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. 1985 Sep;163(3):1080–1086. doi: 10.1128/jb.163.3.1080-1086.1985

Analysis of outer membrane proteins which are associated with growth of Bacteroides thetaiotaomicron on chondroitin sulfate.

S F Kotarski, J Linz, D M Braun, A A Salyers
PMCID: PMC219241  PMID: 2411713

Abstract

By analyzing outer membrane proteins of Bacteroides thetaiotaomicron on two-dimensional polyacrylamide gels, we were able to identify 10 protein spots that were associated with growth on chondroitin sulfate but not with growth on glucuronic acid or other monosaccharides. These proteins were distinct from the outer membrane polypeptides that were associated with growth on two other negatively charged polysaccharides, polygalacturonic acid and heparin. Of the 10 protein spots that were associated with growth on chondroitin sulfate, 4 could be detected on immunoblots with antiserum that had been raised against outer membranes from bacteria grown on chondroitin sulfate and then cross-adsorbed with membranes from bacteria grown on glucose. Synthesis of these four proteins appeared to be regulated coordinately with synthesis of the two enzymes that degrade chondroitin sulfate, chondroitin lyase I and II. Although one of the four proteins (Mr 110,000) was similar in molecular weight to the chondroitin lyases, the cross-adsorbed antiserum which detected this outer membrane protein did not cross-react with either of these two enzymes.

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Selected References

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  1. Ames G. F., Nikaido K. Two-dimensional gel electrophoresis of membrane proteins. Biochemistry. 1976 Feb 10;15(3):616–623. doi: 10.1021/bi00648a026. [DOI] [PubMed] [Google Scholar]
  2. Erickson P. F., Minier L. N., Lasher R. S. Quantitative electrophoretic transfer of polypeptides from SDS polyacrylamide gels to nitrocellulose sheets: a method for their re-use in immunoautoradiographic detection of antigens. J Immunol Methods. 1982 Jun 11;51(2):241–249. doi: 10.1016/0022-1759(82)90263-0. [DOI] [PubMed] [Google Scholar]
  3. Eschenbruch M., Bürk R. R. Experimentally improved reliability of ultrasensitive silver staining of protein in polyacrylamide gels. Anal Biochem. 1982 Sep 1;125(1):96–99. doi: 10.1016/0003-2697(82)90387-6. [DOI] [PubMed] [Google Scholar]
  4. Kotarski S. F., Salyers A. A. Isolation and characterization of outer membranes of Bacteroides thetaiotaomicron grown on different carbohydrates. J Bacteriol. 1984 Apr;158(1):102–109. doi: 10.1128/jb.158.1.102-109.1984. [DOI] [PMC free article] [PubMed] [Google Scholar]
  5. Kranz R. G., Gennis R. B. A quantitative radioimmunological screening method for specific gene products. Anal Biochem. 1982 Dec;127(2):247–257. doi: 10.1016/0003-2697(82)90169-5. [DOI] [PubMed] [Google Scholar]
  6. Laemmli U. K., Favre M. Maturation of the head of bacteriophage T4. I. DNA packaging events. J Mol Biol. 1973 Nov 15;80(4):575–599. doi: 10.1016/0022-2836(73)90198-8. [DOI] [PubMed] [Google Scholar]
  7. Linn S., Chan T., Lipeski L., Salyers A. A. Isolation and characterization of two chondroitin lyases from Bacteroides thetaiotaomicron. J Bacteriol. 1983 Nov;156(2):859–866. doi: 10.1128/jb.156.2.859-866.1983. [DOI] [PMC free article] [PubMed] [Google Scholar]
  8. Lopatin D. E., Voss E. W., Jr Anti-lysergyl antibody: measurement of binding parameters in IgG fractions. Immunochemistry. 1974 Jun;11(6):285–293. doi: 10.1016/0019-2791(74)90364-4. [DOI] [PubMed] [Google Scholar]
  9. Markwell M. A., Haas S. M., Bieber L. L., Tolbert N. E. A modification of the Lowry procedure to simplify protein determination in membrane and lipoprotein samples. Anal Biochem. 1978 Jun 15;87(1):206–210. doi: 10.1016/0003-2697(78)90586-9. [DOI] [PubMed] [Google Scholar]
  10. O'Farrell P. H. High resolution two-dimensional electrophoresis of proteins. J Biol Chem. 1975 May 25;250(10):4007–4021. [PMC free article] [PubMed] [Google Scholar]
  11. O'Farrell P. Z., Goodman H. M., O'Farrell P. H. High resolution two-dimensional electrophoresis of basic as well as acidic proteins. Cell. 1977 Dec;12(4):1133–1141. doi: 10.1016/0092-8674(77)90176-3. [DOI] [PubMed] [Google Scholar]
  12. Salyers A. A., Kotarski S. F. Induction of chondroitin sulfate lyase activity in Bacteroides thetaiotaomicron. J Bacteriol. 1980 Aug;143(2):781–788. doi: 10.1128/jb.143.2.781-788.1980. [DOI] [PMC free article] [PubMed] [Google Scholar]
  13. Salyers A. A., O'Brien M. Cellular location of enzymes involved in chondroitin sulfate breakdown by Bacteroides thetaiotaomicron. J Bacteriol. 1980 Aug;143(2):772–780. doi: 10.1128/jb.143.2.772-780.1980. [DOI] [PMC free article] [PubMed] [Google Scholar]
  14. Varel V. H., Bryant M. P. Nutritional features of Bacteroides fragilis subsp. fragilis. Appl Microbiol. 1974 Aug;28(2):251–257. doi: 10.1128/am.28.2.251-257.1974. [DOI] [PMC free article] [PubMed] [Google Scholar]

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