Skip to main content
Biophysical Journal logoLink to Biophysical Journal
. 1998 Jun;74(6):2983–2995. doi: 10.1016/S0006-3495(98)78005-1

Phase transitions in films of lung surfactant at the air-water interface.

K Nag 1, J Perez-Gil 1, M L Ruano 1, L A Worthman 1, J Stewart 1, C Casals 1, K M Keough 1
PMCID: PMC1299639  PMID: 9635752

Abstract

Pulmonary surfactant maintains a putative surface-active film at the air-alveolar fluid interface and prevents lung collapse at low volumes. Porcine lung surfactant extracts (LSE) were studied in spread and adsorbed films at 23 +/- 1 degrees C using epifluorescence microscopy combined with surface balance techniques. By incorporating small amounts of fluorescent probe 1-palmitoyl-2-nitrobenzoxadiazole dodecanoyl phosphatidylcholine (NBD-PC) in LSE films the expanded (fluid) to condensed (gel-like) phase transition was studied under different compression rates and ionic conditions. Films spread from solvent and adsorbed from vesicles both showed condensed (probe-excluding) domains dispersed in a background of expanded (probe-including) phase, and the appearance of the films was similar at similar surface pressure. In quasistatically compressed LSE films the appearance of condensed domains occurred at a surface pressure (pi) of 13 mN/m. Such domains increased in size and amounts as pi was increased to 35 mN/m, and their amounts appeared to decrease to 4% upon further compression to 45 mN/m. Above pi of 45 mN/m the LSE films had the appearance of filamentous materials of finely divided dark and light regions, and such features persisted up to a pi near 68 mN/m. Some of the condensed domains had typical kidney bean shapes, and their distribution was similar to those seen previously in films of dipalmitoylphosphatidylcholine (DPPC), the major component of surfactant. Rapid cyclic compression and expansion of LSE films resulted in features that indicated a possible small (5%) loss of fluid components from such films or an increase in condensation efficiency over 10 cycles. Calcium (5 mM) in the subphase of LSE films altered the domain distribution, decreasing the size and increasing the number and total amount of condensed phase domains. Calcium also caused an increase in the value of pi at which the maximum amount of independent condensed phase domains were observed to 45 mN/m. It also induced formation of large amounts of novel, nearly circular domains containing probe above pi of 50 mN/m, these domains being different in appearance than any seen at lower pressures with calcium or higher pressures in the absence of calcium. Surfactant protein-A (SP-A) adsorbed from the subphase onto solvent-spread LSE films, and aggregated condensed domains in presence of calcium. This study indicates that spread or adsorbed lung surfactant films can undergo expanded to condensed, and possibly other, phase transitions at the air-water interface as lateral packing density increases. These phase transitions are affected by divalent cations and SP-A in the subphase, and possibly by loss of material from the surface upon cyclic compression and expansion.

Full Text

The Full Text of this article is available as a PDF (696.9 KB).

