Annu Rev Biochem. 1977;46:669–722. doi: 10.1146/annurev.bi.46.070177.003321. [DOI] [PubMed] [Google Scholar]
  24. Thoenes J., Stein H. Properties of an Fc gamma-binding protein isolated from human leukemic B cells. J Exp Med. 1979 Nov 1;150(5):1049–1066. doi: 10.1084/jem.150.5.1049. [DOI] [PMC free article] [PubMed] [Google Scholar]
  25. Unkeless J. C. Characterization of a monoclonal antibody directed against mouse macrophage and lymphocyte Fc receptors. J Exp Med. 1979 Sep 19;150(3):580–596. doi: 10.1084/jem.150.3.580. [DOI] [PMC free article] [PubMed] [Google Scholar]
  26. Unkeless J. C., Eisen H. N. Binding of monomeric immunoglobulins to Fc receptors of mouse macrophages. J Exp Med. 1975 Dec 1;142(6):1520–1533. doi: 10.1084/jem.142.6.1520. [DOI] [PMC free article] [PubMed] [Google Scholar]
  27. Unkeless J. C., Kaplan G., Plutner H., Cohn Z. A. Fc-receptor variants of a mouse macrophage cell line. Proc Natl Acad Sci U S A. 1979 Mar;76(3):1400–1404. doi: 10.1073/pnas.76.3.1400. [DOI] [PMC free article] [PubMed] [Google Scholar]
  28. Unkeless J. C. The presence of two Fc receptors on mouse macrophages: evidence from a variant cell line and differential trypsin sensitivity. J Exp Med. 1977 Apr 1;145(4):931–945. doi: 10.1084/jem.145.4.931. [DOI] [PMC free article] [PubMed] [Google Scholar]
  29. Walker W. S. Mediation of macrophage cytolytic and phagocytic activities by antibodies of different classes and class-specific Fc-receptors. J Immunol. 1977 Aug;119(2):367–373. [PubMed] [Google Scholar]
  30. Walker W. S. Separate Fc-receptors for immunoglogulins IgG2a and IgG2b on an established cell line of mouse macrophages. J Immunol. 1976 Apr;116(4):911–914. [PubMed] [Google Scholar]
  31. Yagawa K., Onoue K., Aida Y. Structural studies of Fc receptors. I. Binding properties, solubilization, and partial characterization of fc receptors of macrophages. J Immunol. 1979 Jan;122(1):366–373. [PubMed] [Google Scholar]
  32. Yam L. T., Li C. Y., Crosby W. H. Cytochemical identification of monocytes and granulocytes. Am J Clin Pathol. 1971 Mar;55(3):283–290. doi: 10.1093/ajcp/55.3.283. [DOI] [PubMed] [Google Scholar]
  33. Zeligs B. J., Nerurkar L. S., Bellanti J. A. Maturation of the rabbit alveolar macrophage during animal development. III. Phagocytic and bactericidal functions. Pediatr Res. 1977 Dec;11(12):1208–1211. doi: 10.1203/00006450-197712000-00008. [DOI] [PubMed] [Google Scholar]

Articles from The Journal of Experimental Medicine are provided here courtesy of The Rockefeller University Press

RESOURCES