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. 1991 Jul 15;88(14):6274–6278. doi: 10.1073/pnas.88.14.6274

Direct observation of brownian motion of lipids in a membrane.

G M Lee 1, A Ishihara 1, K A Jacobson 1
PMCID: PMC52065  PMID: 1712486

Abstract

Nanovid microscopy, which uses 30- to 40-nm colloidal gold probes combined with video-enhanced contrast, can be used to examine random and directed movements of individual molecules in the plasma membrane of living cells. To validate the technique in a model system, the movements of lipid molecules were followed in a supported, planar bilayer containing fluorescein-conjugated phosphatidylethanolamine (Fl-PtdEtn) labeled with 30-nm gold anti-fluorescein (anti-Fl). Multivalent gold probes were prepared by conjugating only anti-Fl to the gold. Paucivalent probes were prepared by mixing an irrelevant antibody with the anti-Fl prior to conjugation. The membrane-bound gold particles moved in random patterns that were indistinguishable from those produced by computer simulations of two-dimensional random motion. The multivalent gold probes had an average lateral diffusion coefficient (D) of 0.26 x 10(-8) cm2/sec, and paucivalent probes had an average D of 0.73 x 10(-8) cm2/sec. Sixteen percent of the multivalent and 50% of the paucivalent probes had values for D in excess of 0.6 x 10(-8) cm2/sec, which, after allowance for stochastic variation, are consistent with the D of 1.3 x 10(-8) cm2/sec measured by fluorescence recovery after photobleaching of Fl-PtdEtn in the planar bilayer. The effect of valency on diffusion suggests that the multivalent gold binds several lipids forming a disk up to 30-40 nm in diameter, resulting in reduced diffusion with respect to the paucivalent gold, which binds one or a very few lipids. Provided the valency of the gold probe is considered in the interpretation of the results. Nanovid microscopy is a valid method for analyzing the movements of single or small groups of molecules within membranes.

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Selected References

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

  1. Billings-Gagliardi S., Pockwinse S. M., Schneider G. B. Surface coats on human lymphocytes: freeze-drying and staining with cations. Am J Anat. 1979 Feb;154(2):267–276. doi: 10.1002/aja.1001540210. [DOI] [PubMed] [Google Scholar]
  2. De Brabander M., Nuydens R., Geerts H., Hopkins C. R. Dynamic behavior of the transferrin receptor followed in living epidermoid carcinoma (A431) cells with nanovid microscopy. Cell Motil Cytoskeleton. 1988;9(1):30–47. doi: 10.1002/cm.970090105. [DOI] [PubMed] [Google Scholar]
  3. De Roe C., Courtoy P. J., Baudhuin P. A model of protein-colloidal gold interactions. J Histochem Cytochem. 1987 Nov;35(11):1191–1198. doi: 10.1177/35.11.3655323. [DOI] [PubMed] [Google Scholar]
  4. Ishihara A., Holifield B., Jacobson K. Analysis of lateral redistribution of a plasma membrane glycoprotein-monoclonal antibody complex [corrected]. J Cell Biol. 1988 Feb;106(2):329–343. doi: 10.1083/jcb.106.2.329. [DOI] [PMC free article] [PubMed] [Google Scholar]
  5. Ishihara A., Hou Y., Jacobson K. The Thy-1 antigen exhibits rapid lateral diffusion in the plasma membrane of rodent lymphoid cells and fibroblasts. Proc Natl Acad Sci U S A. 1987 Mar;84(5):1290–1293. doi: 10.1073/pnas.84.5.1290. [DOI] [PMC free article] [PubMed] [Google Scholar]
  6. Jacobson K., Derzko Z., Wu E. S., Hou Y., Poste G. Measurement of the lateral mobility of cell surface components in single, living cells by fluorescence recovery after photobleaching. J Supramol Struct. 1976;5(4):565(417)–576(428). doi: 10.1002/jss.400050411. [DOI] [PubMed] [Google Scholar]
  7. Kucik D. F., Elson E. L., Sheetz M. P. Cell migration does not produce membrane flow. J Cell Biol. 1990 Oct;111(4):1617–1622. doi: 10.1083/jcb.111.4.1617. [DOI] [PMC free article] [PubMed] [Google Scholar]
  8. 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]
  9. 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]
  10. Sheetz M. P., Baumrind N. L., Wayne D. B., Pearlman A. L. Concentration of membrane antigens by forward transport and trapping in neuronal growth cones. Cell. 1990 Apr 20;61(2):231–241. doi: 10.1016/0092-8674(90)90804-n. [DOI] [PubMed] [Google Scholar]
  11. Subramaniam S., Seul M., McConnell H. M. Lateral diffusion of specific antibodies bound to lipid monolayers on alkylated substrates. Proc Natl Acad Sci U S A. 1986 Mar;83(5):1169–1173. doi: 10.1073/pnas.83.5.1169. [DOI] [PMC free article] [PubMed] [Google Scholar]
  12. Wright L. L., Palmer A. G., 3rd, Thompson N. L. Inhomogeneous translational diffusion of monoclonal antibodies on phospholipid Langmuir-Blodgett films. Biophys J. 1988 Sep;54(3):463–470. doi: 10.1016/S0006-3495(88)82979-5. [DOI] [PMC free article] [PubMed] [Google Scholar]
  13. Wu E. S., Jacobson K., Szoka F., Portis A., Jr Lateral diffusion of a hydrophobic peptide, N-4-nitrobenz-2-oxa-1,3-diazole gramicidin S, in phospholipid multibilayers. Biochemistry. 1978 Dec 12;17(25):5543–5550. doi: 10.1021/bi00618a033. [DOI] [PubMed] [Google Scholar]
  14. 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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