Selected References

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

  1. Amirkhanian J. D., Merritt T. A. The influence of pH on surface properties of lung surfactants. Lung. 1995;173(4):243–254. doi: 10.1007/BF00181876. [DOI] [PubMed] [Google Scholar]
  2. Amirkhanian J. D., Taeusch H. W. Reversible and irreversible inactivation of preformed pulmonary surfactant surface films by changes in subphase constituents. Biochim Biophys Acta. 1993 Jan 10;1165(3):321–326. doi: 10.1016/0005-2760(93)90143-w. [DOI] [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. BLIGH E. G., DYER W. J. A rapid method of total lipid extraction and purification. Can J Biochem Physiol. 1959 Aug;37(8):911–917. doi: 10.1139/o59-099. [DOI] [PubMed] [Google Scholar]
  5. Bachofen H., Schürch S., Urbinelli M., Weibel E. R. Relations among alveolar surface tension, surface area, volume, and recoil pressure. J Appl Physiol (1985) 1987 May;62(5):1878–1887. doi: 10.1152/jappl.1987.62.5.1878. [DOI] [PubMed] [Google Scholar]
  6. Bangham A. D. Lung surfactant: how it does and does not work. Lung. 1987;165(1):17–25. doi: 10.1007/BF02714417. [DOI] [PubMed] [Google Scholar]
  7. Benson B. J., Williams M. C., Sueishi K., Goerke J., Sargeant T. Role of calcium ions the structure and function of pulmonary surfactant. Biochim Biophys Acta. 1984 Mar 27;793(1):18–27. doi: 10.1016/0005-2760(84)90048-1. [DOI] [PubMed] [Google Scholar]
  8. Casals C., Miguel E., Perez-Gil J. Tryptophan fluorescence study on the interaction of pulmonary surfactant protein A with phospholipid vesicles. Biochem J. 1993 Dec 15;296(Pt 3):585–593. doi: 10.1042/bj2960585. [DOI] [PMC free article] [PubMed] [Google Scholar]
  9. Covey T. R., Huang E. C., Henion J. D. Structural characterization of protein tryptic peptides via liquid chromatography/mass spectrometry and collision-induced dissociation of their doubly charged molecular ions. Anal Chem. 1991 Jul 1;63(13):1193–1200. doi: 10.1021/ac00013a003. [DOI] [PubMed] [Google Scholar]
  10. Discher B. M., Maloney K. M., Schief W. R., Jr, Grainger D. W., Vogel V., Hall S. B. Lateral phase separation in interfacial films of pulmonary surfactant. Biophys J. 1996 Nov;71(5):2583–2590. doi: 10.1016/S0006-3495(96)79450-X. [DOI] [PMC free article] [PubMed] [Google Scholar]
  11. Dluhy R. A., Reilly K. E., Hunt R. D., Mitchell M. L., Mautone A. J., Mendelsohn R. Infrared spectroscopic investigations of pulmonary surfactant. Surface film transitions at the air-water interface and bulk phase thermotropism. Biophys J. 1989 Dec;56(6):1173–1181. doi: 10.1016/S0006-3495(89)82764-X. [DOI] [PMC free article] [PubMed] [Google Scholar]
  12. Efrati H., Hawgood S., Williams M. C., Hong K., Benson B. J. Divalent cation and hydrogen ion effects on the structure and surface activity of pulmonary surfactant. Biochemistry. 1987 Dec 1;26(24):7986–7993. doi: 10.1021/bi00398a066. [DOI] [PubMed] [Google Scholar]
  13. Flörsheimer M., Möhwald H. Development of equilibrium domain shapes in phospholipid monolayers. Chem Phys Lipids. 1989 Mar;49(4):231–241. doi: 10.1016/0009-3084(89)90071-6. [DOI] [PubMed] [Google Scholar]
  14. Gross N. J. Extracellular metabolism of pulmonary surfactant: the role of a new serine protease. Annu Rev Physiol. 1995;57:135–150. doi: 10.1146/annurev.ph.57.030195.001031. [DOI] [PubMed] [Google Scholar]
  15. Gulik A., Tchoreloff P., Proust J. A conformation transition of lung surfactant lipids probably involved in respiration. Biophys J. 1994 Sep;67(3):1107–1112. doi: 10.1016/S0006-3495(94)80576-4. [DOI] [PMC free article] [PubMed] [Google Scholar]
  16. Haddad I. Y., Holm B. A., Hlavaty L., Matalon S. Dependence of surfactant function on extracellular pH: mechanisms and modifications. J Appl Physiol (1985) 1994 Feb;76(2):657–662. doi: 10.1152/jappl.1994.76.2.657. [DOI] [PubMed] [Google Scholar]
