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. 1978 Jun;75(6):2759–2763. doi: 10.1073/pnas.75.6.2759

Determination of molecular motion in membranes using periodic pattern photobleaching.

B A Smith, H M McConnell
PMCID: PMC392643  PMID: 275845

Abstract

The lateral diffusion of a fluorescent phospholipid probe in oriented multibilayers of dimyristoylphosphatidylcholine has been measured by observing the redistribution of fluorescence after photobleaching of the membranes in a periodic pattern of parallel stripes. The diffusion constant D of the fluorescent lipid was found to vary between 1.5 X 10(-11) cm2/sec at 9.6 degrees and 2.0 X 10(-10) at 22.5 degrees in the monoclinic phase. Preliminary studies of dipalmitoylphosphatidylcholine liposomes in the Lbeta' and Pbeta' phases yielded diffusion constants of the order of 10(-11) cm2/sec. These data are relevant to earlier discussions of the rate of complement activation by hapten-sensitized liposomal membranes [Brûlet, P. and McConnell, H. M. (1976) Proc. Natl. Acad. Sci. USA 73,2977--2981; Parce, J. W., Henry, N. and McConnell, H.M. (1978) Proc. Natl. Acad. Sci. USA 75, 1515--1518)]. We have also used this method to study the motion of fluorescent antibodies bound to murine EL-4 tumor cells. Pattern photobleaching techniques have the advantages that cellular or liposomal translation has no major adverse effect on the measurements, that certain nondiffusive motions can be detected and characterized, and that diffusive or other motions can be recorded photographically.

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Selected References

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

  1. Badley R. A., Martin W. G., Schneider H. Dynamic behavior of fluorescent probes in lipid bilayer model membranes. Biochemistry. 1973 Jan 16;12(2):268–275. doi: 10.1021/bi00726a015. [DOI] [PubMed] [Google Scholar]
  2. Baumann G., Mueller P. A molecular model of membrane excitability. J Supramol Struct. 1974;2(5-6):538–557. doi: 10.1002/jss.400020504. [DOI] [PubMed] [Google Scholar]
  3. Brûlet P., McConnell H. M. Lateral hapten mobility and immunochemistry of model membranes. Proc Natl Acad Sci U S A. 1976 Sep;73(9):2977–2981. doi: 10.1073/pnas.73.9.2977. [DOI] [PMC free article] [PubMed] [Google Scholar]
  4. Brûlet P., McConnell H. M. Magnetic resonance spectra of membranes. Proc Natl Acad Sci U S A. 1975 Apr;72(4):1451–1455. doi: 10.1073/pnas.72.4.1451. [DOI] [PMC free article] [PubMed] [Google Scholar]
  5. Brûlet P., McConnell H. M. Structural and dynamical aspects of membrane immunochemistry using model membranes. Biochemistry. 1977 Mar 22;16(6):1209–1217. doi: 10.1021/bi00625a028. [DOI] [PubMed] [Google Scholar]
  6. Devaux P., McConnell H. M. Lateral diffusion in spin-labeled phosphatidylcholine multilayers. J Am Chem Soc. 1972 Jun 28;94(13):4475–4481. doi: 10.1021/ja00768a600. [DOI] [PubMed] [Google Scholar]
  7. Edelman G. M. Surface modulation in cell recognition and cell growth. Science. 1976 Apr 16;192(4236):218–226. doi: 10.1126/science.769162. [DOI] [PubMed] [Google Scholar]
  8. Fahey P. F., Koppel D. E., Barak L. S., Wolf D. E., Elson E. L., Webb W. W. Lateral diffusion in planar lipid bilayers. Science. 1977 Jan 21;195(4275):305–306. doi: 10.1126/science.831279. [DOI] [PubMed] [Google Scholar]
  9. Grant C. W., McConnell H. M. Glycophorin in lipid bilayers. Proc Natl Acad Sci U S A. 1974 Dec;71(12):4653–4657. doi: 10.1073/pnas.71.12.4653. [DOI] [PMC free article] [PubMed] [Google Scholar]
  10. Grant C. W., Wu S. H., McConnell H. M. Lateral phase separations in binary lipid mixtures: correlation between spin label and freeze-fracture electron microscopic studies. Biochim Biophys Acta. 1974 Sep 6;363(2):151–158. doi: 10.1016/0005-2736(74)90055-8. [DOI] [PubMed] [Google Scholar]
  11. Janiak M. J., Small D. M., Shipley G. G. Nature of the Thermal pretransition of synthetic phospholipids: dimyristolyl- and dipalmitoyllecithin. Biochemistry. 1976 Oct 19;15(21):4575–4580. doi: 10.1021/bi00666a005. [DOI] [PubMed] [Google Scholar]
  12. Kornberg R. D., McConnell H. M. Lateral diffusion of phospholipids in a vesicle membrane. Proc Natl Acad Sci U S A. 1971 Oct;68(10):2564–2568. doi: 10.1073/pnas.68.10.2564. [DOI] [PMC free article] [PubMed] [Google Scholar]
  13. Luna E. J., McConnell H. M. The intermediate monoclinic phase of phosphatidylcholines. Biochim Biophys Acta. 1977 May 2;466(3):381–392. doi: 10.1016/0005-2736(77)90331-5. [DOI] [PubMed] [Google Scholar]
  14. Mabrey S., Sturtevant J. M. Investigation of phase transitions of lipids and lipid mixtures by sensitivity differential scanning calorimetry. Proc Natl Acad Sci U S A. 1976 Nov;73(11):3862–3866. doi: 10.1073/pnas.73.11.3862. [DOI] [PMC free article] [PubMed] [Google Scholar]
  15. Mendoza G., Metzger H. Distribution and valency of receptor for IgE on rodent mast cells and related tumour cells. Nature. 1976 Dec 9;264(5586):548–550. doi: 10.1038/264548a0. [DOI] [PubMed] [Google Scholar]
  16. Nicolson G. L. Transmembrane control of the receptors on normal and tumor cells. I. Cytoplasmic influence over surface components. Biochim Biophys Acta. 1976 Apr 13;457(1):57–108. doi: 10.1016/0304-4157(76)90014-9. [DOI] [PubMed] [Google Scholar]
  17. Parce J. W., Henry N., McConnell H. M. Specific antibody-dependent binding of complement component C1q to hapten-sensitized lipid vesicles. Proc Natl Acad Sci U S A. 1978 Mar;75(3):1515–1518. doi: 10.1073/pnas.75.3.1515. [DOI] [PMC free article] [PubMed] [Google Scholar]
  18. 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]
  19. Segal D. M., Taurog J. D., Metzger H. Dimeric immunoglobulin E serves as a unit signal for mast cell degranulation. Proc Natl Acad Sci U S A. 1977 Jul;74(7):2993–2997. doi: 10.1073/pnas.74.7.2993. [DOI] [PMC free article] [PubMed] [Google Scholar]
  20. Shimshick E. J., McConnell H. M. Lateral phase separation in phospholipid membranes. Biochemistry. 1973 Jun 5;12(12):2351–2360. doi: 10.1021/bi00736a026. [DOI] [PubMed] [Google Scholar]
  21. Träuble H., Sackmann E. Studies of the crystalline-liquid crystalline phase transition of lipid model membranes. 3. Structure of a steroid-lecithin system below and above the lipid-phase transition. J Am Chem Soc. 1972 Jun 28;94(13):4499–4510. doi: 10.1021/ja00768a015. [DOI] [PubMed] [Google Scholar]

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