Use of monoclonal anti-light subunit antibodies to study the structure and function of the Entamoeba histolytica Gal/GalNAc adherence lectin.

McCoy JJ, Weaver AM, Petri WA
Glycoconj J. 1994 11 (5): 432-6

PMID: 7696848

Adherence of Entamoeba histolytica trophozoites to host cells is mediated by a galactose (Gal) and N-acetylgalactosamine (GalNAc)-specific surface lectin. The lectin is a heterodimeric protein composed of heavy (170 kDa) and light (35-31 kDa) subunits linked by disulfide bonds. Polyclonal and monoclonal antibodies (mAb) raised against a light subunit-glutathione-S-transferase fusion protein were used to probe its structure and function. Four light subunit-specific mAb were produced which recognized distinct epitopes on five different light subunit isoforms. Immunoblots with these mAb demonstrated co-migration of light and heavy subunits when nonreduced trophozoite proteins were analysed by SDS-PAGE, indicating that the subunits do not exist free of the heterodimer in significant quantities. While anti-heavy subunit antibodies had previously been shown to alter adherence, anti-light subunit antibodies did not, suggesting that the heavy subunit contains the carbohydrate recognition domain.

MeSH Terms (16)

Acetylgalactosamine Amino Acid Sequence Animals Antibodies, Monoclonal Blotting, Western CHO Cells Chromatography, Affinity Cricetinae Entamoeba histolytica Galactose Lectins Membrane Glycoproteins Molecular Sequence Data Protein Binding Protozoan Proteins Structure-Activity Relationship

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