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. 1991 Jun;59(6):2120–2125. doi: 10.1128/iai.59.6.2120-2125.1991

Evidence that the major outer membrane protein of Chlamydia trachomatis is glycosylated.

A F Swanson 1, C C Kuo 1
PMCID: PMC257975  PMID: 1645328

Abstract

The major outer membrane protein (MOMP) of Chlamydia trachomatis was determined to be a glycoprotein on the basis of susceptibility to glycosidase digestion and the presence of carbohydrate by staining and radiolabeling. The MOMP of the serovar L2 organisms was isolated by electroelution from the protein band excised from the gel after sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE). The incubation of MOMP with N-glycosidase F, an endoglycosidase that cleaves the N-glycan, and periodate resulted in two new molecular weight species. While MOMP treated with N-glycosidase F showed a lower-molecular-weight mobility, the periodate-treated MOMP increased in molecular weight. Both treatments abolished the ability of the MOMP to bind to HeLa cell components. In the immunoblot, the reactivity to the monoclonal antibody specific against the C. trachomatis species was preserved. The endoglycosidase specific to O-linked glycan, endo-alpha-N-acetylgalactosaminidase, had no visible effect on the isolated MOMP. Carbohydrate was detected in the MOMP by p-phenylenediamine staining of the protein band in the gel following SDS-PAGE. Autoradiograms of proteins of chlamydial organisms metabolically labeled with [3H]galactose or [3H]glucosamine and separated by SDS-PAGE revealed the MOMP band. The isolated MOMP was shown to bind specifically to concanavalin A, wheat germ agglutinin, and Dolichos biflorus agglutinin in the lectin binding assay. No binding was observed with Ulex europaeus agglutinin I, soybean agglutinin, or Ricinus communis agglutinin.

