title: Epitope expression and partial structural characterization of F62 lipooligosaccharide (LOS) of Neisseria gonorrhoeae: IgM monoclonal antibodies (3F11 and 1-1-M) recognize non-reducing termini of the LOS components.
abstract: F62 LOS of Neisseria gonorrhoeae consists of two components. The higher molecular weight (MW) component is recognized by monoclonal antibody (MAb) 1-1-M and the smaller MW component by MAb 3F11. Epitope expression of the two LOS components and their partial structures were investigated by treating the F62 LOS with several glycosidases and then monitoring their antigenicity with the two mouse IgM MAbs. The 1-1-M-defined LOS component was cleaved with both beta-N-acetylhexosaminidase and endo-beta-galactosidase, and each cleavage resulted in the loss of expression of the 1-1-M-defined epitope. The N-acetylhexosamine (HexNAc) released by the hexosaminidase was found to be GalNAc, and the smaller oligosaccharide released by the endo enzyme was identified to be a dimer GalNAc beta----Gal. In contrast, the MAb 3F11-defined LOS component was not digested by the endo galactosidase, but it was cleaved with alpha and beta-galactosidase, and expression of the MAb 3F11-defined LOS epitope expression of the MAb 3F11-defined LOS was abolished by the treatment with each of two exo enzymes. MAb 3F11 bound to the 1-1-M-defined LOS component resulting from the removal of the beta-GalNAc residue, and the resulting LOS was further cleaved with beta-galactosidase, but not with alpha-galactosidase. From these results, we conclude the following: (1) MAbs 1-1-M and 3F11 both recognize the non-reducing termini of the LOS components; (2) the 1-1-M-defined LOS component has the GalNAc beta----Gal beta 1----4-Glc (or GlcNAc) structure, and the GalNAc beta----Gal residue is involved in the MAb 1-1-M-defined epitope; (3) the MAb 3F11-defined LOS component may not have a Gal beta 1----4GlcNAc beta 1----4Gal beta 1----4Glc structure within the molecule. However, it has beta-Gal residue at its non-reducing terminus, and this residue is involved in the MAb 3F11-defined epitope; (4) the two LOS components share a similar antigenic structure, and the 3F11-defined epitope structure is present in the MAb 1-1-M-defined LOS component. Expression of this epitope within the 1-1-M-defined LOS molecule is blocked by the beta-GalNAc residue; however, the beta-GalNAc residue at the non-reducing end may be not the only structural difference between the two components.
authors.affiliation: Center for Immunochemistry, University of California, San Francisco.
authors.forename: R
authors.initials: R
authors.lastname: Yamasaki
authors.forename: W
authors.initials: W
authors.lastname: Nasholds
authors.forename: H
authors.initials: H
authors.lastname: Schneider
authors.forename: M A
authors.initials: MA
authors.lastname: Apicella
mesh_terms: D000118:Acetylglucosaminidase / Q000494:pharmacology; D000911:Antibodies, Monoclonal / Q000276:immunology*; D002855:Chromatography, Thin Layer; D004591:Electrophoresis, Polyacrylamide Gel; D000939:Epitopes / Q000096:biosynthesis*; D005786:Gene Expression Regulation / Q000187:drug effects; D006026:Glycoside Hydrolases; D015151:Immunoblotting; D007075:Immunoglobulin M / Q000235:genetics*; D008070:Lipopolysaccharides / Q000737:chemistry*; D009344:Neisseria gonorrhoeae / Q000235:genetics / Q000276:immunology*; D000519:alpha-Galactosidase / Q000494:pharmacology; D001616:beta-Galactosidase / Q000494:pharmacology
chemical_list: D000911:Antibodies, Monoclonal; D000939:Epitopes; C071915:F62 lipooligosaccharide; D007075:Immunoglobulin M; D008070:Lipopolysaccharides; D006026:Glycoside Hydrolases; C026093:keratan-sulfate endo-1,4-beta-galactosidase; D000519:alpha-Galactosidase; D001616:beta-Galactosidase; D000118:Acetylglucosaminidase