Skip to main content
PMC home page
Biophysical Journal logoLink to Biophysical Journal
. 1992 May;61(5):1402–1412. doi: 10.1016/S0006-3495(92)81946-X

Effects of protein concentration on IgE receptor mobility in rat basophilic leukemia cell plasma membranes.

J L Thomas 1, T J Feder 1, W W Webb 1
PMCID: PMC1260401  PMID: 1534697

Abstract

The ability of variations of membrane protein concentrations to modulate the lateral diffusion rate of an exemplary membrane protein has been studied in healthy and osmotically shocked cultured cells of the rat basophilic leukemia cell line, 2H3 subclone. Cell surface protein was redistributed by the method of in situ electrophoresis; exposure to electric fields of 1.25-5 V/cm results in cathodal migration of the majority of the surface proteins on this cell type (Ryan, T. A., J. Myers, D. Holowka, B. Baird, and W. W. Webb. Science [Wash. DC]. 239:61-64). Even in these small fields, the steady-state distribution becomes "crowded" with more than an 80% protein occupancy of accessible membrane area at the cathodal end of these spheroidal cells, and the anodal end becomes significantly depleted. We have employed fringe pattern fluorescence photobleaching with CCD imaging detection to measure lateral diffusion coefficients of the liganded IgE receptor on both crowded and uncrowded regions of individual rat basophilic leukemia cells. We find no significant difference in lateral diffusion rates in these regions. Cells swollen by hypoosmotic stress exhibit faster diffusion overall, with the uncrowded regions having a significantly greater increase in diffusion coefficient than the crowded regions. These results are consistent with the partial or total release of cytoskeletal constraints to membrane protein diffusion induced by osmotic stress.

