Abstract
Chemical analysis of fractions of the cell envelope of Acinetobacter sp. strain MJT/F5/199A, prepared by breakage in the French press and removal of plasma membranes, followed by sequential treatment with lysozyme and with papain, confirmed the existence of layers previously identified by electron microscopy. Outside the plasma membrane and periplasmic space, the envelope is composed of (i) a peptidoglycan-containing dense layer, (ii) an intermediate layer, (iii) a lipopolysaccharide-containing outer membrane, and (iv) an ordered array of protein subunits. A small amount of carbohydrate (3%) is found associated with protein in the fraction containing both the surface subunits and the intermediate layer. The papain-treated outer membranes contain 67% protein, 24% lipid, together with 11% lipopolysaccharide, and about 6% of non-lipopolysaccharide hexosamine. Lipid is located only in the papain-treated outer-membrane and is mainly phospholipid: 29% phosphatidyl glycerol, 30% phosphatidyl ethanolamine, and 40% cardiolipin. The principal fatty acid is C18:1. Significant amounts of alcohols16:1 and alcohols18:1, which are found in Acinetobacter waxes, were recovered from the outer membrane.
Full text
PDFImages in this article
Selected References
These references are in PubMed. This may not be the complete list of references from this article.
- AMENTA J. S. A RAPID CHEMICAL METHOD FOR QUANTIFICATION OF LIPIDS SEPARATED BY THIN-LAYER CHROMATOGRAPHY. J Lipid Res. 1964 Apr;5:270–272. [PubMed] [Google Scholar]
- AMINOFF D. Methods for the quantitative estimation of N-acetylneuraminic acid and their application to hydrolysates of sialomucoids. Biochem J. 1961 Nov;81:384–392. doi: 10.1042/bj0810384. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Adams G. A., Kates M., Shaw D. H., Yaguchi M. Studies on the chemical constitution of cell-wall lipopolysaccharides from Neisseria perflava. Can J Biochem. 1968 Oct;46(10):1175–1184. doi: 10.1139/o68-176. [DOI] [PubMed] [Google Scholar]
- Adams G. A., Quadling C., Yaguchi M., Tornabene T. G. The chemical composition of cell-wall lipopolysaccharides from Moraxella duplex and Micrococcus calco-aceticus. Can J Microbiol. 1970 Jan;16(1):1–8. doi: 10.1139/m70-001. [DOI] [PubMed] [Google Scholar]
- Adams G. A. Structural investigations on a cell-wall lipopolysaccharide from Neisseria sicca. Can J Biochem. 1971 Feb;49(2):243–250. doi: 10.1139/o71-035. [DOI] [PubMed] [Google Scholar]
- Bayer M. E., Anderson T. F. The surface structure of Escherichia coli. Proc Natl Acad Sci U S A. 1965 Dec;54(6):1592–1599. doi: 10.1073/pnas.54.6.1592. [DOI] [PMC free article] [PubMed] [Google Scholar]
- 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]
- 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]
- Buckmire F. L., Murray R. G. Studies on the cell wall of Spirillum serpens. 1. Isolation and partial purification of the outermost cell wall layer. Can J Microbiol. 1970 Oct;16(10):1011–1022. doi: 10.1139/m70-171. [DOI] [PubMed] [Google Scholar]
- DISCHE Z. Qualitative and quantitative colorimetric determination of heptoses. J Biol Chem. 1953 Oct;204(2):983–997. [PubMed] [Google Scholar]
- Davison P. K., Young R. Gas chromatography of carbohydrates. The quantitative determination of the free sugars of plants as their trimethylsilyl ethers. J Chromatogr. 1969 Apr 22;41(1):12–21. doi: 10.1016/0021-9673(64)80093-5. [DOI] [PubMed] [Google Scholar]
- 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]
- 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]
- Gallagher I. H. Occurrence of waxes in Acinetobacter. J Gen Microbiol. 1971 Oct;68(2):245–247. doi: 10.1099/00221287-68-2-245. [DOI] [PubMed] [Google Scholar]
