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. 2002 Sep;83(3):1511–1524. doi: 10.1016/S0006-3495(02)73921-0

Physical and photophysical characterization of a BODIPY phosphatidylcholine as a membrane probe.

Mohammed Dahim 1, Nancy K Mizuno 1, Xin-Min Li 1, William E Momsen 1, Maureen M Momsen 1, Howard L Brockman 1
PMCID: PMC1302249  PMID: 12202376

Abstract

Lipids containing the dimethyl BODIPY fluorophore are used in cell biology because their fluorescence properties change with fluorophore concentration (C.-S. Chen, O. C. Martin, and R. E. Pagano. 1997. Biophys J. 72:37-50). The miscibility and steady-state fluorescence behavior of one such lipid, 1-palmitoyl-2-(4,4-difluoro-5,7-dimethyl-4-bora-3a,4a-diaza-s-indacene-3-pentanoyl)-sn-glycero-3-phosphocholine (PBPC), have been characterized in mixtures with 1-stearoyl-2-oleoyl-sn-glycero-3-phosphocholine (SOPC). PBPC packs similarly to phosphatidylcholines having a cis-unsaturated acyl chain and mixes nearly ideally with SOPC, apparently without fluorophore-fluorophore aggregation. Increasing PBPC mole fraction from 0.0 to 1.0 in SOPC membranes changes the emission characteristics of the probe in a continuous manner. Analysis of these changes shows that emission from the excited dimethyl BODIPY monomer self quenches with a critical radius of 25.9 A. Fluorophores sufficiently close (< or =13.7 A) at the time of excitation can form an excited dimer, emission from which depends strongly on total lipid packing density. Overall, the data show that PBPC is a reasonable physical substitute for other phosphatidylcholines in fluid membranes. Knowledge of PBPC fluorescence in lipid monolayers has been exploited to determine the two-dimensional concentration of SOPC in unilamellar, bilayer 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. Ali S., Smaby J. M., Brockman H. L., Brown R. E. Cholesterol's interfacial interactions with galactosylceramides. Biochemistry. 1994 Mar 15;33(10):2900–2906. doi: 10.1021/bi00176a020. [DOI] [PMC free article] [PubMed] [Google Scholar]
  2. Ali S., Smaby J. M., Momsen M. M., Brockman H. L., Brown R. E. Acyl chain-length asymmetry alters the interfacial elastic interactions of phosphatidylcholines. Biophys J. 1998 Jan;74(1):338–348. doi: 10.1016/S0006-3495(98)77791-4. [DOI] [PMC free article] [PubMed] [Google Scholar]
  3. BARTLETT G. R. Phosphorus assay in column chromatography. J Biol Chem. 1959 Mar;234(3):466–468. [PubMed] [Google Scholar]
  4. BROCKMAN H. Dipole potential of lipid membranes. Chem Phys Lipids. 1994 Sep 6;73(1-2):57–79. doi: 10.1016/0009-3084(94)90174-0. [DOI] [PubMed] [Google Scholar]
  5. Bergström F., Hägglöf P., Karolin J., Ny T., Johansson L. B. The use of site-directed fluorophore labeling and donor-donor energy migration to investigate solution structure and dynamics in proteins. Proc Natl Acad Sci U S A. 1999 Oct 26;96(22):12477–12481. doi: 10.1073/pnas.96.22.12477. [DOI] [PMC free article] [PubMed] [Google Scholar]
  6. Caruso F., Grieser F., Thistlethwaite P. J., Almgren M. Two-dimensional diffusion of amphiphiles in phospholipid monolayers at the air-water interface. Biophys J. 1993 Dec;65(6):2493–2503. doi: 10.1016/S0006-3495(93)81301-8. [DOI] [PMC free article] [PubMed] [Google Scholar]