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Selected References

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

  1. Bose S. K., Smith G. B., Paul R. G. Influence of lectins, hexoses, and neuraminidase on the association of purified elementary bodies of Chlamydia trachomatis UW-31 with HeLa cells. Infect Immun. 1983 Jun;40(3):1060–1067. doi: 10.1128/iai.40.3.1060-1067.1983. [DOI] [PMC free article] [PubMed] [Google Scholar]
  2. Caldwell H. D., Judd R. C. Structural analysis of chlamydial major outer membrane proteins. Infect Immun. 1982 Dec;38(3):960–968. doi: 10.1128/iai.38.3.960-968.1982. [DOI] [PMC free article] [PubMed] [Google Scholar]
  3. Caldwell H. D., Kromhout J., Schachter J. Purification and partial characterization of the major outer membrane protein of Chlamydia trachomatis. Infect Immun. 1981 Mar;31(3):1161–1176. doi: 10.1128/iai.31.3.1161-1176.1981. [DOI] [PMC free article] [PubMed] [Google Scholar]
  4. Chi E. Y., Kuo C. C., Grayston J. T. Unique ultrastructure in the elementary body of Chlamydia sp. strain TWAR. J Bacteriol. 1987 Aug;169(8):3757–3763. doi: 10.1128/jb.169.8.3757-3763.1987. [DOI] [PMC free article] [PubMed] [Google Scholar]
  5. Elbein A. D. Inhibitors of the biosynthesis and processing of N-linked oligosaccharide chains. Annu Rev Biochem. 1987;56:497–534. doi: 10.1146/annurev.bi.56.070187.002433. [DOI] [PubMed] [Google Scholar]
  6. Farach-Carson M. C., Carson D. D. Extraction and isolation of glycoproteins and proteoglycans. Biotechniques. 1989 May;7(5):482–493. [PubMed] [Google Scholar]
  7. Hackstadt T. Identification and properties of chlamydial polypeptides that bind eucaryotic cell surface components. J Bacteriol. 1986 Jan;165(1):13–20. doi: 10.1128/jb.165.1.13-20.1986. [DOI] [PMC free article] [PubMed] [Google Scholar]
  8. Kuo C. C., Wang S. P., Grayston J. T. Effect of polycations, polyanions and neuraminidase on the infectivity of trachoma-inclusin conjunctivitis and lymphogranuloma venereum organisms HeLa cells: sialic acid residues as possible receptors for trachoma-inclusion conjunction. Infect Immun. 1973 Jul;8(1):74–79. doi: 10.1128/iai.8.1.74-79.1973. [DOI] [PMC free article] [PubMed] [Google Scholar]
  9. Köttgen E., Hell B., Müller C., Tauber R. Demonstration of glycosylation variants of human fibrinogen, using the new technique of glycoprotein lectin immunosorbent assay (GLIA). Biol Chem Hoppe Seyler. 1988 Oct;369(10):1157–1166. doi: 10.1515/bchm3.1988.369.2.1157. [DOI] [PubMed] [Google Scholar]
  10. Laemmli U. K. Cleavage of structural proteins during the assembly of the head of bacteriophage T4. Nature. 1970 Aug 15;227(5259):680–685. doi: 10.1038/227680a0. [DOI] [PubMed] [Google Scholar]
  11. Lechner J., Wieland F. Structure and biosynthesis of prokaryotic glycoproteins. Annu Rev Biochem. 1989;58:173–194. doi: 10.1146/annurev.bi.58.070189.001133. [DOI] [PubMed] [Google Scholar]
  12. Levy N. J. Wheat germ agglutinin blockage of chlamydial attachment sites: antagonism by N-acetyl-D-glucosamine. Infect Immun. 1979 Sep;25(3):946–953. doi: 10.1128/iai.25.3.946-953.1979. [DOI] [PMC free article] [PubMed] [Google Scholar]
  13. Newhall W. J., Jones R. B. Disulfide-linked oligomers of the major outer membrane protein of chlamydiae. J Bacteriol. 1983 May;154(2):998–1001. doi: 10.1128/jb.154.2.998-1001.1983. [DOI] [PMC free article] [PubMed] [Google Scholar]
  14. Peterson E. M., de la Maza L. M. Chlamydia parasitism: ultrastructural characterization of the interaction between the chlamydial cell envelope and the host cell. J Bacteriol. 1988 Mar;170(3):1389–1392. doi: 10.1128/jb.170.3.1389-1392.1988. [DOI] [PMC free article] [PubMed] [Google Scholar]
  15. Segrest J. P., Jackson R. L., Andrews E. P., Marchesi V. T. Human erythrocyte membrane glycoprotein: a re-evaluation of the molecular weight as determined by SDS polyacrylamide gel electrophoresis. Biochem Biophys Res Commun. 1971 Jul 16;44(2):390–395. doi: 10.1016/0006-291x(71)90612-7. [DOI] [PubMed] [Google Scholar]
  16. Stephens R. S., Sanchez-Pescador R., Wagar E. A., Inouye C., Urdea M. S. Diversity of Chlamydia trachomatis major outer membrane protein genes. J Bacteriol. 1987 Sep;169(9):3879–3885. doi: 10.1128/jb.169.9.3879-3885.1987. [DOI] [PMC free article] [PubMed] [Google Scholar]
  17. Stephens R. S., Tam M. R., Kuo C. C., Nowinski R. C. Monoclonal antibodies to Chlamydia trachomatis: antibody specificities and antigen characterization. J Immunol. 1982 Mar;128(3):1083–1089. [PubMed] [Google Scholar]
  18. Su H., Watkins N. G., Zhang Y. X., Caldwell H. D. Chlamydia trachomatis-host cell interactions: role of the chlamydial major outer membrane protein as an adhesin. Infect Immun. 1990 Apr;58(4):1017–1025. doi: 10.1128/iai.58.4.1017-1025.1990. [DOI] [PMC free article] [PubMed] [Google Scholar]
  19. Swanson A. F., Kuo C. C. Evidence that the major outer membrane protein of Chlamydia trachomatis is glycosylated. Infect Immun. 1991 Jun;59(6):2120–2125. doi: 10.1128/iai.59.6.2120-2125.1991. [DOI] [PMC free article] [PubMed] [Google Scholar]
  20. Swanson A. F., Kuo C. C. Identification of lectin-binding proteins in Chlamydia species. Infect Immun. 1990 Feb;58(2):502–507. doi: 10.1128/iai.58.2.502-507.1990. [DOI] [PMC free article] [PubMed] [Google Scholar]
  21. Towbin H., Staehelin T., Gordon J. Electrophoretic transfer of proteins from polyacrylamide gels to nitrocellulose sheets: procedure and some applications. Proc Natl Acad Sci U S A. 1979 Sep;76(9):4350–4354. doi: 10.1073/pnas.76.9.4350. [DOI] [PMC free article] [PubMed] [Google Scholar]
  22. Twining S. S. Fluorescein isothiocyanate-labeled casein assay for proteolytic enzymes. Anal Biochem. 1984 Nov 15;143(1):30–34. doi: 10.1016/0003-2697(84)90553-0. [DOI] [PubMed] [Google Scholar]
  23. URIEL J. Detection des activités catalasiques et peroxydasiques de l'hémoglobine après électrophorèse en gélose. Bull Soc Chim Biol (Paris) 1958;40(1):277–280. [PubMed] [Google Scholar]
  24. Wenman W. M., Meuser R. U. Chlamydia trachomatis elementary bodies possess proteins which bind to eucaryotic cell membranes. J Bacteriol. 1986 Feb;165(2):602–607. doi: 10.1128/jb.165.2.602-607.1986. [DOI] [PMC free article] [PubMed] [Google Scholar]
  25. Zhang Y. X., Morrison S. G., Caldwell H. D., Baehr W. Cloning and sequence analysis of the major outer membrane protein genes of two Chlamydia psittaci strains. Infect Immun. 1989 May;57(5):1621–1625. doi: 10.1128/iai.57.5.1621-1625.1989. [DOI] [PMC free article] [PubMed] [Google Scholar]

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