Full text

PDF
1402

Images in this article

1405FIGURE 2
on p.1405
1408FIGURE 7
on p.1408

Selected References

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

  1. Apgar J. R. Antigen-induced cross-linking of the IgE receptor leads to an association with the detergent-insoluble membrane skeleton of rat basophilic leukemia (RBL-2H3) cells. J Immunol. 1990 Dec 1;145(11):3814–3822. [PubMed] [Google Scholar]
  2. Apgar J. R., Mescher M. F. Agorins: major structural proteins of the plasma membrane skeleton of P815 tumor cells. J Cell Biol. 1986 Aug;103(2):351–360. doi: 10.1083/jcb.103.2.351. [DOI] [PMC free article] [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. Davoust J., Devaux P. F., Leger L. Fringe pattern photobleaching, a new method for the measurement of transport coefficients of biological macromolecules. EMBO J. 1982;1(10):1233–1238. doi: 10.1002/j.1460-2075.1982.tb00018.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  5. Golan D. E., Alecio M. R., Veatch W. R., Rando R. R. Lateral mobility of phospholipid and cholesterol in the human erythrocyte membrane: effects of protein-lipid interactions. Biochemistry. 1984 Jan 17;23(2):332–339. doi: 10.1021/bi00297a024. [DOI] [PubMed] [Google Scholar]
  6. Jaffe L. F. Electrophoresis along cell membranes. Nature. 1977 Feb 17;265(5595):600–602. doi: 10.1038/265600a0. [DOI] [PubMed] [Google Scholar]
  7. McCloskey M. A., Liu Z. Y., Poo M. M. Lateral electromigration and diffusion of Fc epsilon receptors on rat basophilic leukemia cells: effects of IgE binding. J Cell Biol. 1984 Sep;99(3):778–787. doi: 10.1083/jcb.99.3.778. [DOI] [PMC free article] [PubMed] [Google Scholar]
  8. 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]
  9. Menon A. K., Holowka D., Webb W. W., Baird B. Clustering, mobility, and triggering activity of small oligomers of immunoglobulin E on rat basophilic leukemia cells. J Cell Biol. 1986 Feb;102(2):534–540. doi: 10.1083/jcb.102.2.534. [DOI] [PMC free article] [PubMed] [Google Scholar]
  10. Menon A. K., Holowka D., Webb W. W., Baird B. Cross-linking of receptor-bound IgE to aggregates larger than dimers leads to rapid immobilization. J Cell Biol. 1986 Feb;102(2):541–550. doi: 10.1083/jcb.102.2.541. [DOI] [PMC free article] [PubMed] [Google Scholar]
  11. Mescher M. F., Jose M. J., Balk S. P. Actin-containing matrix associated with the plasma membrane of murine tumour and lymphoid cells. Nature. 1981 Jan 15;289(5794):139–144. doi: 10.1038/289139a0. [DOI] [PubMed] [Google Scholar]
  12. Okada C. Y., Rechsteiner M. Introduction of macromolecules into cultured mammalian cells by osmotic lysis of pinocytic vesicles. Cell. 1982 May;29(1):33–41. doi: 10.1016/0092-8674(82)90087-3. [DOI] [PubMed] [Google Scholar]
  13. Peters R., Cherry R. J. Lateral and rotational diffusion of bacteriorhodopsin in lipid bilayers: experimental test of the Saffman-Delbrück equations. Proc Natl Acad Sci U S A. 1982 Jul;79(14):4317–4321. doi: 10.1073/pnas.79.14.4317. [DOI] [PMC free article] [PubMed] [Google Scholar]
  14. Pfeiffer J. R., Seagrave J. C., Davis B. H., Deanin G. G., Oliver J. M. Membrane and cytoskeletal changes associated with IgE-mediated serotonin release from rat basophilic leukemia cells. J Cell Biol. 1985 Dec;101(6):2145–2155. doi: 10.1083/jcb.101.6.2145. [DOI] [PMC free article] [PubMed] [Google Scholar]
  15. Poo M., Lam J. W., Orida N., Chao A. W. Electrophoresis and diffusion in the plane of the cell membrane. Biophys J. 1979 Apr;26(1):1–21. doi: 10.1016/S0006-3495(79)85231-5. [DOI] [PMC free article] [PubMed] [Google Scholar]
  16. Quinn P., Griffiths G., Warren G. Density of newly synthesized plasma membrane proteins in intracellular membranes II. Biochemical studies. J Cell Biol. 1984 Jun;98(6):2142–2147. doi: 10.1083/jcb.98.6.2142. [DOI] [PMC free article] [PubMed] [Google Scholar]
  17. Ryan T. A., Myers J., Holowka D., Baird B., Webb W. W. Molecular crowding on the cell surface. Science. 1988 Jan 1;239(4835):61–64. doi: 10.1126/science.2962287. [DOI] [PubMed] [Google Scholar]
  18. 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]
  19. Saxton M. J. Lateral diffusion in an archipelago. Distance dependence of the diffusion coefficient. Biophys J. 1989 Sep;56(3):615–622. doi: 10.1016/S0006-3495(89)82708-0. [DOI] [PMC free article] [PubMed] [Google Scholar]
  20. Saxton M. J. Lateral diffusion in an archipelago. The effect of mobile obstacles. Biophys J. 1987 Dec;52(6):989–997. doi: 10.1016/S0006-3495(87)83291-5. [DOI] [PMC free article] [PubMed] [Google Scholar]
  21. Schlessinger J., Koppel D. E., Axelrod D., Jacobson K., Webb W. W., Elson E. L. Lateral transport on cell membranes: mobility of concanavalin A receptors on myoblasts. Proc Natl Acad Sci U S A. 1976 Jul;73(7):2409–2413. doi: 10.1073/pnas.73.7.2409. [DOI] [PMC free article] [PubMed] [Google Scholar]
  22. Tank D. W., Wu E. S., Meers P. R., Webb W. W. Lateral diffusion of gramicidin C in phospholipid multibilayers. Effects of cholesterol and high gramicidin concentration. Biophys J. 1982 Nov;40(2):129–135. doi: 10.1016/S0006-3495(82)84467-6. [DOI] [PMC free article] [PubMed] [Google Scholar]
  23. Tank D. W., Wu E. S., Webb W. W. Enhanced molecular diffusibility in muscle membrane blebs: release of lateral constraints. J Cell Biol. 1982 Jan;92(1):207–212. doi: 10.1083/jcb.92.1.207. [DOI] [PMC free article] [PubMed] [Google Scholar]
  24. Webb W. W., Barak L. S., Tank D. W., Wu E. S. Molecular mobility on the cell surface. Biochem Soc Symp. 1981;(46):191–205. [PubMed] [Google Scholar]
  25. Wu E. S., Tank D. W., Webb W. W. Unconstrained lateral diffusion of concanavalin A receptors on bulbous lymphocytes. Proc Natl Acad Sci U S A. 1982 Aug;79(16):4962–4966. doi: 10.1073/pnas.79.16.4962. [DOI] [PMC free article] [PubMed] [Google Scholar]
  26. Yechiel E., Edidin M. Micrometer-scale domains in fibroblast plasma membranes. J Cell Biol. 1987 Aug;105(2):755–760. doi: 10.1083/jcb.105.2.755. [DOI] [PMC free article] [PubMed] [Google Scholar]

Articles from Biophysical Journal are provided here courtesy of The Biophysical Society

RESOURCES