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. 1984 Sep 1;99(3):778–787. doi: 10.1083/jcb.99.3.778

Lateral electromigration and diffusion of Fc epsilon receptors on rat basophilic leukemia cells: effects of IgE binding

PMCID: PMC2113395  PMID: 6236227

Abstract

We have used in situ electromigration and post-field relaxation (Poo, M.-m., 1981, Annu. Rev. Biophys. Bioeng., 10:245-276) to assess the effect of immunoglobulin E (IgE) binding on the lateral mobility of IgE- Fc receptors in the plasmalemma of rat basophilic leukemia (RBL) cells. Bound IgE sharply increased the receptor's electrokinetic mobility, whereas removal of cell surface neuraminic acids cut it to near zero. In contrast, we found only a small difference between the lateral diffusion coefficients (D) of vacant and IgE-occupied Fc receptors (D: 4 vs. 3 X 10(-10) cm2/s at 24 degrees C). This is true for monomeric rat IgE; with mouse IgE, the difference in apparent diffusion rates was slightly greater (D: 4.5 vs. 2.3 X 10(-10) cm2/s at 24 degrees C). This range of D values is close to that found in previous photobleaching studies of the IgE-Fc epsilon receptor complex in RBL cells and rat mast cells. Moreover, enzymatic depletion of cell coat components did not measurably alter the diffusion rate of IgE-occupied receptors. Thus, binding of fluorescent macromolecular probes to cell surface proteins need not severely impede lateral diffusion of the probed species. If the glycocalyx of RBL cells does limit lateral diffusion of the Fc epsilon receptor, it must act primarily on the receptor itself, rather than on receptor-bound IgE.

