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. 1971 Mar;105(3):1184–1199. doi: 10.1128/jb.105.3.1184-1199.1971

Dissociation and Reassembly of Escherichia coli Outer Membrane and of Lipopolysaccharide, and Their Reassembly onto Flagellar Basal Bodies

M L DePamphilis 1
PMCID: PMC248550  PMID: 4100836

Abstract

Purified lipopolysaccharide vesicles dissociate when treated with ethylenediaminetetraacetic acid (EDTA) and then reassemble when dialyzed against Mg2+. Purified outer, lipopolysaccharide membrane (L membrane) is partially dissociated by treatment with EDTA and fully dissociated upon further treatment with Triton X-100. Both the partially and fully dissociated L membrane can be reassembled by dialysis against Mg2+. Reassembly of lipopolysaccharide or L membrane in the presence of intact flagella results in specific attachment of flagellar basal bodies to vesicles via the L and sometimes the M ring. Lipopolysaccharide and L membrane appear to be composed of substructures bound together by both Mg2+ (divalent cation)-mediated and hydrophobic bonds.

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Selected References

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  1. De Petris S. Ultrastructure of the cell wall of Escherichia coli and chemical nature of its constituent layers. J Ultrastruct Res. 1967 Jul;19(1):45–83. doi: 10.1016/s0022-5320(67)80059-5. [DOI] [PubMed] [Google Scholar]
  2. DePamphilis M. L., Adler J. Attachment of flagellar basal bodies to the cell envelope: specific attachment to the outer, lipopolysaccharide membrane and the cyoplasmic membrane. J Bacteriol. 1971 Jan;105(1):396–407. doi: 10.1128/jb.105.1.396-407.1971. [DOI] [PMC free article] [PubMed] [Google Scholar]
  3. DePamphilis M. L., Adler J. Fine structure and isolation of the hook-basal body complex of flagella from Escherichia coli and Bacillus subtilis. J Bacteriol. 1971 Jan;105(1):384–395. doi: 10.1128/jb.105.1.384-395.1971. [DOI] [PMC free article] [PubMed] [Google Scholar]
  4. DePamphilis M. L., Adler J. Purification of intact flagella from Escherichia coli and Bacillus subtilis. J Bacteriol. 1971 Jan;105(1):376–383. doi: 10.1128/jb.105.1.376-383.1971. [DOI] [PMC free article] [PubMed] [Google Scholar]
  5. ELBEIN A. D., HEATH E. C. THE BIOSYNTHESIS OF CELL WALL LIPOPOLYSACCHARIDE IN ESCHERICHIA COLI. I. THE BIOCHEMICAL PROPERTIES OF A URIDINE DIPHOSPHATE GALACTOSE 4-EPIMERASELESS MUTANT. J Biol Chem. 1965 May;240:1919–1925. [PubMed] [Google Scholar]
  6. Engelman D. M., Morowitz H. J. Characterization of the plasma membrane of Mycoplasma laidlawii. IV. Structure and composition of membrane and aggregated components. Biochim Biophys Acta. 1968 Apr 29;150(3):385–396. doi: 10.1016/0005-2736(68)90137-5. [DOI] [PubMed] [Google Scholar]
  7. Frank H., Dekegel D. Electron microscopical studies on the localisation of the different components of cell walls of gram-negative bacteria. Folia Microbiol (Praha) 1967;12(3):227–233. doi: 10.1007/BF02868736. [DOI] [PubMed] [Google Scholar]
  8. Knox K. W., Vesk M., Work E. Relation between excreted lipopolysaccharide complexes and surface structures of a lysine-limited culture of Escherichia coli. J Bacteriol. 1966 Oct;92(4):1206–1217. doi: 10.1128/jb.92.4.1206-1217.1966. [DOI] [PMC free article] [PubMed] [Google Scholar]
  9. 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]
