Skip to main content
PMC home page
Journal of Bacteriology logoLink to Journal of Bacteriology
. 1970 Nov;104(2):890–901. doi: 10.1128/jb.104.2.890-901.1970

Protein Composition of the Cell Wall and Cytoplasmic Membrane of Escherichia coli

Carl A Schnaitman 1
PMCID: PMC285073  PMID: 4099097

Abstract

Envelope preparations obtained by passing Escherichia coli cells through a French pressure cell were separated by sucrose density gradient centrifugation into two distinct particulate fractions. The fraction with the higher density was enriched in fragments derived from the cell wall, as indicated by the high content of lipopolysaccharide, the low content of cytochromes, and the similar morphology of the fragments and intact cell walls. The less-dense fraction was enriched in vesicles derived from the cytoplasmic membrane, as indicated by the enrichment of cytochromes, the enzymes lactic and succinic dehydrogenase and nitrate reductase, and the morphological similarity of the vesicles to intact cytoplasmic membrane. Both fractions were rich in phospholipid. The protein composition was compared by mixing the cytoplasmic membrane-enriched fraction from a 3H-labeled culture with the cell wall-enriched fraction from a 14C-labeled culture and examining the resulting mixture by gel electrophoresis. Thirty-four bands of radioactive protein were resolved; of these, 27 were increased two- to fourfold in the cytoplasmic membrane-enriched fraction, whereas 6 were similarly increased in the cell wall-enriched fraction. One of the proteins which is clearly localized in the cell wall is the protein with a molecular weight of 44,000, which is the major component of the envelope. This protein accounted for 70% of the total protein of the cell wall, and its occurrence in the envelope from spheroplasts suggests that it is a structural protein of the outer membranous component of the cell wall.

Full text

PDF
894

Images in this article

893Image
on p.893
896Image
on p.896
897Image
on p.897
899Image
on p.899

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., Rehn K. Chemical characterization, spatial distribution and function of a lipoprotein (murein-lipoprotein) of the E. coli cell wall. The specific effect of trypsin on the membrane structure. Eur J Biochem. 1969 Oct;10(3):426–438. doi: 10.1111/j.1432-1033.1969.tb00707.x. [DOI] [PubMed] [Google Scholar]
  3. Braun V., Sieglin U. The covalent murein-lipoprotein structure of the Escherichia coli cell wall. The attachment site of the lipoprotein on the murein. Eur J Biochem. 1970 Apr;13(2):336–346. doi: 10.1111/j.1432-1033.1970.tb00936.x. [DOI] [PubMed] [Google Scholar]
  4. 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]
  5. COTA-ROBLES E. H. ELECTRON MICROSCOPY OF PLASMOLYSIS IN ESCHERICHIA COLI. J Bacteriol. 1963 Mar;85:499–503. doi: 10.1128/jb.85.3.499-503.1963. [DOI] [PMC free article] [PubMed] [Google Scholar]
  6. 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]
  7. 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]
  8. Kaback H. R., Stadtman E. R. Proline uptake by an isolated cytoplasmic membrane preparation of Escherichia coli. Proc Natl Acad Sci U S A. 1966 Apr;55(4):920–927. doi: 10.1073/pnas.55.4.920. [DOI] [PMC free article] [PubMed] [Google Scholar]
  9. LEUTGEB W., MAASS D., WEIDEL W. HERSTELLUNG UND QUANTITATIVE ZUSAMMENSETZUNG PROTEIN-UND GLUCOSEFREIER STUETZMEMBRANEN (R-LAYERS) AUS COLI-ZELLWAENDEN. Z Naturforsch B. 1963 Dec;18:1062–1064. [PubMed] [Google Scholar]
  10. Levy S. B., Leive L. An equilibrium between two fractions of lipopolysaccharide in Escherichia coli. Proc Natl Acad Sci U S A. 1968 Dec;61(4):1435–1439. doi: 10.1073/pnas.61.4.1435. [DOI] [PMC free article] [PubMed] [Google Scholar]
  11. MURRAY R. G., STEED P., ELSON H. E. THE LOCATION OF THE MUCOPEPTIDE IN SECTIONS OF THE CELL WALL OF ESCHERICHIA COLI AND OTHER GRAM-NEGATIVE BACTERIA. Can J Microbiol. 1965 Jun;11:547–560. doi: 10.1139/m65-072. [DOI] [PubMed] [Google Scholar]
  12. Martin H. H. Bacterial protoplasts--a review. J Theor Biol. 1963 Jul;5(1):1–34. doi: 10.1016/0022-5193(63)90034-1. [DOI] [PubMed] [Google Scholar]
  13. Mergenhagen S. E., Bladen H. A., Hsu K. C. Electron microscopic localization of endotoxic lipopolysaccharide in gram-negative organisms. Ann N Y Acad Sci. 1966 Jun 30;133(2):279–291. doi: 10.1111/j.1749-6632.1966.tb52371.x. [DOI] [PubMed] [Google Scholar]
  14. 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]
  15. Nanninga N. Ultrastructure of the cell envelope of Escherichia coli B after freeze-etching. J Bacteriol. 1970 Jan;101(1):297–303. doi: 10.1128/jb.101.1.297-303.1970. [DOI] [PMC free article] [PubMed] [Google Scholar]
  16. Schnaitman C. A. Examination of the protein composition of the cell envelope of Escherichia coli by polyacrylamide gel electrophoresis. J Bacteriol. 1970 Nov;104(2):882–889. doi: 10.1128/jb.104.2.882-889.1970. [DOI] [PMC free article] [PubMed] [Google Scholar]
  17. Schnaitman C., Greenawalt J. W. Enzymatic properties of the inner and outer membranes of rat liver mitochondria. J Cell Biol. 1968 Jul;38(1):158–175. doi: 10.1083/jcb.38.1.158. [DOI] [PMC free article] [PubMed] [Google Scholar]
  18. Shands J. W. Localization of Somatic Antigen on Gram-Negative Bacteria by Electron Microscopy. J Bacteriol. 1965 Jul;90(1):266–270. doi: 10.1128/jb.90.1.266-270.1965. [DOI] [PMC free article] [PubMed] [Google Scholar]
  19. Strauss J. H., Jr, Burge B. W., Pfefferkorn E. R., Darnell J. E., Jr Identification of the membrane protein and "core" protein of Sindbis virus. Proc Natl Acad Sci U S A. 1968 Feb;59(2):533–537. doi: 10.1073/pnas.59.2.533. [DOI] [PMC free article] [PubMed] [Google Scholar]
  20. TANIGUCHI S., ITAGAKI E. Nitrate reductase of nitrate respiration type from E. coli. I. Solubilization and purification from the particulate system with molecular characterization as a metalloprotein. Biochim Biophys Acta. 1960 Nov 4;44:263–279. doi: 10.1016/0006-3002(60)91562-6. [DOI] [PubMed] [Google Scholar]
  21. WEIDEL W., FRANK H., MARTIN H. H. The rigid layer of the cell wall of Escherichia coli strain B. J Gen Microbiol. 1960 Feb;22:158–166. doi: 10.1099/00221287-22-1-158. [DOI] [PubMed] [Google Scholar]
  22. Weiser M. M., Rothfield L. The reassociation of lipopolysaccharide, phospholipid, and transferase enzymes of the bacterial cell envelope. Isolation of binary and ternary complexes. J Biol Chem. 1968 Mar 25;243(6):1320–1328. [PubMed] [Google Scholar]

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

RESOURCES