  7. Chen C. S., Martin O. C., Pagano R. E. Changes in the spectral properties of a plasma membrane lipid analog during the first seconds of endocytosis in living cells. Biophys J. 1997 Jan;72(1):37–50. doi: 10.1016/S0006-3495(97)78645-4. [DOI] [PMC free article] [PubMed] [Google Scholar]
  8. Chen C. S., Patterson M. C., Wheatley C. L., O'Brien J. F., Pagano R. E. Broad screening test for sphingolipid-storage diseases. Lancet. 1999 Sep 11;354(9182):901–905. doi: 10.1016/S0140-6736(98)10034-X. [DOI] [PubMed] [Google Scholar]
  9. Costigan S. C., Booth P. J., Templer R. H. Estimations of lipid bilayer geometry in fluid lamellar phases. Biochim Biophys Acta. 2000 Sep 29;1468(1-2):41–54. doi: 10.1016/s0005-2736(00)00220-0. [DOI] [PubMed] [Google Scholar]
  10. Farber S. A., Pack M., Ho S. Y., Johnson I. D., Wagner D. S., Dosch R., Mullins M. C., Hendrickson H. S., Hendrickson E. K., Halpern M. E. Genetic analysis of digestive physiology using fluorescent phospholipid reporters. Science. 2001 May 18;292(5520):1385–1388. doi: 10.1126/science.1060418. [DOI] [PubMed] [Google Scholar]
  11. Frijlink H. W., Eissens A. C., Hefting N. R., Poelstra K., Lerk C. F., Meijer D. K. The effect of parenterally administered cyclodextrins on cholesterol levels in the rat. Pharm Res. 1991 Jan;8(1):9–16. doi: 10.1023/a:1015861719134. [DOI] [PubMed] [Google Scholar]
  12. Galla H. J., Hartmann W., Theilen U., Sackmann E. On two-dimensional passive random walk in lipid bilayers and fluid pathways in biomembranes. J Membr Biol. 1979 Jul 31;48(3):215–236. doi: 10.1007/BF01872892. [DOI] [PubMed] [Google Scholar]
  13. Galla H. J., Sackmann E. Lateral diffusion in the hydrophobic region of membranes: use of pyrene excimers as optical probes. Biochim Biophys Acta. 1974 Feb 26;339(1):103–115. doi: 10.1016/0005-2736(74)90336-8. [DOI] [PubMed] [Google Scholar]
  14. Hwang J., Tamm L. K., Böhm, Ramalingam T. S., Betzig E., Edidin M. Nanoscale complexity of phospholipid monolayers investigated by near-field scanning optical microscopy. Science. 1995 Oct 27;270(5236):610–614. doi: 10.1126/science.270.5236.610. [DOI] [PubMed] [Google Scholar]
  15. Johnson I. D., Kang H. C., Haugland R. P. Fluorescent membrane probes incorporating dipyrrometheneboron difluoride fluorophores. Anal Biochem. 1991 Nov 1;198(2):228–237. doi: 10.1016/0003-2697(91)90418-s. [DOI] [PubMed] [Google Scholar]
  16. Kaiser R. D., London E. Determination of the depth of BODIPY probes in model membranes by parallax analysis of fluorescence quenching. Biochim Biophys Acta. 1998 Oct 15;1375(1-2):13–22. doi: 10.1016/s0005-2736(98)00127-8. [DOI] [PubMed] [Google Scholar]
  17. Kasurinen J. A novel fluorescent fatty acid, 5-methyl-BDY-3-dodecanoic acid, is a potential probe in lipid transport studies by incorporating selectively to lipid classes of BHK cells. Biochem Biophys Res Commun. 1992 Sep 30;187(3):1594–1601. doi: 10.1016/0006-291x(92)90485-4. [DOI] [PubMed] [Google Scholar]
  18. Koenig B. W., Strey H. H., Gawrisch K. Membrane lateral compressibility determined by NMR and x-ray diffraction: effect of acyl chain polyunsaturation. Biophys J. 1997 Oct;73(4):1954–1966. doi: 10.1016/S0006-3495(97)78226-2. [DOI] [PMC free article] [PubMed] [Google Scholar]
  19. Ladokhin A. S., Jayasinghe S., White S. H. How to measure and analyze tryptophan fluorescence in membranes properly, and why bother? Anal Biochem. 2000 Oct 15;285(2):235–245. doi: 10.1006/abio.2000.4773. [DOI] [PubMed] [Google Scholar]