  17. Hook G. E., Spalding J. W., Ortner M. J., Tombropoulos E. G., Chignell C. F. Investigation of phospholipids of the pulmonary extracellular lining by electron paramagnetic resonance. The effects of phosphatidylglycerol and unsaturated phosphatidylcholines on the fluidity of dipalmitoyl phosphatidylcholine. Biochem J. 1984 Oct 15;223(2):533–542. doi: 10.1042/bj2230533. [DOI] [PMC free article] [PubMed] [Google Scholar]
  18. 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]
  19. Keough K. M., Farrell E., Cox M., Harrell G., Taeusch H. W., Jr Physical, chemical, and physiological characteristics of isolates of pulmonary surfactant from adult rabbits. Can J Physiol Pharmacol. 1985 Sep;63(9):1043–1051. doi: 10.1139/y85-171. [DOI] [PubMed] [Google Scholar]
  20. Keough K. M., Kariel N. Differential scanning calorimetric studies of aqueous dispersions of phosphatidylcholines containing two polyenoic chains. Biochim Biophys Acta. 1987 Aug 7;902(1):11–18. doi: 10.1016/0005-2736(87)90130-1. [DOI] [PubMed] [Google Scholar]
  21. King R. J., Clements J. A. Surface active materials from dog lung. 3. Thermal analysis. Am J Physiol. 1972 Sep;223(3):727–733. doi: 10.1152/ajplegacy.1972.223.3.727. [DOI] [PubMed] [Google Scholar]
  22. King R. J., Phillips M. C., Horowitz P. M., Dang S. C. Interaction between the 35 kDa apolipoprotein of pulmonary surfactant and saturated phosphatidylcholines. Effects of temperature. Biochim Biophys Acta. 1986 Oct 24;879(1):1–13. doi: 10.1016/0005-2760(86)90259-6. [DOI] [PubMed] [Google Scholar]
  23. Kuroki Y., Akino T. Pulmonary surfactant protein A (SP-A) specifically binds dipalmitoylphosphatidylcholine. J Biol Chem. 1991 Feb 15;266(5):3068–3073. [PubMed] [Google Scholar]
  24. Lalchev Z. I., Todorov R. K., Christova Y. T., Wilde P. J., Mackie A. R., Clark D. C. Molecular mobility in the monolayers of foam films stabilized by porcine lung surfactant. Biophys J. 1996 Nov;71(5):2591–2601. doi: 10.1016/S0006-3495(96)79451-1. [DOI] [PMC free article] [PubMed] [Google Scholar]
  25. Mautone A. J., Reilly K. E., Mendelsohn R. Fourier transform infrared and differential scanning calorimetric studies of a surface-active material from rabbit lung. Biochim Biophys Acta. 1987 Jan 9;896(1):1–10. doi: 10.1016/0005-2736(87)90349-x. [DOI] [PubMed] [Google Scholar]
  26. Möhwald H. Phospholipid and phospholipid-protein monolayers at the air/water interface. Annu Rev Phys Chem. 1990;41:441–476. doi: 10.1146/annurev.pc.41.100190.002301. [DOI] [PubMed] [Google Scholar]
  27. Nag K., Boland C., Rich N., Keough K. M. Epifluorescence microscopic observation of monolayers of dipalmitoylphosphatidylcholine: dependence of domain size on compression rates. Biochim Biophys Acta. 1991 Sep 30;1068(2):157–160. doi: 10.1016/0005-2736(91)90204-l. [DOI] [PubMed] [Google Scholar]
  28. Nag K., Keough K. M. Epifluorescence microscopic studies of monolayers containing mixtures of dioleoyl- and dipalmitoylphosphatidylcholines. Biophys J. 1993 Sep;65(3):1019–1026. doi: 10.1016/S0006-3495(93)81155-X. [DOI] [PMC free article] [PubMed] [Google Scholar]
  29. Nag K., Perez-Gil J., Cruz A., Keough K. M. Fluorescently labeled pulmonary surfactant protein C in spread phospholipid monolayers. Biophys J. 1996 Jul;71(1):246–256. doi: 10.1016/S0006-3495(96)79221-4. [DOI] [PMC free article] [PubMed] [Google Scholar]
  30. Nag K., Perez-Gil J., Cruz A., Rich N. H., Keough K. M. Spontaneous formation of interfacial lipid-protein monolayers during adsorption from vesicles. Biophys J. 1996 Sep;71(3):1356–1363. doi: 10.1016/S0006-3495(96)79338-4. [DOI] [PMC free article] [PubMed] [Google Scholar]
  31. Oldfield E., Keough K. M., Chapman D. The study of hydrocarbon chain mobility in membrane systems using spin-label probes. FEBS Lett. 1972 Feb 15;20(3):344–346. doi: 10.1016/0014-5793(72)80103-0. [DOI] [PubMed] [Google Scholar]
