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. 1981 Feb;33(2):211–223. doi: 10.1016/S0006-3495(81)84882-5

A biophysical study of protein-lipid interactions in membranes of Escherichia coli. Fluoromyristic acid as a probe.

M P Gent, P F Cottam, C Ho
PMCID: PMC1327421  PMID: 7013842

Abstract

Fluorine-19 nuclear magentic resonance spectroscopy and transport assays have been used to investigate and compare the membrane properties of unsaturated fatty acid auxotrophs of two strains of Escherichia coli, K1060B5 and ML 308-225-UFA-8. A fluorinated analog of myristic acid, 8, 8-difluoromyristic acid, can be incorporated into the membrane phospholipids by substitution for oleate in the growth medium. Growth for one generation on 8, 8-difluoromyristate results in a 20% content of fluorinated fatty acid in the membranes, changes in the protein to lipid ratio, and altered transport of methyl beta-D-thiogalactopyranoside. The differences in membrane composition and transport behavior seen in oleate supplemented E. coli K1060B5 relative to ML 308-225-UFA-8 are enhanced by the incorporation of 8, 8-difluoromyristate. The phase transition behavior becomes distinctly different and some differences in lipid organization persist above the transition temperature. Concomitantly, the rate and extent of concentration of methyl beta-D-thiogalactopyranoside are reduced two-fold more in E. coli K1060B5 compared to ML 308-225-UFA-8. Such behavior suggests that these fluorinated fatty acid supplemented strains of E. coli are useful to study subtle differences in protein-lipid interactions and their effects on the function of membrane-bound enzymes.

