Effect of O-acetylation of O antigen of Escherichia coli lipopolysaccharide on the nonspecific barrier function of the outer membrane
- 118 Downloads
Comparison of the methods for determination of permeability of the outer membrane of Escherichia coli strain 4s and its mutants was carried out. The studied isogenic strains E. coli 4s were obtained by selection of spontaneous mutants according to their sensitivity to bacteriophages recognizing the surface O antigen of the outer membrane lipopolysaccharide as a primary receptor. The variants differed in the presence and (de)acetylation of the lipopolysaccharide O antigen. A peptide antibiotic polymyxin, plasmid DNA, and lysozyme were used as probes. The role of acetylation of the O antigen of the lipopolysaccaride of E. coli outer membrane in modification of its permeability (correlating with bacteriophage sensitivity of the cells) was confirmed. Kinetic analysis using lysozyme was shown to be the optimal method for determination of the barrier function of E. coli outer membrane.
KeywordsEscherichia coli spontaneous mutants bacteriophage sensitivity lipopolysaccharide O antigen polymyxin lysozyme outer membrane permeability O antigen acetylation
Unable to display preview. Download preview PDF.
- Golomidova, A.K., Kulikov, E.E., Prokhorov, N.S., Guerrero-Ferreira, R., Knirel, Y.A., Kostryukova, E.S., Tarasyan, K.K., and Letarov A.V, Branched lateral tail fiber organization in T5-like bacteriophages DT57C and DT571/2 is revealed by genetic and functional analysis, Viruses, 2016, vol. 8, no. 1. pii: E26. doi 10.3390/v8010026CrossRefPubMedGoogle Scholar
- Hitchcock, P.J, Analyses of gonococcal lipopolysaccharide in whole-cell lysates by sodium dodecyl sulfate-polyacrylamide gel electrophoresis: stable association of lipopolysaccharide with the major outer membrane protein (protein I) of Neisseria gonorrhoeae, Infect. Immun., 1984, vol. 46, pp. 202–212.PubMedPubMedCentralGoogle Scholar
- Knirel, Y.A., Prokhorov, N.S., Shashkov, A.S., Ovchinnikova, O.G., Zdorovenko, E.L., Liu, B., Kostryukova, E.S., Larin, A.K., Golomidova, A.K., and Letarov, A.V, Variations in O-antigen biosynthesis and O-acetylation associated with altered phage sensitivity in Escherichia coli 4s, J. Bacteriol., 2015, vol. 197, pp. 905–912.CrossRefPubMedPubMedCentralGoogle Scholar
- Kulikov, E., Kropinski, A.M., Golomidova, A., Lingohr, E., Govorun, V., Serebryakova, M., Prokhorov, N., Letarova, M., Manykin, A., Strotskaya, A., and Letarov, A, Isolation and characterization of a novel indigenous intestinal N4-related coliphage vB_EcoP_G7C, Virology, 2012, vol. 426, pp. 93–99.CrossRefPubMedGoogle Scholar
- Singh, A. and Reithmeier, R, Leakage of periplasmic enzymes from cells of heptose-deficient mutants of Escherichia coli, associated with alterations in the protein component of the outer membrane, J. Gen. Appl. Microbiol., 1975, vol. 21, pp. 109–118.Google Scholar
- Tsai, C.M. and Frasch, C.E., A sensitive silver stain for detecting lipopolysaccharides in polyacrylamide gels, Anal. Biochem., 1982, vol. 119, pp. 115–119.Google Scholar
- Tu, Q., Yin, J., Fu, J., Herrmann, J., Li, Y., Yin, Y., Stewart, A.F., Muller, R., and Zhang, Y., Room temperature electrocompetent bacterial cells improve DNA transformation and recombineering efficiency, Sci. Rep., 2016, vol. 6, P. 24648. doi 10.1038/srep24648CrossRefPubMedPubMedCentralGoogle Scholar