  20. Lentz B. R., Burgess S. W. A dimerization model for the concentration dependent photophysical properties of diphenylhexatriene and its phospholipid derivatives. DPHpPC and DPHpPA. Biophys J. 1989 Oct;56(4):723–733. doi: 10.1016/S0006-3495(89)82720-1. [DOI] [PMC free article] [PubMed] [Google Scholar]
  21. Li X. M., Momsen M. M., Smaby J. M., Brockman H. L., Brown R. E. Cholesterol decreases the interfacial elasticity and detergent solubility of sphingomyelins. Biochemistry. 2001 May 22;40(20):5954–5963. doi: 10.1021/bi002791n. [DOI] [PMC free article] [PubMed] [Google Scholar]
  22. MacDonald R. C., MacDonald R. I., Menco B. P., Takeshita K., Subbarao N. K., Hu L. R. Small-volume extrusion apparatus for preparation of large, unilamellar vesicles. Biochim Biophys Acta. 1991 Jan 30;1061(2):297–303. doi: 10.1016/0005-2736(91)90295-j. [DOI] [PubMed] [Google Scholar]
  23. Marsh D. Lateral pressure in membranes. Biochim Biophys Acta. 1996 Oct 29;1286(3):183–223. doi: 10.1016/s0304-4157(96)00009-3. [DOI] [PubMed] [Google Scholar]
  24. Momsen M. M., Dahim M., Brockman H. L. Lateral packing of the pancreatic lipase cofactor, colipase, with phosphatidylcholine and substrates. Biochemistry. 1997 Aug 19;36(33):10073–10081. doi: 10.1021/bi9703857. [DOI] [PubMed] [Google Scholar]
  25. Pagano R. E., Martin O. C., Kang H. C., Haugland R. P. A novel fluorescent ceramide analogue for studying membrane traffic in animal cells: accumulation at the Golgi apparatus results in altered spectral properties of the sphingolipid precursor. J Cell Biol. 1991 Jun;113(6):1267–1279. doi: 10.1083/jcb.113.6.1267. [DOI] [PMC free article] [PubMed] [Google Scholar]
  26. Pagano R. E., Watanabe R., Wheatley C., Chen C. S. Use of N-[5-(5,7-dimethyl boron dipyrromethene difluoride-sphingomyelin to study membrane traffic along the endocytic pathway. Chem Phys Lipids. 1999 Nov;102(1-2):55–63. doi: 10.1016/s0009-3084(99)00075-4. [DOI] [PubMed] [Google Scholar]
  27. Peters R., Beck K. Translational diffusion in phospholipid monolayers measured by fluorescence microphotolysis. Proc Natl Acad Sci U S A. 1983 Dec;80(23):7183–7187. doi: 10.1073/pnas.80.23.7183. [DOI] [PMC free article] [PubMed] [Google Scholar]
  28. Rodriguez F., Tocanne J. F., Lopez A. Self-association processes involving anthracene labeled phosphatidylcholines in model membrane. Biophys Chem. 1995 Feb;53(3):169–180. doi: 10.1016/0301-4622(94)00109-w. [DOI] [PubMed] [Google Scholar]
  29. Smaby J. M., Brockman H. L. Surface dipole moments of lipids at the argon-water interface. Similarities among glycerol-ester-based lipids. Biophys J. 1990 Jul;58(1):195–204. doi: 10.1016/S0006-3495(90)82365-1. [DOI] [PMC free article] [PubMed] [Google Scholar]
  30. Stryer L., Haugland R. P. Energy transfer: a spectroscopic ruler. Proc Natl Acad Sci U S A. 1967 Aug;58(2):719–726. doi: 10.1073/pnas.58.2.719. [DOI] [PMC free article] [PubMed] [Google Scholar]
  31. Thuren T., Virtanen J. A., Kinnunen P. K. Estimation of the equilibrium lateral pressure in 1-palmitoyl-2-[6(pyren-1-yl)]hexanoyl-glycerophospholipid liposomes. Chem Phys Lipids. 1986 Oct-Nov;41(3-4):329–334. doi: 10.1016/0009-3084(86)90030-7. [DOI] [PubMed] [Google Scholar]

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