1887

Abstract

SUMMARY: Electron microscopy on thin sections of three different extremely halophilic Halobacterium species showed that their cell envelopes were of similar general construction: an inner membrane and an outer layer. The outer layer stains most strongly in the outermost part. When the NaCl concentration of the environment was lowered from the optimal of 4·3 to 2·2 the outer layer of the cell envelope of strain 1 became frayed; in many cells a release of material from the outer layer appeared to take place. When the cells were exposed to distilled water the outer layer of the envelope appeared to dissolve completely and the cell membrane disintegrated into tiny flakes. Fragments of the cell envelope produced by mechanical disintegration of the cells in 4·3 -NaCl formed closed vesicles very rapidly; some of the cytoplasmic material became trapped inside the vesicles. Detergents appeared to slow down the closing of the vesicles and also to cause a release of material from the outer layer of the cell envelope. The cell envelope vesicles were mainly composed of protein and lipid; their content of amino sugar was low compared with the cell envelope of other Gram-negative bacteria. The cell envelope vesicle also contained nucleic acids; most of these were probably parts of the cytoplasmic material trapped inside the vesicles. The amino acid composition showed that the protein of the cell envelope vesicles was quite acidic, consistent with the contention that high concentrations of sodium ions stabilize the cell envelopes of these organisms by neutralizing the negative charges of the protein. Upon centrifugation at high speed of the lysate obtained by dialysis of the cell envelope vesicles against distilled water, the fragments of the cell membrane sedimented whereas most of the protein, presumably from the outer layer of the cell envelope, stayed in the supernatant fraction. Carotenoids and cytochromes were contained in the sediment with the membrane fragments. Most of the amino sugar-containing components stayed in the supernatant fraction; in the presence of 10–25 salt most of the amino sugar-containing components sediment with the membrane fragments.

Loading

Article metrics loading...

/content/journal/micro/10.1099/00221287-55-3-325
1969-03-01
2024-12-13
Loading full text...

Full text loading...

/deliver/fulltext/micro/55/3/mic-55-3-325.html?itemId=/content/journal/micro/10.1099/00221287-55-3-325&mimeType=html&fmt=ahah