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Selected References

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  1. Amante L., Ancona A., Forni L. The conjugation of immunoglobulins with tetramethylrhodamine isothiocyanate. A comparison between the amorphous and the crystalline fluorochrome. J Immunol Methods. 1972 May;1(3):289–301. doi: 10.1016/0022-1759(72)90006-3. [DOI] [PubMed] [Google Scholar]
  2. Axelrod D., Wight A., Webb W., Horwitz A. Influence of membrane lipids on acetylcholine receptor and lipid probe diffusion in cultured myotube membrane. Biochemistry. 1978 Aug 22;17(17):3604–3609. doi: 10.1021/bi00610a029. [DOI] [PubMed] [Google Scholar]
  3. Barak L. S., Webb W. W. Diffusion of low density lipoprotein-receptor complex on human fibroblasts. J Cell Biol. 1982 Dec;95(3):846–852. doi: 10.1083/jcb.95.3.846. [DOI] [PMC free article] [PubMed] [Google Scholar]
  4. Barsumian E. L., Isersky C., Petrino M. G., Siraganian R. P. IgE-induced histamine release from rat basophilic leukemia cell lines: isolation of releasing and nonreleasing clones. Eur J Immunol. 1981 Apr;11(4):317–323. doi: 10.1002/eji.1830110410. [DOI] [PubMed] [Google Scholar]
  5. Bazin H., Querinjean P., Beckers A., Heremans J. F., Dessy F. Transplantable immunoglobulin-secreting tumours in rats. IV. Sixty-three IgE-secreting immunocytoma tumours. Immunology. 1974 Apr;26(4):713–723. [PMC free article] [PubMed] [Google Scholar]
  6. Braun J., Hochman P. S., Unanue E. R. Ligand-induced association of surface immunoglobulin with the detergent-insoluble cytoskeletal matrix of the B lymphocyte. J Immunol. 1982 Mar;128(3):1198–1204. [PubMed] [Google Scholar]
  7. Conrad D. H., Bazin H., Sehon A. H., Froese A. Binding parameters of the interaction between rat IgE and rat mast cell receptors. J Immunol. 1975 Jun;114(6):1688–1691. [PubMed] [Google Scholar]
  8. Conrad D. H., Studer E., Gervasoni J., Mohanakumar T. Properties of two monoclonal antibodies directed against the Fc and Fab' regions of rat IgE. Int Arch Allergy Appl Immunol. 1983;70(4):352–360. doi: 10.1159/000233347. [DOI] [PubMed] [Google Scholar]
  9. Edidin M., Wei T. Lateral diffusion of H-2 antigens on mouse fibroblasts. J Cell Biol. 1982 Nov;95(2 Pt 1):458–462. doi: 10.1083/jcb.95.2.458. [DOI] [PMC free article] [PubMed] [Google Scholar]
  10. Goldberg R. L., Toole B. P. Hyaluronate coat formation and cell spreading in rat fibrosarcoma cells. Exp Cell Res. 1984 Mar;151(1):258–263. doi: 10.1016/0014-4827(84)90374-4. [DOI] [PubMed] [Google Scholar]
  11. Hafeman D. G., Smith L. M., Fearon D. T., McConnell H. M. Lipid monolayer-coated solid surfaces do not perturb the lateral motion and distribution of C3b receptors on neutrophils. J Cell Biol. 1982 Jul;94(1):224–227. doi: 10.1083/jcb.94.1.224. [DOI] [PMC free article] [PubMed] [Google Scholar]
  12. Hafeman D. G., von Tscharner V., McConnell H. M. Specific antibody-dependent interactions between macrophages and lipid haptens in planar lipid monolayers. Proc Natl Acad Sci U S A. 1981 Jul;78(7):4552–4556. doi: 10.1073/pnas.78.7.4552. [DOI] [PMC free article] [PubMed] [Google Scholar]
  13. Horsfield G. I. The effect of compound 48/80 on the rat mast cell. J Pathol Bacteriol. 1965 Oct;90(2):599–605. doi: 10.1002/path.1700900228. [DOI] [PubMed] [Google Scholar]
  14. Lawson D., Fewtrell C., Raff M. C. Localized mast cell degranulation induced by concanavalin A-sepharose beads. Implications for the Ca2+ hypothesis of stimulus-secretion coupling. J Cell Biol. 1978 Nov;79(2 Pt 1):394–400. doi: 10.1083/jcb.79.2.394. [DOI] [PMC free article] [PubMed] [Google Scholar]
  15. Liu F. T., Bohn J. W., Ferry E. L., Yamamoto H., Molinaro C. A., Sherman L. A., Klinman N. R., Katz D. H. Monoclonal dinitrophenyl-specific murine IgE antibody: preparation, isolation, and characterization. J Immunol. 1980 Jun;124(6):2728–2737. [PubMed] [Google Scholar]
  16. Maxfield F. R., Schlessinger J., Shechter Y., Pastan I., Willingham M. C. Collection of insulin, EGF and alpha2-macroglobulin in the same patches on the surface of cultured fibroblasts and common internalization. Cell. 1978 Aug;14(4):805–810. doi: 10.1016/0092-8674(78)90336-7. [DOI] [PMC free article] [PubMed] [Google Scholar]
  17. McLaughlin S., Poo M. M. The role of electro-osmosis in the electric-field-induced movement of charged macromolecules on the surfaces of cells. Biophys J. 1981 Apr;34(1):85–93. doi: 10.1016/S0006-3495(81)84838-2. [DOI] [PMC free article] [PubMed] [Google Scholar]
  18. Metzger H., Budman D., Lucky P. Interaction of IgE with rat basophilic leukemia cells--V. Binding properties of cell free particles. Immunochemistry. 1976 May;13(5):417–423. doi: 10.1016/0019-2791(76)90377-3. [DOI] [PubMed] [Google Scholar]
  19. Meyer C., Wahl L. M., Stadler B. M., Siraganian R. P. Cell cycle associated changes in histamine release from rat basophilic leukemia cells separated by counterflow centrifugal elutriation. J Immunol. 1983 Aug;131(2):911–914. [PubMed] [Google Scholar]
  20. Poo M. In situ electrophoresis of membrane components. Annu Rev Biophys Bioeng. 1981;10:245–276. doi: 10.1146/annurev.bb.10.060181.001333. [DOI] [PubMed] [Google Scholar]
  21. Saffman P. G., Delbrück M. Brownian motion in biological membranes. Proc Natl Acad Sci U S A. 1975 Aug;72(8):3111–3113. doi: 10.1073/pnas.72.8.3111. [DOI] [PMC free article] [PubMed] [Google Scholar]
  22. Schlessinger J., Webb W. W., Elson E. L., Metzger H. Lateral motion and valence of Fc receptors on rat peritoneal mast cells. Nature. 1976 Dec 9;264(5586):550–552. doi: 10.1038/264550a0. [DOI] [PubMed] [Google Scholar]
  23. Segal D. M., Hurwitz E. Dimers and trimers of immunoglobulin G covalently cross-linked with a bivalent affinity label. Biochemistry. 1976 Nov 30;15(24):5253–5258. doi: 10.1021/bi00669a009. [DOI] [PubMed] [Google Scholar]
  24. Smith L. M., Parce J. W., Smith B. A., McConnell H. M. Antibodies bound to lipid haptens in model membranes diffuse as rapidly as the lipids themselves. Proc Natl Acad Sci U S A. 1979 Sep;76(9):4177–4179. doi: 10.1073/pnas.76.9.4177. [DOI] [PMC free article] [PubMed] [Google Scholar]
  25. Smith L. M., Petty H. R., Parham P., McConnell H. M. Cell surface properties of HLA antigens on Epstein-Barr virus-transformed cell lines. Proc Natl Acad Sci U S A. 1982 Jan;79(2):608–612. doi: 10.1073/pnas.79.2.608. [DOI] [PMC free article] [PubMed] [Google Scholar]
  26. Taurog J. D., Fewtrell C., Becker E. L. IgE mediated triggering of rat basophil leukemia cells: lack of evidence for serine esterase activation. J Immunol. 1979 Jun;122(6):2150–2153. [PubMed] [Google Scholar]
  27. Titus J. A., Haugland R., Sharrow S. O., Segal D. M. Texas Red, a hydrophilic, red-emitting fluorophore for use with fluorescein in dual parameter flow microfluorometric and fluorescence microscopic studies. J Immunol Methods. 1982;50(2):193–204. doi: 10.1016/0022-1759(82)90225-3. [DOI] [PubMed] [Google Scholar]
  28. Wank S. A., DeLisi C., Metzger H. Analysis of the rate-limiting step in a ligand-cell receptor interaction: the immunoglobulin E system. Biochemistry. 1983 Feb 15;22(4):954–959. doi: 10.1021/bi00273a038. [DOI] [PubMed] [Google Scholar]
  29. Wolf D. E., Edidin M., Dragsten P. R. Effect of bleaching light on measurements of lateral diffusion in cell membranes by the fluorescence photobleaching recovery method. Proc Natl Acad Sci U S A. 1980 Apr;77(4):2043–2045. doi: 10.1073/pnas.77.4.2043. [DOI] [PMC free article] [PubMed] [Google Scholar]
  30. Zagyansky Y. A., Jard S. Does lectin-receptor complex formation produce zones of restricted mobility within the membrane? Nature. 1979 Aug 16;280(5723):591–593. doi: 10.1038/280591a0. [DOI] [PubMed] [Google Scholar]

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