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Selected References

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

  1. Ames G. F. Resolution of bacterial proteins by polyacrylamide gel electrophoresis on slabs. Membrane, soluble, and periplasmic fractions. J Biol Chem. 1974 Jan 25;249(2):634–644. [PubMed] [Google Scholar]
  2. 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]
  3. Baldassare J. J., Breneckle G. M., Hoffman M., Silbert D. F. Modification of membrane lipid. Functional properties of membrane in relation to fatty acid structure. J Biol Chem. 1977 Dec 25;252(24):8797–8803. [PubMed] [Google Scholar]
  4. Baldassare J. J., Rhinehart K. B., Silbert D. F. Modification of membrane lipid: physical properties in relation to fatty acid structure. Biochemistry. 1976 Jul 13;15(14):2986–2994. doi: 10.1021/bi00659a008. [DOI] [PubMed] [Google Scholar]
  5. Bayer M. E., Dolack M., Houser E. Effects of lipid phase transition of the freeze-cleaved envelope of Escherichia coli. J Bacteriol. 1977 Mar;129(3):1563–1573. doi: 10.1128/jb.129.3.1563-1573.1977. [DOI] [PMC free article] [PubMed] [Google Scholar]
  6. Chapman D. Phase transitions and fluidity characteristics of lipids and cell membranes. Q Rev Biophys. 1975 May;8(2):185–235. doi: 10.1017/s0033583500001797. [DOI] [PubMed] [Google Scholar]
  7. Fox C. F., Carter J. R., Kennedy E. P. GENETIC CONTROL OF THE MEMBRANE PROTEIN COMPONENT OF THE LACTOSE TRANSPORT SYSTEM OF Escherichia coli. Proc Natl Acad Sci U S A. 1967 Mar;57(3):698–705. doi: 10.1073/pnas.57.3.698. [DOI] [PMC free article] [PubMed] [Google Scholar]
  8. Fried V. A. A novel mutant of the lac transport system of Escherichia coli. J Mol Biol. 1977 Aug 25;114(4):477–490. doi: 10.1016/0022-2836(77)90173-5. [DOI] [PubMed] [Google Scholar]
  9. Gent M. P., Cottam P. F., Ho C. Fluorine-19 nuclear magnetic resonance studies of Escherichia coli membranes. Proc Natl Acad Sci U S A. 1978 Feb;75(2):630–634. doi: 10.1073/pnas.75.2.630. [DOI] [PMC free article] [PubMed] [Google Scholar]
  10. Gent M. P., Ho C. Fluorine-19 nuclear magnetic resonance studies of lipid phase transitions in model and biological membranes. Biochemistry. 1978 Jul 25;17(15):3023–3038. doi: 10.1021/bi00608a014. [DOI] [PubMed] [Google Scholar]
  11. HORECKER B. L., THOMAS J., MONOD J. Galactose transport in Escherichia coli. I. General properties as studied in a galactokinaseless mutant. J Biol Chem. 1960 Jun;235:1580–1585. [PubMed] [Google Scholar]
  12. Hong J. S. An ecf mutation in Escherichia coli pleiotropically affecting energy coupling in active transport but not generation or maintenance of membrane potential. J Biol Chem. 1977 Dec 10;252(23):8582–8588. [PubMed] [Google Scholar]
  13. Hong J. S., Kaback H. R. Mutants of Salmonella typhimurium and Escherichia coli pleiotropically defective in active transport. Proc Natl Acad Sci U S A. 1972 Nov;69(11):3336–3340. doi: 10.1073/pnas.69.11.3336. [DOI] [PMC free article] [PubMed] [Google Scholar]
  14. Jones N. C., Osborn M. J. Interaction of Salmonella typhimurium with phospholipid vesicles. Incorporation of exogenous lipids into intact cells. J Biol Chem. 1977 Oct 25;252(20):7398–7404. [PubMed] [Google Scholar]
  15. Jones N. C., Osborn M. J. Translocation of phospholipids between the outer and inner membranes of Salmonella typhimurium. J Biol Chem. 1977 Oct 25;252(20):7405–7412. [PubMed] [Google Scholar]
  16. Jones T. H., Kennedy E. P. Characterization of the membrane protein component of the lactose transport system of Escherichia coli. J Biol Chem. 1969 Nov 10;244(21):5981–5987. [PubMed] [Google Scholar]
  17. Jost P. C., Griffith O. H., Capaldi R. A., Vanderkooi G. Evidence for boundary lipid in membranes. Proc Natl Acad Sci U S A. 1973 Feb;70(2):480–484. doi: 10.1073/pnas.70.2.480. [DOI] [PMC free article] [PubMed] [Google Scholar]
  18. Jost P. C., Nadakavukaren K. K., Griffith O. H. Phosphatidylcholine exchange between the boundary lipid and bilayer domains in cytochrome oxidase containing membranes. Biochemistry. 1977 Jul 12;16(14):3110–3114. doi: 10.1021/bi00633a011. [DOI] [PubMed] [Google Scholar]
  19. Kleemann W., McConnell H. M. Lateral phase separations in Escherichia coli membranes. Biochim Biophys Acta. 1974 Apr 29;345(2):220–230. doi: 10.1016/0005-2736(74)90260-0. [DOI] [PubMed] [Google Scholar]
  20. LENNOX E. S. Transduction of linked genetic characters of the host by bacteriophage P1. Virology. 1955 Jul;1(2):190–206. doi: 10.1016/0042-6822(55)90016-7. [DOI] [PubMed] [Google Scholar]
  21. LOWRY O. H., ROSEBROUGH N. J., FARR A. L., RANDALL R. J. Protein measurement with the Folin phenol reagent. J Biol Chem. 1951 Nov;193(1):265–275. [PubMed] [Google Scholar]
  22. Lancaster J. R., Jr, Hinkle P. C. Studies of the beta-galactoside transporter in inverted membrane vesicles of Escherichia coli. I. Symmetrical facilitated diffusion and proton gradient-coupled transport. J Biol Chem. 1977 Nov 10;252(21):7657–7661. [PubMed] [Google Scholar]
  23. Lancaster J. R., Jr, Hinkle P. C. Studies of the beta-galactoside transporter in inverted membrane vesicles of Escherichia coli. II. Symmetrical binding of a dansylgalactoside induced by an electrochemical proton gradient and by lactose efflux. J Biol Chem. 1977 Nov 10;252(21):7662–7666. [PubMed] [Google Scholar]
  24. Lee A. G. Lipid phase transitions and phase diagrams. I. Lipid phase transitions. Biochim Biophys Acta. 1977 Aug 9;472(2):237–281. doi: 10.1016/0304-4157(77)90018-1. [DOI] [PubMed] [Google Scholar]
