Skip to main content
NCBI home page
Journal of Bacteriology logoLink to Journal of Bacteriology
. 1973 Jan;113(1):445–451. doi: 10.1128/jb.113.1.445-451.1973

Freeze-Etching and X-Ray Diffraction of the Isolated Double-Track Layer from the Cell Wall of a Gram-Negative Marine Pseudomonad

A Forge a,1, J W Costerton a, K Ann Kerr a
PMCID: PMC251647  PMID: 4120201

Abstract

The isolated double-track layer of the cell wall of the gram-negative marine pseudomonad studied here contains a cleavage plane. This finding localizes the single cleavage plane of the cell wall and shows that the molecular architecture of this layer provides the lipid-enriched layer which cleaves preferentially in the frozen cell. The observation that the isolated double-track layer of the cell wall is sufficiently ordered at the molecular level to yield a well-defined X-ray diffraction pattern with a d-spacing of 0.44 nm shows that its molecular architecture is very similar to that of true membranes. This specific d-spacing is produced by the highly ordered packing of the hydrophobic portions of phospholipid molecules. Therefore, the double-track layer of the cell wall has been shown, by these two biophysical means, to have a molecular architecture which would allow it to function as the membrane-like “molecular sieve” layer, whose presence has been deduced from physiological data. This layer is important in the retention of cell wall-associated enzymes and in the control of the movement of large molecules through the cell wall.

Full text

PDF
445

Images in this article

447Image
on p.447
447Image
on p.447
448Image
on p.448
449Image
on p.449
450Image
on p.450