  32. Otis D. R., Jr, Ingenito E. P., Kamm R. D., Johnson M. Dynamic surface tension of surfactant TA: experiments and theory. J Appl Physiol (1985) 1994 Dec;77(6):2681–2688. doi: 10.1152/jappl.1994.77.6.2681. [DOI] [PubMed] [Google Scholar]
  33. Pérez-Gil J., Nag K., Taneva S., Keough K. M. Pulmonary surfactant protein SP-C causes packing rearrangements of dipalmitoylphosphatidylcholine in spread monolayers. Biophys J. 1992 Jul;63(1):197–204. doi: 10.1016/S0006-3495(92)81582-5. [DOI] [PMC free article] [PubMed] [Google Scholar]
  34. Ruano M. L., Miguel E., Perez-Gil J., Casals C. Comparison of lipid aggregation and self-aggregation activities of pulmonary surfactant-associated protein A. Biochem J. 1996 Jan 15;313(Pt 2):683–689. doi: 10.1042/bj3130683. [DOI] [PMC free article] [PubMed] [Google Scholar]
  35. Ruano M. L., Nag K., Worthman L. A., Casals C., Pérez-Gil J., Keough K. M. Differential partitioning of pulmonary surfactant protein SP-A into regions of monolayers of dipalmitoylphosphatidylcholine and dipalmitoylphosphatidylcholine/dipalmitoylphosphatidylglycerol. Biophys J. 1998 Mar;74(3):1101–1109. doi: 10.1016/s0006-3495(98)77828-2. [DOI] [PMC free article] [PubMed] [Google Scholar]
  36. Scarpelli E. M., Mautone A. J. Surface biophysics of the surface monolayer theory is incompatible with regional lung function. Biophys J. 1994 Sep;67(3):1080–1089. doi: 10.1016/S0006-3495(94)80573-9. [DOI] [PMC free article] [PubMed] [Google Scholar]
  37. Schürch S., Bachofen H., Goerke J., Green F. Surface properties of rat pulmonary surfactant studied with the captive bubble method: adsorption, hysteresis, stability. Biochim Biophys Acta. 1992 Jan 10;1103(1):127–136. doi: 10.1016/0005-2736(92)90066-u. [DOI] [PubMed] [Google Scholar]
  38. Schürch S., Bachofen H., Weibel E. R. Alveolar surface tensions in excised rabbit lungs: effect of temperature. Respir Physiol. 1985 Oct;62(1):31–45. doi: 10.1016/0034-5687(85)90048-9. [DOI] [PubMed] [Google Scholar]
  39. Schürch S., Goerke J., Clements J. A. Direct determination of surface tension in the lung. Proc Natl Acad Sci U S A. 1976 Dec;73(12):4698–4702. doi: 10.1073/pnas.73.12.4698. [DOI] [PMC free article] [PubMed] [Google Scholar]
  40. Schürch S., Schürch D., Curstedt T., Robertson B. Surface activity of lipid extract surfactant in relation to film area compression and collapse. J Appl Physiol (1985) 1994 Aug;77(2):974–986. doi: 10.1152/jappl.1994.77.2.974. [DOI] [PubMed] [Google Scholar]
  41. Stine K. J. Investigations of monolayers by fluorescence microscopy. Microsc Res Tech. 1994 Apr 1;27(5):439–450. doi: 10.1002/jemt.1070270510. [DOI] [PubMed] [Google Scholar]
  42. Teubner J. K., Gibson R. A., McMurchie E. J. The influence of water on the phase transition of sheep lung surfactant. A possible mechanism for surfactant phase transitions in vivo. Biochim Biophys Acta. 1983 Mar 1;750(3):521–525. doi: 10.1016/0005-2760(83)90192-3. [DOI] [PubMed] [Google Scholar]
  43. Träuble H., Eibl H., Sawada H. Respiration--a critical phenomenon? Lipid phase transitions in the lung alveolar surfactant. Naturwissenschaften. 1974 Aug;61(8):344–354. doi: 10.1007/BF00600300. [DOI] [PubMed] [Google Scholar]
  44. Weis R. M. Fluorescence microscopy of phospholipid monolayer phase transitions. Chem Phys Lipids. 1991 Mar;57(2-3):227–239. doi: 10.1016/0009-3084(91)90078-p. [DOI] [PubMed] [Google Scholar]
  45. Williams M. C., Hawgood S., Hamilton R. L. Changes in lipid structure produced by surfactant proteins SP-A, SP-B, and SP-C. Am J Respir Cell Mol Biol. 1991 Jul;5(1):41–50. doi: 10.1165/ajrcmb/5.1.41. [DOI] [PubMed] [Google Scholar]
  46. Yu S. H., Possmayer F. Adsorption, compression and stability of surface films from natural, lipid extract and reconstituted pulmonary surfactants. Biochim Biophys Acta. 1993 Apr 23;1167(3):264–271. doi: 10.1016/0005-2760(93)90228-2. [DOI] [PubMed] [Google Scholar]

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

RESOURCES