Delineation of the epitope recognized by an antibody specific for N- glycolylneuraminic acid-containing gangliosides

Ernesto Moreno, Boel Lanne, Ana María Vázquez, Ikuo Kawashima, Tadashi Tai, Luis Enrique Fernández, Karl Anders Karlsson, Jonas Ångström, Rolando Pérez

Research output: Contribution to journalArticlepeer-review

32 Scopus citations

Abstract

P3 is a mouse monoclonal antibody (mAb) that binds to several NeuGc- containing gangliosides. It also reacts with antigens expressed in human breast tumors (Vazquez et al. (1995) Hybridoma, 14, 551-556). In this work, the binding specificity of P3 has been characterized in more detail using a panel of glycolipids that included several disialylated gangliosides and several chemical derivatives of NeuGc-GM3. The carboxyl group and the nitrogen function of sialic acid were found to play important roles in the antibody binding, whereas the glycerol tail appears to be nonrelevant. Molecular modeling was used to analyze the binding data, including the finding that P3 selectively recognizes the internal NeuGc in GD3. For this purpose, conformational studies of GD3 were performed using molecular dynamics. It was concluded that sialic acid binds the P3 antibody through its upper face (the one on which the carboxyl group is exposed) and the C4-C5 side of the sugar ring, whereas none or very little contact between the galactose residue and the protein is evident. Conformational analysis of GD3 revealed that, despite the large flexibility of the NeuGcα8NeuGc linkage, the P3 binding epitope on the external sialic acid is not well exposed for any of the possible conformations this linkage can adopt, whereas the internal sialic acid presents the epitope in a proper way for several of these conformations. As a final result, a coherent picture of the epitope that fits the wide binding data was obtained.

Original languageEnglish
Pages (from-to)695-705
Number of pages11
JournalGlycobiology
Volume8
Issue number7
DOIs
StatePublished - Jul 1998
Externally publishedYes

Keywords

  • Epitope
  • Ganglioside
  • Molecular modeling
  • Monoclonal antibody
  • NeuGc

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