References

  1. Albertson P.-Å. 1960 Partition of Cell Particles and Macromolecules Stockholm: Almqvist and Wiksell;
    [Google Scholar]
  2. Brown A. D. 1963; The peripheral structure of Gram-negative bacteria IV. The cation-sensitive dissolution of the cell membrane of the halophilic bacterium, Halobacterium halobium . Biochim. biophys. Acta 75:425
    [Google Scholar]
  3. Brown A. D., Shorey C. D. 1963; The cell envelopes of two extremely halophilic bacteria. J. cell Biol 18:681
    [Google Scholar]
  4. Brown A. D., Shorey C. D., Turner H. P. 1965; An alternative method of isolating the membrane of a halophilic bacterium. J. gen. Microbiol 41:225
    [Google Scholar]
  5. Cho K. Y., Doy C. H., Mercer E. H. 1967; Ultrastructure of the obligate halophilic bacterium Halobacterium halobium . J. Bact 94:196
    [Google Scholar]
  6. Folch J., Lees M., Stanley G. H. S. 1957; A simple method for the isolation and purification of total lipids from animal tissues. J. biol. Chem 226:497
    [Google Scholar]
  7. Glauert A. M., Thornley M. J. 1966; Glutaraldehyde fixation of Gram-negative bacteria. Jl R. microsc. Soc 85:449
    [Google Scholar]
  8. Holmes P. K., Dundas I. E. D., Halvorson H. O. 1965; Halophilic enzymes in cell-free extracts of Halobacterium salinarium . J. Bact 90:1159
    [Google Scholar]
  9. Howe J. M., Featherston W. R., Stadelman W. J., Banwart G. J. 1965; Amino acid composition of certain bacterial cell-wall proteins. Appl. Microbiol 13:650
    [Google Scholar]
  10. Kellenberger E., Ryter A., Séchaud J. 1958; Electron microscope study of DNA-containing plasma. J. biophys. biochem. Cytol 4:671
    [Google Scholar]
  11. Kushner D. J., Onishi H. 1966; Contribution of protein and lipid components to the salt response of envelopes of an extremely halophilic bacterium. J. Bact 91:653
    [Google Scholar]
  12. Kushner D. J., Bayley S. T., Boring J., Kates M., Gibbons N. E. 1964; Morphological and chemical properties of cell envelopes of the extreme halophile, Halobacterium cutirubrum . Can. J. Microbiol 10:483
    [Google Scholar]
  13. Larsen H. 1962; Halophilism. In The Bacteria Gunsalus I. C., Stanier R. Y. Ed. by 4 p. 297 New York: Academic Press;
    [Google Scholar]
  14. Larsen H. 1967; Biochemical aspects of extreme halophilism. In Advances in Microbial Physiology Rose A. H., Wilkinson J. F. Ed. by 1 p. 97 London: Academic Press;
    [Google Scholar]
  15. Larsen H., Omang S., Steensland H. 1967; On the gas vacuoles of the halobacteria. Arch. Mikrobiol 59:197
    [Google Scholar]
  16. Layne E. 1957; Spectrophotometric and turbidimetric methods for measuring proteins. In Meth. Enzym 3:447
    [Google Scholar]
  17. Liaaen Jensen S. 1960; Bacterial carotenoids. VI. A note on the constitution of bacterioruberine a. Acta chem. scand 14:950
    [Google Scholar]
  18. McClare C. W. F. 1967; Bonding between proteins and lipids in the envelopes of Halobacterium halobium . Nature; Lond: 216766
    [Google Scholar]
  19. Mohr V., Larsen H. 1963; On the structural transformation and lysis of Halobacterium salinarium in hypotonic and isotonic solutions. J. gen. Microbiol 31:267
    [Google Scholar]
  20. Moore S. 1963; On the determination of cystine as cysteic acid. J. biol. Chem 238:235
    [Google Scholar]
  21. Moore S., Spackman D. H., Stein W. H. 1958; Chromatography of amino acids on sulphonated polystyrene resins. Analyt. Chem 30:1185
    [Google Scholar]
  22. Noltmann E. A., Mahowald T. A., Kuby S. A. 1962; Studies on adenosine triphosphate transphosphorylases. J. biol. Chem 237:1146
    [Google Scholar]
  23. Onishi H., Kushner D. J. 1966; Mechanism of dissolution of envelopes of the extreme halophile Halobacterium cutirubrum . J. Bact 91:646
    [Google Scholar]
  24. Penso G. 1947; II rosso dei bacallari-Etiologia, commestibilità, bonifica e prevenzione. Rc. 1st. sup. Sanità 10:563
    [Google Scholar]
  25. Reiner M. 1941 Manual of Clinical Chemistry New York: Interscience;
    [Google Scholar]
  26. Rhuland L. E., Work E., Denman R. F., Hoare D. S. 1955; The behaviour of the isomers of α,ɛ-diaminopimelic acid on paper chromatograms. J. Am. chem. Soc 77:4844
    [Google Scholar]
  27. Rondle C. J. M., Morgan W. T. J. 1955; The determination of glucosamine and galactosamine. Biochem. J 61:586
    [Google Scholar]
  28. Salton M. R. J. 1967; Structure and function of bacterial cell membranes. A. Rev. Microbiol 21:417
    [Google Scholar]
  29. Steensland H. 1967; Electron microscope studies of the surface layers of halobacteria. Abstr. 4th Meeting Federation Europ. Biochem. Soc p. 49 Oslo: Universitetsforlaget;
    [Google Scholar]
  30. Stoeckenius W., Rowen R. 1966; Fine structure of the cell envelope and internal membranes of Halobacterium halobium . Abstr. 6th int. Congr. Electron Microscopy, Kyoto 2 p. 273 Tokyo: Maruzen Co;
    [Google Scholar]
  31. Stoeckenius W., Rowen R. 1967; A morphological study of Halobacterium halobium and its lysis in media of low salt concentration. J. cell Biol 34:365
    [Google Scholar]
  32. Tristram G. R., Smith R. H. 1962; The amino acid composition of some purified proteins. In Advances in Protein Chemistry Anfinsen C. B., Anson M. L., Edsall J. T. Ed. by 18 p. 227 New York and London: Academic Press;
    [Google Scholar]
  33. Yemm E. W., Cocking E. C. 1955; The determination of amino-acids with ninhydrin. Analyst, Lond 80:209
    [Google Scholar]
/content/journal/micro/10.1099/00221287-55-3-325
Loading
/content/journal/micro/10.1099/00221287-55-3-325
Loading

Data & Media loading...

This is a required field
Please enter a valid email address
Approval was a Success
Invalid data
An Error Occurred
Approval was partially successful, following selected items could not be processed due to error