  10. Leive L., Shovlin V. K., Mergenhagen S. E. Physical, chemical, and immunological properties of lipopolysaccharide released from Escherichia coli by ethylenediaminetetraacetate. J Biol Chem. 1968 Dec 25;243(24):6384–6391. [PubMed] [Google Scholar]
  11. McIntire F. C., Sievert H. W., Barlow G. H., Finley R. A., Lee A. Y. Chemical, physical, biological properties of a lipopolysaccharide from Escherichia coli K-235. Biochemistry. 1967 Aug;6(8):2363–2372. doi: 10.1021/bi00860a011. [DOI] [PubMed] [Google Scholar]
  12. Miura T., Mizushima S. Separation and properties of outer and cytoplasmic membranes in Escherichia coli. Biochim Biophys Acta. 1969;193(2):268–276. doi: 10.1016/0005-2736(69)90188-6. [DOI] [PubMed] [Google Scholar]
  13. OSBORN M. J., ROSEN S. M., ROTHFIELD L., HORECKER B. L. Biosynthesis of bacterial lipopolysaccharide. I. Enzymatic incorporation of galactose in a mutant strain of Salmonella. Proc Natl Acad Sci U S A. 1962 Oct 15;48:1831–1838. doi: 10.1073/pnas.48.10.1831. [DOI] [PMC free article] [PubMed] [Google Scholar]
  14. OSBORN M. J. STUDIES ON THE GRAM-NEGATIVE CELL WALL. I. EVIDENCE FOR THE ROLE OF 2-KETO- 3-DEOXYOCTONATE IN THE LIPOPOLYSACCHARIDE OF SALMONELLA TYPHIMURIUM. Proc Natl Acad Sci U S A. 1963 Sep;50:499–506. doi: 10.1073/pnas.50.3.499. [DOI] [PMC free article] [PubMed] [Google Scholar]
  15. Osborn M. J. Structure and biosynthesis of the bacterial cell wall. Annu Rev Biochem. 1969;38:501–538. doi: 10.1146/annurev.bi.38.070169.002441. [DOI] [PubMed] [Google Scholar]
  16. Rapin A. M., Mayer H. Complex polysaccharides in different strains of Escherichia coli K-12. Ann N Y Acad Sci. 1966 Jun 30;133(2):425–437. doi: 10.1111/j.1749-6632.1966.tb52381.x. [DOI] [PubMed] [Google Scholar]
  17. Razin S., Ne'eman Z., Ohad I. Selective reaggregation of solubilized Mycoplasma-membrane proteins and the kinetics of membrane reformation. Biochim Biophys Acta. 1969;193(2):277–293. doi: 10.1016/0005-2736(69)90189-8. [DOI] [PubMed] [Google Scholar]
  18. Ribi E., Anacker R. L., Brown R., Haskins W. T., Malmgren B., Milner K. C., Rudbach J. A. Reaction of endotoxin and surfactants. I. Physical and biological properties of endotoxin treated with sodium deoxycholate. J Bacteriol. 1966 Nov;92(5):1493–1509. doi: 10.1128/jb.92.5.1493-1509.1966. [DOI] [PMC free article] [PubMed] [Google Scholar]
  19. Rothfield L., Horne R. W. Reassociation of purified lipopolysaccharide and phospholipid of the bacterial cell envelope: electron microscopic and monolayer studies. J Bacteriol. 1967 May;93(5):1705–1721. doi: 10.1128/jb.93.5.1705-1721.1967. [DOI] [PMC free article] [PubMed] [Google Scholar]
  20. Shands J. W., Jr, Graham J. A., Nath K. The morphologic structure of isolated bacterial lipopolysaccharide. J Mol Biol. 1967 Apr 14;25(1):15–21. doi: 10.1016/0022-2836(67)90275-6. [DOI] [PubMed] [Google Scholar]
  21. Taylor A., Knox K. W., Work E. Chemical and biological properties of an extracellular lipopolysaccharide from Escherichia coli grown under lysine-limiting conditions. Biochem J. 1966 Apr;99(1):53–61. doi: 10.1042/bj0990053. [DOI] [PMC free article] [PubMed] [Google Scholar]
  22. WEISSBACH A., HURWITZ J. The formation of 2-keto-3-deoxyheptonic acid in extracts of Escherichia coli B. I. Identification. J Biol Chem. 1959 Apr;234(4):705–709. [PubMed] [Google Scholar]
  23. Work E. The chemistry and morphology of surface structures of lysine-limited Escherichia coli. Folia Microbiol (Praha) 1967;12(3):220–226. doi: 10.1007/BF02868735. [DOI] [PubMed] [Google Scholar]

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