Binding of the glycan of the major outer membrane protein of Chlamydia trachomatis to HeLa cells.
Recent studies have shown that the major outer membrane protein (MOMP) of Chlamydia trachomatis is glycosylated. The glycan of the MOMP of C. trachomatis serovar L2 was separated from the glycoprotein with N-glycanase, reduced with tritiated NaBH4, and tested for its ability to interact with HeLa cells. The [3H]glycan was shown to attach readily to HeLa cells at 25 or 37 degrees C. This process was slower at 4 degrees C. Competition for possibly similar receptor sites on HeLa cells between the glycan and a sugar, an aminosaccharide, or elementary bodies (EBs) was then studied. D-Galactose, D-mannose, or N-acetylglucosamine was shown to reduce the attachment of the glycan to HeLa cells at concentrations of 0.1 to 0.5 M. Sedoheptulose, D-fructose, or sialic acid did not inhibit the binding of glycan to HeLa cells. The presence of at least 100 native or UV-inactivated EBs per HeLa cell interfered with the glycan's ability to bind to HeLa cells. Heat-inactivated EBs did not compete with the glycan for binding. In the reverse situation, nonradiolabeled glycan prevented the EBs from infecting and forming inclusions in HeLa cells. Incubation of [3H]glycan with rabbit immune serum prepared against antigens of whole EB and the MOMP inhibited attachment. In contrast, incubation of glycan with mouse monoclonal antibodies against the protein portion of the MOMP or the chlamydial lipopolysaccharide did not inhibit attachment. These results suggest that the glycan portion of the MOMP is involved in the attachment process of C. trachomatis organisms to HeLa cells.
Swanson
A F
AF
Department of Pathobiology, University of Washington, Seattle 98195.
Kuo
C C
CC
D001422:Bacterial Adhesion; D001425:Bacterial Outer Membrane Proteins / Q000378:metabolism*; D001667:Binding, Competitive; D002692:Chlamydia trachomatis / Q000254:growth & development / Q000378:metabolism*; D006367:HeLa Cells; D006801:Humans; D066298:In Vitro Techniques; D011134:Polysaccharides / Q000378:metabolism*
D001425:Bacterial Outer Membrane Proteins; D011134:Polysaccharides