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. 1979 Dec;76(12):6245–6249. doi: 10.1073/pnas.76.12.6245

Energetics of rapid transmembrane movement and of compositional asymmetry of phosphatidylethanolamine in membranes of Bacillus megaterium.

K E Langley, E P Kennedy
PMCID: PMC411840  PMID: 118463

Abstract

The energy requirements for the rapid transmembrane movement of phosphatidylethanolamine in membranes of Bacillus megaterium KM have been investigated by means of pulse label experiments. The transmembrane movement continues at a high rate in cells blocked in the production of metabolic energy by treatment with a combination of inhibitors. The movement is shown to be completely independent of the synthesis of lipid and of protein and, more generally, independent of sources of metabolic energy. The rate constant ki, defined as the fraction of the internal phosphatidylethanolamine that exchanges with the external layer of the membrane per unit time, has been found to have a value of about 0.1 per min. The compositional asymmetry of phosphatidylethanolamine in membranes of B. megaterium persisted, and indeed was somewhat enhance, in energy-poisoned cells under conditions in which rapid mixing of inner and outer layers was taking place. Therefore, the compositional asymmetry is not maintained by kinetic barriers to transbilayer exchange or by expenditure of metabolic energy. It must be an equilibrium condition, and presumably reflects the differential binding of phospholipids by proteins and other ligands on the two sides of the membrane.