- Glauert A. M., Thornley M. J. The topography of the bacterial cell wall. Annu Rev Microbiol. 1969;23:159–198. doi: 10.1146/annurev.mi.23.100169.001111. [DOI] [PubMed] [Google Scholar]
- HANES C. S., ISHERWOOD F. A. Separation of the phosphoric esters on the filter paper chromatogram. Nature. 1949 Dec 31;164(4183):1107-12, illust. doi: 10.1038/1641107a0. [DOI] [PubMed] [Google Scholar]
- IBBOTT F. A., ABRAMS A. THE PHOSPHOLIPIDS IN MEMBRANE GHOSTS FROM STREPTOCOCCUS FAECALIS PROTOPLASTS. Biochemistry. 1964 Dec;3:2008–2012. doi: 10.1021/bi00900a039. [DOI] [PubMed] [Google Scholar]
- Kennedy J. F. Liquid scintillation counting medium for aqueous samples. Experientia. 1969 Oct 15;25(10):1120–1120. doi: 10.1007/BF01901473. [DOI] [PubMed] [Google Scholar]
- Knox K. W., Cullen J., Work E. An extracellular lipopolysaccharide-phospholipid-protein complex produced by Escherichia coli grown under lysine-limiting conditions. Biochem J. 1967 Apr;103(1):192–201. doi: 10.1042/bj1030192. [DOI] [PMC free article] [PubMed] [Google Scholar]
- 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]
- MORRISON W. R., SMITH L. M. PREPARATION OF FATTY ACID METHYL ESTERS AND DIMETHYLACETALS FROM LIPIDS WITH BORON FLUORIDE--METHANOL. J Lipid Res. 1964 Oct;5:600–608. [PubMed] [Google Scholar]
- 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]
- Miura T., Mizushima S. Separation by density gradient centrifugation of two types of membranes from spheroplast membrane of Escherichia coli K12. Biochim Biophys Acta. 1968 Jan 3;150(1):159–161. doi: 10.1016/0005-2736(68)90020-5. [DOI] [PubMed] [Google Scholar]
- Osborn M. J., Gander J. E., Parisi E., Carson J. Mechanism of assembly of the outer membrane of Salmonella typhimurium. Isolation and characterization of cytoplasmic and outer membrane. J Biol Chem. 1972 Jun 25;247(12):3962–3972. [PubMed] [Google Scholar]
- Pate J. L., Ordal E. J. The fine structure of Chondrococcus columnaris. 3. The surface layers of Chondrococcus columnaris. J Cell Biol. 1967 Oct;35(1):37–51. doi: 10.1083/jcb.35.1.37. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Randle C. L., Albro P. W., Dittmer J. C. The phosphoglyceride composition of Gram-negative bacteria and the changes in composition during growth. Biochim Biophys Acta. 1969;187(2):214–220. doi: 10.1016/0005-2760(69)90030-7. [DOI] [PubMed] [Google Scholar]
- Shaw N. Bacterial glycolipids. Bacteriol Rev. 1970 Dec;34(4):365–377. doi: 10.1128/br.34.4.365-377.1970. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Shaw N. The detection of lipids on thin-layer chromatograms with the periodate-Schiff reagents. Biochim Biophys Acta. 1968 Oct 22;164(2):435–436. doi: 10.1016/0005-2760(68)90171-9. [DOI] [PubMed] [Google Scholar]
- Thornley M. J., Glauert A. M. Fine structure and radiation resistance in Acinetobacter: studies on a resistant strain. J Cell Sci. 1968 Jun;3(2):273–294. doi: 10.1242/jcs.3.2.273. [DOI] [PubMed] [Google Scholar]
- Thornley M. J., Glauert A. M., Sleytr U. B. Isolation of outer membranes with an ordered array of surface subunits from Acinetobacter. J Bacteriol. 1973 Jun;114(3):1294–1308. doi: 10.1128/jb.114.3.1294-1308.1973. [DOI] [PMC free article] [PubMed] [Google Scholar]
- 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]
- White D. A., Lennarz W. J., Schnaitman C. A. Distribution of lipids in the wall and cytoplasmic membrane subfractions of the cell envelope of Escherichia coli. J Bacteriol. 1972 Feb;109(2):686–690. doi: 10.1128/jb.109.2.686-690.1972. [DOI] [PMC free article] [PubMed] [Google Scholar]