  25. Letellier L., Moudden H., Shechter E. Lipid and protein segregation in Escherichia coli membrane: morphological and structural study of different cytoplasmic membrane fractions. Proc Natl Acad Sci U S A. 1977 Feb;74(2):452–456. doi: 10.1073/pnas.74.2.452. [DOI] [PMC free article] [PubMed] [Google Scholar]
  26. Linden C. D., Wright K. L., McConnell H. M., Fox C. F. Lateral phase separations in membrane lipids and the mechanism of sugar transport in Escherichia coli. Proc Natl Acad Sci U S A. 1973 Aug;70(8):2271–2275. doi: 10.1073/pnas.70.8.2271. [DOI] [PMC free article] [PubMed] [Google Scholar]
  27. Ohta T., Okuda S., Takahashi H. Relationship between phospholipid compositions and transport activities of amino acids in Escherichia coli membrane vesicles. Biochim Biophys Acta. 1977 Apr 1;466(1):44–56. doi: 10.1016/0005-2736(77)90207-3. [DOI] [PubMed] [Google Scholar]
  28. Overath P., Brenner M., Gulik-Krzywicki T., Shechter E., Letellier L. Lipid phase transitions in cytoplasmic and outer membranes of Escherichia coli. Biochim Biophys Acta. 1975 May 6;389(2):358–369. doi: 10.1016/0005-2736(75)90328-4. [DOI] [PubMed] [Google Scholar]
  29. Paton J. C., May B. K., Elliott W. H. Membrane phospholipid asymmetry in Bacillus amyloliquefaciens. J Bacteriol. 1978 Aug;135(2):393–401. doi: 10.1128/jb.135.2.393-401.1978. [DOI] [PMC free article] [PubMed] [Google Scholar]
  30. Petersen N. O., Chan S. I. More on the motional state of lipid bilayer membranes: interpretation of order parameters obtained from nuclear magnetic resonance experiments. Biochemistry. 1977 Jun 14;16(12):2657–2667. doi: 10.1021/bi00631a012. [DOI] [PubMed] [Google Scholar]
  31. Rotman B., Ganesan A. K., Guzman R. Transport systems for galactose and galactosides in Escherichia coli. II. Substrate and inducer specificities. J Mol Biol. 1968 Sep 14;36(2):247–260. doi: 10.1016/0022-2836(68)90379-3. [DOI] [PubMed] [Google Scholar]
  32. Sheetz M. P., Chan S. I. Effect of sonication on the structure of lecithin bilayers. Biochemistry. 1972 Nov 21;11(24):4573–4581. doi: 10.1021/bi00774a024. [DOI] [PubMed] [Google Scholar]
  33. 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]
  34. Silbert D. F., Ladenson R. C., Honegger J. L. The unsaturated fatty acid requirement in Escherichia coli. Temperature dependence and total replacement by branched-chain fatty acids. Biochim Biophys Acta. 1973 Jul 6;311(3):349–361. doi: 10.1016/0005-2736(73)90315-5. [DOI] [PubMed] [Google Scholar]
  35. 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]
  36. Smyth C. J., Siegel J., Salton M. R., Owen P. Immunochemical analysis of inner and outer membranes of Escherichia coli by crossed immunoelectrophoresis. J Bacteriol. 1978 Jan;133(1):306–319. doi: 10.1128/jb.133.1.306-319.1978. [DOI] [PMC free article] [PubMed] [Google Scholar]
  37. Sullivan K. H., Jain M. K., Koch A. L. Activation of the beta-galactoside transport system in Escherichia coli ML-308 by n-alkanols. Modification of lipid-protein interaction by a change in bilayer fluidity. Biochim Biophys Acta. 1974 Jun 13;352(2):287–297. doi: 10.1016/0005-2736(74)90220-x. [DOI] [PubMed] [Google Scholar]
  38. Teather R. M., Hamelin O., Schwarz H., Overath P. Functional symmetry of the beta-galactoside carrier in Escherichia coli. Biochim Biophys Acta. 1977 Jun 16;467(3):386–395. doi: 10.1016/0005-2736(77)90316-9. [DOI] [PubMed] [Google Scholar]
  39. Therisod H., Letellier L., Weil R., Shechter E. Functional lac carrier proteins in cytoplasmic membrane vesicles isolated from Escherichia coli. 1. Temperature dependence of dansyl galactoside binding and beta-galactoside transport. Biochemistry. 1977 Aug 23;16(17):3772–3776. doi: 10.1021/bi00636a007. [DOI] [PubMed] [Google Scholar]
  40. Vanderwinkel E., De Vlieghere M., Fontaine M., Charles D., Denamur F., Vandevoorde D., De Kegel D. Septation deficiency and phosphilipid perturbation in Escherichia coli genetically constitutive for the beta oxidation pathway. J Bacteriol. 1976 Sep;127(3):1389–1399. doi: 10.1128/jb.127.3.1389-1399.1976. [DOI] [PMC free article] [PubMed] [Google Scholar]
  41. West I. C., Wilson T. H. Galactoside transport dissociated from proton movement in mutants of Escherichia coli. Biochem Biophys Res Commun. 1973 Jan 23;50(2):551–558. doi: 10.1016/0006-291x(73)90875-9. [DOI] [PubMed] [Google Scholar]
  42. Wilson T. H., Kusch M. A mutant of Escherichia coli K 12 energy-uncoupled for lactose transport. Biochim Biophys Acta. 1972 Mar 17;255(3):786–797. doi: 10.1016/0005-2736(72)90391-4. [DOI] [PubMed] [Google Scholar]
  43. Winkler H. H., Wilson T. H. The role of energy coupling in the transport of beta-galactosides by Escherichia coli. J Biol Chem. 1966 May 25;241(10):2200–2211. [PubMed] [Google Scholar]
  44. Yang S. F., Freer S., Benson A. A. Transphosphatidylation by phospholipase D. J Biol Chem. 1967 Feb 10;242(3):477–484. [PubMed] [Google Scholar]
  45. Yariv J., Kalb A. J., Katchalski E., Goldman R., Thomas E. W. Two locations of the lac permease sulphydryl in the membrane of E. coli. FEBS Lett. 1969 Nov 12;5(3):173–176. doi: 10.1016/0014-5793(69)80324-8. [DOI] [PubMed] [Google Scholar]
  46. van Heerikhuizen H., Kwak E., van Bruggen E. F., Witholt B. Characterization of a low density cytoplasmic membrane subfraction isolated from Escherichia coli. Biochim Biophys Acta. 1975 Dec 1;413(2):177–191. doi: 10.1016/0005-2736(75)90102-9. [DOI] [PubMed] [Google Scholar]

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