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Selected References

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  1. Bishop D. G., Bevers E. M., van Meer G., Op den Kamp J. A., van Deenen L. L. A monolayer study of the reaction of trinitrobenzene sulphonic acid with amino phospholipids. Biochim Biophys Acta. 1979 Feb 20;551(1):122–128. doi: 10.1016/0005-2736(79)90358-4. [DOI] [PubMed] [Google Scholar]
  2. Blobel G., Dobberstein B. Transfer of proteins across membranes. I. Presence of proteolytically processed and unprocessed nascent immunoglobulin light chains on membrane-bound ribosomes of murine myeloma. J Cell Biol. 1975 Dec;67(3):835–851. doi: 10.1083/jcb.67.3.835. [DOI] [PMC free article] [PubMed] [Google Scholar]
  3. Blobel G., Dobberstein B. Transfer of proteins across membranes. II. Reconstitution of functional rough microsomes from heterologous components. J Cell Biol. 1975 Dec;67(3):852–862. doi: 10.1083/jcb.67.3.852. [DOI] [PMC free article] [PubMed] [Google Scholar]
  4. Bloj B., Zilversmit D. B. Asymmetry and transposition rates of phosphatidylcholine in rat erythrocyte ghosts. Biochemistry. 1976 Mar 23;15(6):1277–1283. doi: 10.1021/bi00651a017. [DOI] [PubMed] [Google Scholar]
  5. Both G. W., McInnes J. L., Hanlon J. E., May B. K., Elliott W. H. Evidence for an accumulation of messenger RNA specific for extracellular protease and its relevance to the mechanism of enzyme secretion in bacteria. J Mol Biol. 1972 Jun 20;67(2):199–217. doi: 10.1016/0022-2836(72)90236-7. [DOI] [PubMed] [Google Scholar]
  6. Cronan J. E., Jr, Birge C. H., Vagelos P. R. Evidence for two genes specifically involved in unsaturated fatty acid biosynthesis in Escherichia coli. J Bacteriol. 1969 Nov;100(2):601–604. doi: 10.1128/jb.100.2.601-604.1969. [DOI] [PMC free article] [PubMed] [Google Scholar]
  7. De Kruijff B., Van Zoelen E. J. Effect of the phase transition on the transbilayer movement of dimyristoyl phosphatidylcholine in unilamellar vesicles. Biochim Biophys Acta. 1978 Jul 20;511(1):105–115. doi: 10.1016/0005-2736(78)90068-8. [DOI] [PubMed] [Google Scholar]
  8. 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]
  9. Grant C. W., McConnell H. M. Fusion of phospholipid vesicles with viable Acholeplasma laidlawii. Proc Natl Acad Sci U S A. 1973 Apr;70(4):1238–1240. doi: 10.1073/pnas.70.4.1238. [DOI] [PMC free article] [PubMed] [Google Scholar]
  10. Inouye H., Beckwith J. Synthesis and processing of an Escherichia coli alkaline phosphatase precursor in vitro. Proc Natl Acad Sci U S A. 1977 Apr;74(4):1440–1444. doi: 10.1073/pnas.74.4.1440. [DOI] [PMC free article] [PubMed] [Google Scholar]
  11. McNamee M. G., McConnell H. M. Transmembrane potentials and phospholipid flip-flop in excitable membrane vesicles. Biochemistry. 1973 Jul 31;12(16):2951–2958. doi: 10.1021/bi00740a001. [DOI] [PubMed] [Google Scholar]
  12. Renooij W., Van Golde L. M., Zwaal R. F., Van Deenen L. L. Topological asymmetry of phospholipid metabolism in rat erythrocyte membranes. Evidence for flip-flop of lecithin. Eur J Biochem. 1976 Jan 2;61(1):53–58. doi: 10.1111/j.1432-1033.1976.tb09996.x. [DOI] [PubMed] [Google Scholar]
  13. Roseman M., Litman B. J., Thompson T. E. Transbilayer exchange of phosphatidylethanolamine for phosphatidylcholine and N-acetimidoylphosphatidylethanolamine in single-walled bilayer vesicles. Biochemistry. 1975 Nov 4;14(22):4826–4830. doi: 10.1021/bi00693a008. [DOI] [PubMed] [Google Scholar]
  14. Rothman J. E., Kennedy E. P. Asymmetrical distribution of phospholipids in the membrane of Bacillus megaterium. J Mol Biol. 1977 Mar 5;110(3):603–618. doi: 10.1016/s0022-2836(77)80114-9. [DOI] [PubMed] [Google Scholar]
  15. Rothman J. E., Kennedy E. P. Rapid transmembrane movement of newly synthesized phospholipids during membrane assembly. Proc Natl Acad Sci U S A. 1977 May;74(5):1821–1825. doi: 10.1073/pnas.74.5.1821. [DOI] [PMC free article] [PubMed] [Google Scholar]
  16. Rothman J. E., Tsai D. K., Dawidowicz E. A., Lenard J. Transbilayer phospholipid asymmetry and its maintenance in the membrane of influenza virus. Biochemistry. 1976 Jun 1;15(11):2361–2370. doi: 10.1021/bi00656a018. [DOI] [PubMed] [Google Scholar]
  17. Rousselet A., Colbeau A., Vignais P. M., Devaux P. F. Study of the transverse diffusion of spin-labeled phospholipids in biological membranes. II. Inner mitochondrial membrane of rat liver: use of phosphatidylcholine exchange protein. Biochim Biophys Acta. 1976 Mar 19;426(3):372–384. doi: 10.1016/0005-2736(76)90383-7. [DOI] [PubMed] [Google Scholar]
  18. Rousselet A., Guthmann C., Matricon J., Bienvenue A., Devaux P. F. Study of the transverse diffusion of spin labeled phospholipids in biological membranes. I. Human red bloods cells. Biochim Biophys Acta. 1976 Mar 19;426(3):357–371. doi: 10.1016/0005-2736(76)90382-5. [DOI] [PubMed] [Google Scholar]
  19. Sackmann E., Träuble H., Galla H. J., Overath P. Lateral diffusion, protein mobility, and phase transitions in Escherichia coli membranes. A spin label study. Biochemistry. 1973 Dec 18;12(26):5360–5369. doi: 10.1021/bi00750a020. [DOI] [PubMed] [Google Scholar]
  20. Sandra A., Pagano R. E. Phospholipid asymmetry in LM cell plasma membrane derivatives: polar head group and acyl chain distributions. Biochemistry. 1978 Jan 24;17(2):332–338. doi: 10.1021/bi00595a022. [DOI] [PubMed] [Google Scholar]
  21. Shaw J. M., Moore N. F., Patzer E. J., Correa-Freire M. C., Wagner R. R., Thompsom T. E. Compositional asymmetry and transmembrane movement of phosphatidylcholine in vesicular stomatitis virus membranes. Biochemistry. 1979 Feb 6;18(3):538–543. doi: 10.1021/bi00570a024. [DOI] [PubMed] [Google Scholar]
  22. 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]
  23. Smith W. P., Tai P. C., Davis B. D. Interaction of secreted nascent chains with surrounding membrane in Bacillus subtilis. Proc Natl Acad Sci U S A. 1978 Dec;75(12):5922–5925. doi: 10.1073/pnas.75.12.5922. [DOI] [PMC free article] [PubMed] [Google Scholar]
  24. Stanley P. E., Williams S. G. Use of the liquid scintillation spectrometer for determining adenosine triphosphate by the luciferase enzyme. Anal Biochem. 1969 Jun;29(3):381–392. doi: 10.1016/0003-2697(69)90323-6. [DOI] [PubMed] [Google Scholar]
  25. Steck T. L. The organization of proteins in the human red blood cell membrane. A review. J Cell Biol. 1974 Jul;62(1):1–19. doi: 10.1083/jcb.62.1.1. [DOI] [PMC free article] [PubMed] [Google Scholar]
  26. Wirtz K. W. Transfer of phospholipids between membranes. Biochim Biophys Acta. 1974 Sep 16;344(2):95–117. doi: 10.1016/0304-4157(74)90001-x. [DOI] [PubMed] [Google Scholar]
  27. Zilversmit D. B., Hughes M. E. Extensive exchange of rat liver microsomal phospholipids. Biochim Biophys Acta. 1977 Aug 15;469(1):99–110. doi: 10.1016/0005-2736(77)90329-7. [DOI] [PubMed] [Google Scholar]
  28. Zilversmit D. B. Phospholipid-exchange proteins as membrane probes. Ann N Y Acad Sci. 1978;308:149–163. doi: 10.1111/j.1749-6632.1978.tb22020.x. [DOI] [PubMed] [Google Scholar]
  29. van den Besselaar A. M., de Druijff B., van den Bosch H., van Deenen L. L. Phosphatidylcholine mobility in liver microsomal membranes. Biochim Biophys Acta. 1978 Jul 4;510(2):242–255. doi: 10.1016/0005-2736(78)90024-x. [DOI] [PubMed] [Google Scholar]

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