Selected References

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

  1. Bayer M. E., Remsen C. C. Structure of Escherichia coli after freeze-etching. J Bacteriol. 1970 Jan;101(1):304–313. doi: 10.1128/jb.101.1.304-313.1970. [DOI] [PMC free article] [PubMed] [Google Scholar]
  2. Braun V., Wolff H. The murein-lipoprotein linkage in the cell wall of Escherichia coli. Eur J Biochem. 1970 Jun;14(2):387–391. doi: 10.1111/j.1432-1033.1970.tb00301.x. [DOI] [PubMed] [Google Scholar]
  3. Burge R. E., Draper J. C. The structure of the cell wall of the Gram-negative bacterium Proteus vulgaris. 3. A lipopolysaccharide "unit membrane". J Mol Biol. 1967 Sep 14;28(2):205–210. doi: 10.1016/s0022-2836(67)80003-2. [DOI] [PubMed] [Google Scholar]
  4. Burge R. E., Draper J. C. The structure of the cell wall of the Gram-negative bacterium Proteus vulgaris. I. An electron microscope and x-ray study. J Mol Biol. 1967 Sep 14;28(2):173–187. doi: 10.1016/s0022-2836(67)80001-9. [DOI] [PubMed] [Google Scholar]
  5. Cerny G., Teuber M. Differential release of periplasmic versus cytoplasmic enzymes from Escherichia coli B by polymixin B. Arch Mikrobiol. 1971;78(2):166–179. doi: 10.1007/BF00424873. [DOI] [PubMed] [Google Scholar]
  6. Cheng K. J., Ingram J. M., Costerton J. W. Interactions of alkaline phosphatase and the cell wall of Pseudomonas aeruginosa. J Bacteriol. 1971 Jul;107(1):325–336. doi: 10.1128/jb.107.1.325-336.1971. [DOI] [PMC free article] [PubMed] [Google Scholar]
  7. Costerton J. W., Forsberg C., Matula T. I., Buckmire F. L., MacLeod R. A. Nutrition and metabolism of marine bacteria. XVI. Formation of protoplasts, spheroplasts, and related forms from a gram-negative marine bacterium. J Bacteriol. 1967 Nov;94(5):1764–1777. doi: 10.1128/jb.94.5.1764-1777.1967. [DOI] [PMC free article] [PubMed] [Google Scholar]
  8. Costerton J. W. The structure and function of the cell envelope of gram-negative bacteria. Rev Can Biol. 1970 Sep;29(3):299–316. [PubMed] [Google Scholar]
  9. Costerton J. W., Thompson J. Induced morphological changes in the stainable layers of the cell envelope of a gram-negative bacterium. Can J Microbiol. 1972 Jun;18(6):937–940. doi: 10.1139/m72-144. [DOI] [PubMed] [Google Scholar]
  10. De Voe I. W., Thompson J., Costerton J. W., MacLeod R. A. Stability and comparative transport capacity of cells, mureinoplasts, and true protoplasts of a gram-negative bacterium. J Bacteriol. 1970 Mar;101(3):1014–1026. doi: 10.1128/jb.101.3.1014-1026.1970. [DOI] [PMC free article] [PubMed] [Google Scholar]
  11. DeVoe I. W., Costerton J. W., MacLeod R. A. Demonstration by freeze-etching of a single cleavage plane in the cell wall of a gram-negative bacterium. J Bacteriol. 1971 May;106(2):659–671. doi: 10.1128/jb.106.2.659-671.1971. [DOI] [PMC free article] [PubMed] [Google Scholar]
  12. Deamer D. W., Branton D. Fracture planes in an ice-bilayer model membrane system. Science. 1967 Nov 3;158(3801):655–657. doi: 10.1126/science.158.3801.655. [DOI] [PubMed] [Google Scholar]
  13. Dreher K. D., Schulman J. H., Anderson O. R., Roels O. A. The stability and structure of mixed lipid monolayers and bilayers. I. Properties of lipid and lipoprotein monolayers on OsO4 solutions and the role of cholesterol, retinol, and tocopherol in stabilizing lecithin monolayers. J Ultrastruct Res. 1967 Aug 30;19(5):586–599. doi: 10.1016/s0022-5320(67)80084-4. [DOI] [PubMed] [Google Scholar]
  14. Engelman D. M. X-ray diffraction studies of phase transitions in the membrane of Mycoplasma laidlawii. J Mol Biol. 1970 Jan 14;47(1):115–117. doi: 10.1016/0022-2836(70)90407-9. [DOI] [PubMed] [Google Scholar]
  15. Fiil A., Branton D. Changes in the plasma membrane of Escherichia coli during magnesium starvation. J Bacteriol. 1969 Jun;98(3):1320–1327. doi: 10.1128/jb.98.3.1320-1327.1969. [DOI] [PMC free article] [PubMed] [Google Scholar]
  16. Finean J. B. Biophysical contributions to membrane structure. Q Rev Biophys. 1969 May;2(1):1–23. doi: 10.1017/s0033583500000779. [DOI] [PubMed] [Google Scholar]
  17. Fleischer S., Fleischer B., Stoeckenius W. Fine structure of lipid-depleted mitochondria. J Cell Biol. 1967 Jan;32(1):193–208. doi: 10.1083/jcb.32.1.193. [DOI] [PMC free article] [PubMed] [Google Scholar]
  18. Forsberg C. W., Costerton J. W., Macleod R. A. Quantitation, chemical characteristics, and ultrastructure of the three outer cell wall layers of a gram-negative bacterium. J Bacteriol. 1970 Dec;104(3):1354–1368. doi: 10.1128/jb.104.3.1354-1368.1970. [DOI] [PMC free article] [PubMed] [Google Scholar]
  19. Forsberg C. W., Costerton J. W., Macleod R. A. Separation and localization of cell wall layers of a gram-negative bacterium. J Bacteriol. 1970 Dec;104(3):1338–1353. doi: 10.1128/jb.104.3.1338-1353.1970. [DOI] [PMC free article] [PubMed] [Google Scholar]
  20. Johnson S. M., Bangham A. D., Hill M. W., Korn E. D. Single bilayer liposomes. Biochim Biophys Acta. 1971 Jun 1;233(3):820–826. doi: 10.1016/0005-2736(71)90184-2. [DOI] [PubMed] [Google Scholar]
  21. Margulis L. Symbiosis and evolution. Sci Am. 1971 Aug;225(2):48–57. doi: 10.1038/scientificamerican0871-48. [DOI] [PubMed] [Google Scholar]
  22. Martin E. L., MacLeod R. A. Isolation and chemical composition of the cytoplasmic membrane of a gram-negative bacterium. J Bacteriol. 1971 Mar;105(3):1160–1167. doi: 10.1128/jb.105.3.1160-1167.1971. [DOI] [PMC free article] [PubMed] [Google Scholar]
  23. Nisonson I., Tannenbaum M., Neu H. C. Surface localization of Escherichia coli 5'-nucleotidase by electron microscopy. J Bacteriol. 1969 Nov;100(2):1083–1090. doi: 10.1128/jb.100.2.1083-1090.1969. [DOI] [PMC free article] [PubMed] [Google Scholar]
  24. Payne J. W., Gilvarg C. Size restriction on peptide utilization in Escherichia coli. J Biol Chem. 1968 Dec 10;243(23):6291–6299. [PubMed] [Google Scholar]
  25. Pinto da Silva P., Branton D. Membrane splitting in freeze-ethching. Covalently bound ferritin as a membrane marker. J Cell Biol. 1970 Jun;45(3):598–605. doi: 10.1083/jcb.45.3.598. [DOI] [PMC free article] [PubMed] [Google Scholar]
  26. REYNOLDS E. S. The use of lead citrate at high pH as an electron-opaque stain in electron microscopy. J Cell Biol. 1963 Apr;17:208–212. doi: 10.1083/jcb.17.1.208. [DOI] [PMC free article] [PubMed] [Google Scholar]
  27. Reiss-Husson F., Luzzati V. Phase transitions in lipids in relation to the structure of membranes. Adv Biol Med Phys. 1967;11:87–107. doi: 10.1016/b978-1-4832-3107-5.50007-8. [DOI] [PubMed] [Google Scholar]
  28. Schnaitman C. A. Effect of ethylenediaminetetraacetic acid, Triton X-100, and lysozyme on the morphology and chemical composition of isolate cell walls of Escherichia coli. J Bacteriol. 1971 Oct;108(1):553–563. doi: 10.1128/jb.108.1.553-563.1971. [DOI] [PMC free article] [PubMed] [Google Scholar]
  29. Singh A. P., Cheng K. J., Costerton J. W., Idziak E. S., Ingram J. M. Sensitivity of normal and mutant strains of Escherichia coli to actinomycin-D. Can J Microbiol. 1972 Jun;18(6):909–915. doi: 10.1139/m72-139. [DOI] [PubMed] [Google Scholar]
  30. van Gool A. P., Nanninga N. Fracture faces in the cell envelope of Escherichia coli. J Bacteriol. 1971 Oct;108(1):474–481. doi: 10.1128/jb.108.1.474-481.1971. [DOI] [PMC free article] [PubMed] [Google Scholar]

Articles from Journal of Bacteriology are provided here courtesy of American Society for Microbiology (ASM)

RESOURCES