Skip to main content
NCBI home page
Journal of Bacteriology logoLink to Journal of Bacteriology
. 1974 Mar;117(3):1335–1340. doi: 10.1128/jb.117.3.1335-1340.1974

Chemical Composition and Ultrastructure of Native and Reaggregated Membranes from Protoplasts of Bacillus cereus

Teofila Cabrera Beaman 1, H Stuart Pankratz 1, Philipp Gerhardt 1
PMCID: PMC246618  PMID: 4205199

Abstract

Conditions were defined for producing protoplasts with lysozyme and isolating the protoplast membranes from cells of Bacillus cereus T harvested late in the exponential growth phase just before sporogenesis. The membranes contained approximately 60% protein, 30% lipid, 6% carbohydrate, and 1% ribonucleic acid. Seventeen proteins were distinguished by molecular size in the membrane solubilized with sodium dodecyl sulfate, and 12 in that with phenol and acetic acid. The lipid fraction consisted of neutral lipids (28%) and phospholipids (72%). Four phospholipids were identified: diphosphatidyl glycerol, phosphatidyl ethanolamine, phosphatidyl glycerol, and lysophosphatidyl ethanolamine. Eighteen fatty acids were identified, with a predominance of branched C15 and C17 and of normal C16 acids. The carbohydrate fraction consisted of neutral hexoses. A clear supernatant solution from the solubilized preparation became reaggregated into membrane by dialysis in the presence of MgCl2. The reaggregated membrane had the same main components as the native membrane, but the amount and ratio of protein and lipid depended on the buffer and the MgCl2 concentration. By electron microscopy, the reaggregated membranes appeared as vesicles or sheets, depending on the MgCl2 concentration. Hexagonal lattices were occasionally detected in the negatively stained ultrastructure of both native and reaggregated membrane fragments.

