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. 1996 Jun;70(6):2767–2773. doi: 10.1016/S0006-3495(96)79846-6

Constrained diffusion or immobile fraction on cell surfaces: a new interpretation.

T J Feder 1, I Brust-Mascher 1, J P Slattery 1, B Baird 1, W W Webb 1
PMCID: PMC1225256  PMID: 8744314

Abstract

Protein lateral mobility in cell membranes is generally measured using fluorescence photobleaching recovery (FPR). Since the development of this technique, the data have been interpreted by assuming free Brownian diffusion of cell surface receptors in two dimensions, an interpretation that requires that a subset of the diffusing species remains immobile. The origin of this so-called immobile fraction remains a mystery. In FPR, the motions of thousands of particles are inherently averaged, inevitably masking the details of individual motions. Recently, tracking of individual cell surface receptors has identified several distinct types of motion (Gross and Webb, 1988; Ghosh and Webb, 1988, 1990, 1994; Kusumi et al. 1993; Qian et al. 1991; Slattery, 1995), thereby calling into question the classical interpretation of FPR data as free Brownian motion of a limited mobile fraction. We have measured the motion of fluorescently labeled immunoglobulin E complexed to high affinity receptors (Fc epsilon RI) on rat basophilic leukemia cells using both single particle tracking and FPR. As in previous studies, our tracking results show that individual receptors may diffuse freely, or may exhibit restricted, time-dependent (anomalous) diffusion. Accordingly, we have analyzed FPR data by a new model to take this varied motion into account, and we show that the immobile fraction may be due to particles moving with the anomalous subdiffusion associated with restricted lateral mobility. Anomalous subdiffusion denotes random molecular motion in which the mean square displacements grow as a power law in time with a fractional positive exponent less than one. These findings call for a new model of cell membrane structure.

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Selected References

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  1. Anderson C. M., Georgiou G. N., Morrison I. E., Stevenson G. V., Cherry R. J. Tracking of cell surface receptors by fluorescence digital imaging microscopy using a charge-coupled device camera. Low-density lipoprotein and influenza virus receptor mobility at 4 degrees C. J Cell Sci. 1992 Feb;101(Pt 2):415–425. doi: 10.1242/jcs.101.2.415. [DOI] [PubMed] [Google Scholar]
  2. Axelrod D., Koppel D. E., Schlessinger J., Elson E., Webb W. W. Mobility measurement by analysis of fluorescence photobleaching recovery kinetics. Biophys J. 1976 Sep;16(9):1055–1069. doi: 10.1016/S0006-3495(76)85755-4. [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. 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. Edidin M., Stroynowski I. Differences between the lateral organization of conventional and inositol phospholipid-anchored membrane proteins. A further definition of micrometer scale membrane domains. J Cell Biol. 1991 Mar;112(6):1143–1150. doi: 10.1083/jcb.112.6.1143. [DOI] [PMC free article] [PubMed] [Google Scholar]
  6. Erickson J., Kane P., Goldstein B., Holowka D., Baird B. Cross-linking of IgE-receptor complexes at the cell surface: a fluorescence method for studying the binding of monovalent and bivalent haptens to IgE. Mol Immunol. 1986 Jul;23(7):769–781. doi: 10.1016/0161-5890(86)90089-1. [DOI] [PubMed] [Google Scholar]
  7. Ghosh R. N., Webb W. W. Automated detection and tracking of individual and clustered cell surface low density lipoprotein receptor molecules. Biophys J. 1994 May;66(5):1301–1318. doi: 10.1016/S0006-3495(94)80939-7. [DOI] [PMC free article] [PubMed] [Google Scholar]
  8. Jacobson K., Sheets E. D., Simson R. Revisiting the fluid mosaic model of membranes. Science. 1995 Jun 9;268(5216):1441–1442. doi: 10.1126/science.7770769. [DOI] [PubMed] [Google Scholar]
  9. Kucik D. F., Elson E. L., Sheetz M. P. Forward transport of glycoproteins on leading lamellipodia in locomoting cells. Nature. 1989 Jul 27;340(6231):315–317. doi: 10.1038/340315a0. [DOI] [PubMed] [Google Scholar]
  10. 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]
  11. Kusumi A., Sako Y., Yamamoto M. Confined lateral diffusion of membrane receptors as studied by single particle tracking (nanovid microscopy). Effects of calcium-induced differentiation in cultured epithelial cells. Biophys J. 1993 Nov;65(5):2021–2040. doi: 10.1016/S0006-3495(93)81253-0. [DOI] [PMC free article] [PubMed] [Google Scholar]
  12. Lee G. M., Zhang F., Ishihara A., McNeil C. L., Jacobson K. A. Unconfined lateral diffusion and an estimate of pericellular matrix viscosity revealed by measuring the mobility of gold-tagged lipids. J Cell Biol. 1993 Jan;120(1):25–35. doi: 10.1083/jcb.120.1.25. [DOI] [PMC free article] [PubMed] [Google Scholar]
  13. Nagle J. F. Long tail kinetics in biophysics? Biophys J. 1992 Aug;63(2):366–370. doi: 10.1016/S0006-3495(92)81602-8. [DOI] [PMC free article] [PubMed] [Google Scholar]
  14. 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]
  15. Qian H., Sheetz M. P., Elson E. L. Single particle tracking. Analysis of diffusion and flow in two-dimensional systems. Biophys J. 1991 Oct;60(4):910–921. doi: 10.1016/S0006-3495(91)82125-7. [DOI] [PMC free article] [PubMed] [Google Scholar]
  16. 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]
  17. Sheetz M. P., Turney S., Qian H., Elson E. L. Nanometre-level analysis demonstrates that lipid flow does not drive membrane glycoprotein movements. Nature. 1989 Jul 27;340(6231):284–288. doi: 10.1038/340284a0. [DOI] [PubMed] [Google Scholar]
  18. Singer S. J., Nicolson G. L. The fluid mosaic model of the structure of cell membranes. Science. 1972 Feb 18;175(4023):720–731. doi: 10.1126/science.175.4023.720. [DOI] [PubMed] [Google Scholar]
  19. Vaz W. L., Criado M., Madeira V. M., Schoellmann G., Jovin T. M. Size dependence of the translational diffusion of large integral membrane proteins in liquid-crystalline phase lipid bilayers. A study using fluorescence recovery after photobleaching. Biochemistry. 1982 Oct 26;21(22):5608–5612. doi: 10.1021/bi00265a034. [DOI] [PubMed] [Google Scholar]
  20. 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]
  21. Yguerabide J., Schmidt J. A., Yguerabide E. E. Lateral mobility in membranes as detected by fluorescence recovery after photobleaching. Biophys J. 1982 Oct;40(1):69–75. doi: 10.1016/S0006-3495(82)84459-7. [DOI] [PMC free article] [PubMed] [Google Scholar]
  22. Zhang F., Lee G. M., Jacobson K. Protein lateral mobility as a reflection of membrane microstructure. Bioessays. 1993 Sep;15(9):579–588. doi: 10.1002/bies.950150903. [DOI] [PubMed] [Google Scholar]
  23. de Brabander M., Nuydens R., Ishihara A., Holifield B., Jacobson K., Geerts H. Lateral diffusion and retrograde movements of individual cell surface components on single motile cells observed with Nanovid microscopy. J Cell Biol. 1991 Jan;112(1):111–124. doi: 10.1083/jcb.112.1.111. [DOI] [PMC free article] [PubMed] [Google Scholar]

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