An N-linked high-mannose type oligosaccharide, expressed at the major outer membrane protein of Chlamydia trachomatis, mediates attachment and infectivity of the microorganism to HeLa cells.
The structure of the carbohydrate of the 40-kD major outer membrane component of Chlamydia trachomatis and its role in defining infectivity of the organism were investigated. The oligosaccharides were released from the glycoprotein by N-glycanase digestion, coupled to a 2-aminopyridyl residue, and subjected to two-dimensional sugar mapping technique. The major fractions consisted of "high-mannose type" oligosaccharides containing 8-9 mannose residues. Bi- and tri-antennary "complex type" oligosaccharides having terminal galactose were detected as minor components. These oligosaccharides were N-linked and contained no sialic acid. This structural profile is consistent with our previous characterization based on lectin-binding and glycosidase digestion. Functional specificity of identified chlamydial oligosaccharides was analyzed using glycopeptides fractionated from ovalbumin and structurally defined oligosaccharides from other sources. The glycopeptide fraction having high-mannose type oligosaccharide, as compared to those having complex or hybrid-type, showed a stronger inhibitory effect on attachment and infectivity of chlamydial organisms to HeLa cells. Among high-mannose type oligosaccharides, the strongest inhibition was observed with mannose 8 as compared with mannose 6, 7, or 9. These results indicate that a specific high-mannose type oligosaccharide linked to the major outer membrane protein of C. trachomatis mediates attachment and infectivity of the organism to HeLa cells.
Kuo
C
C
Department of Pathobiology, University of Washington, Seattle 98195, USA. cckuo@u.washington.edu
Takahashi
N
N
Swanson
A F
AF
Ozeki
Y
Y
Hakomori
S
S
D000581:Amidohydrolases / Q000378:metabolism; D001422:Bacterial Adhesion / Q000187:drug effects; D001425:Bacterial Outer Membrane Proteins / Q000737:chemistry* / Q000494:pharmacology; D002236:Carbohydrate Conformation; D002240:Carbohydrate Sequence; D002692:Chlamydia trachomatis / Q000737:chemistry*; D002845:Chromatography; D002846:Chromatography, Affinity; D006020:Glycopeptides / Q000494:pharmacology; D006367:HeLa Cells; D006801:Humans; D008362:Mannosides / Q000494:pharmacology; D008969:Molecular Sequence Data; D015394:Molecular Structure; D010047:Ovalbumin / Q000737:chemistry; D043524:Peptide-N4-(N-acetyl-beta-glucosaminyl) Asparagine Amidase; D011135:Polysaccharides, Bacterial / Q000737:chemistry* / Q000494:pharmacology; D018272:Porins
D001425:Bacterial Outer Membrane Proteins; D006020:Glycopeptides; D008362:Mannosides; D011135:Polysaccharides, Bacterial; D018272:Porins; C071677:omp1 protein, Chlamydia trachomatis; D010047:Ovalbumin; D000581:Amidohydrolases; D043524:Peptide-N4-(N-acetyl-beta-glucosaminyl) Asparagine Amidase