Full text

PDF
1335

Images in this article

1338Image
on p.1338

Selected References

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

  1. BARTLETT G. R. Phosphorus assay in column chromatography. J Biol Chem. 1959 Mar;234(3):466–468. [PubMed] [Google Scholar]
  2. Beaman T. C., Pankratz H. S., Gerhardt P. Paracrystalline sheets reaggregated from solubilized exosporium of Bacillus cereus. J Bacteriol. 1971 Jul;107(1):320–324. doi: 10.1128/jb.107.1.320-324.1971. [DOI] [PMC free article] [PubMed] [Google Scholar]
  3. Benedetti E. L., Emmelot P. Hexagonal array of subunits in tight junctions separated from isolated rat liver plasma membranes. J Cell Biol. 1968 Jul;38(1):15–24. doi: 10.1083/jcb.38.1.15. [DOI] [PMC free article] [PubMed] [Google Scholar]
  4. Bishop D. G., Rutberg L., Samuelsson B. The chemical composition of the cytoplasmic membrane of Bacillus subtilis. Eur J Biochem. 1967 Nov;2(4):448–453. doi: 10.1111/j.1432-1033.1967.tb00158.x. [DOI] [PubMed] [Google Scholar]
  5. Butler T. F., Smith G. L., Grula E. A. Bacterial cell membranes. I. Reaggregation of membrane subunits from Micrococcus lysodeikticus. Can J Microbiol. 1967 Nov;13(11):1471–1479. doi: 10.1139/m67-195. [DOI] [PubMed] [Google Scholar]
  6. Dunker A. K., Rueckert R. R. Observations on molecular weight determinations on polyacrylamide gel. J Biol Chem. 1969 Sep 25;244(18):5074–5080. [PubMed] [Google Scholar]
  7. FOLCH J., LEES M., SLOANE STANLEY G. H. A simple method for the isolation and purification of total lipides from animal tissues. J Biol Chem. 1957 May;226(1):497–509. [PubMed] [Google Scholar]
  8. Felix J. A., Lundgren D. G. Electron transport system associated with membranes of Bacillus cereus during vegetative growth and sporulation. J Bacteriol. 1973 Aug;115(2):552–559. doi: 10.1128/jb.115.2.552-559.1973. [DOI] [PMC free article] [PubMed] [Google Scholar]
  9. GERHARDT P., RIBI E. ULTRASTRUCTURE OF THE EXOSPORIUM ENVELOPING SPORES OF BACILLUS CEREUS. J Bacteriol. 1964 Dec;88:1774–1789. doi: 10.1128/jb.88.6.1774-1789.1964. [DOI] [PMC free article] [PubMed] [Google Scholar]
  10. HASHIMOTO T., BLACK S. H., GERHARDT P. Development of fine structure, thermostability, and dipicolinate during sporogenesis in a bacillus. Can J Microbiol. 1960 Apr;6:203–212. doi: 10.1139/m60-022. [DOI] [PubMed] [Google Scholar]
  11. Hicks R. M., Ketterer B. Isolation of the plasma membrane of the luminal surface of rat bladder epithelium, and the occurrence of a hexagonal lattice of subunits both in negatively stained whole mounts and in sectioned membranes. J Cell Biol. 1970 Jun;45(3):542–553. doi: 10.1083/jcb.45.3.542. [DOI] [PMC free article] [PubMed] [Google Scholar]
  12. LAW J. H., SLEPECKY R. A. Assay of poly-beta-hydroxybutyric acid. J Bacteriol. 1961 Jul;82:33–36. doi: 10.1128/jb.82.1.33-36.1961. [DOI] [PMC free article] [PubMed] [Google Scholar]
  13. 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]
  14. Matz L. L., Beaman T. C., Gerhardt P. Chemical composition of exosporium from spores of Bacillus cereus. J Bacteriol. 1970 Jan;101(1):196–201. doi: 10.1128/jb.101.1.196-201.1970. [DOI] [PMC free article] [PubMed] [Google Scholar]
  15. Patch C. T., Landman O. E. Comparison of the biochemistry and rates of synthesis of mesosomal and peripheral membranes in Bacillus subtilis. J Bacteriol. 1971 Jul;107(1):345–357. doi: 10.1128/jb.107.1.345-357.1971. [DOI] [PMC free article] [PubMed] [Google Scholar]
  16. RONDLE C. J., MORGAN W. T. The determination of glucosamine and galactosamine. Biochem J. 1955 Dec;61(4):586–589. doi: 10.1042/bj0610586. [DOI] [PMC free article] [PubMed] [Google Scholar]
  17. Razin S., Boschwitz C. The membrane of the Streptobacillus moniliformis L-phase. J Gen Microbiol. 1968 Nov;54(1):21–32. doi: 10.1099/00221287-54-1-21. [DOI] [PubMed] [Google Scholar]
  18. Razin S., Morowitz H. J., Terry T. M. Membrane subunits of Mycoplasma laidlawii and their assembly to membranelike structures. Proc Natl Acad Sci U S A. 1965 Jul;54(1):219–225. doi: 10.1073/pnas.54.1.219. [DOI] [PMC free article] [PubMed] [Google Scholar]
  19. Rottem S., Stein O., Razin S. Reassembly of Mycoplasma membranes disaggregated by detergents. Arch Biochem Biophys. 1968 Apr;125(1):46–56. doi: 10.1016/0003-9861(68)90637-1. [DOI] [PubMed] [Google Scholar]
  20. Salton M. R., Freer J. H. Composition of the membranes isolated from several Gram-positive bacteria. Biochim Biophys Acta. 1965 Oct 18;107(3):531–538. doi: 10.1016/0304-4165(65)90197-2. [DOI] [PubMed] [Google Scholar]
  21. TOENNIES G., KOLB J. J. CARBOHYDRATE ANALYSIS OF BACTERIAL SUBSTANCES BY A NEW ANTHRONE PROCEDURE. Anal Biochem. 1964 May;8:54–69. doi: 10.1016/0003-2697(64)90168-x. [DOI] [PubMed] [Google Scholar]
  22. WEIBULL C., BERGSTROM L. The chemical nature of the cytoplasmic membrane and cell wall of Bacillus megaterium, strain M. Biochim Biophys Acta. 1958 Nov;30(2):340–351. doi: 10.1016/0006-3002(58)90059-3. [DOI] [PubMed] [Google Scholar]
  23. Weber K., Osborn M. The reliability of molecular weight determinations by dodecyl sulfate-polyacrylamide gel electrophoresis. J Biol Chem. 1969 Aug 25;244(16):4406–4412. [PubMed] [Google Scholar]
  24. Yamaguchi T., Tamura G., Arima K. Substructure of the cytoplasmic membrane of Bacillus megaterium. I. Method for the fractionation of "Ghosts". J Bacteriol. 1967 Jan;93(1):483–489. doi: 10.1128/jb.93.1.483-489.1967. [DOI] [PMC free article] [PubMed] [Google Scholar]
  25. Yudkin M. D. Isolation and analysis of the protoplast membrane of Bacillus megaterium. Biochem J. 1966 Mar;98(3):923–928. doi: 10.1042/bj0980923. [DOI] [PMC free article] [PubMed] [Google Scholar]

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

RESOURCES