Agaricus bisporus lectin binds mainly O-glycans but also N-glycans of human IgA subclasses

The primary interaction between purified Agaricus bisporus lectin (ABL) and human IgA subclasses was studied by ABL-affinity chromatography, dot blot assay and competitive enzyme-lectin assay, considering that ABL could be an alternative tool for detection of IgA1 O-glycans. Both secretory IgA subcl...

Full description

Saved in:
Bibliographic Details
Published in:Glycoconjugate journal 1997-04, Vol.14 (3), p.313-319
Main Authors: Irazoqui, F J, Zalazar, F E, Nores, G A, Vides, M A
Format: Article
Language:English
Subjects:
Agaricus bisporus
Bacteria
Chromatography
Chromatography, Affinity
Disaccharides
Enzyme immunoassay
Enzymes
Erythrina
Female
glycoconjugates
Glycosylation
Hemagglutination Inhibition Tests
horseradish peroxidase
Humans
Hydrolysis
Immunodiffusion
Immunoglobulin A
Immunoglobulin A - chemistry
Immunoglobulin A - classification
Immunoglobulin A - isolation & purification
Immunoglobulin A, Secretory - chemistry
Immunoglobulin A, Secretory - isolation & purification
Immunoglobulin G
Lectins
Milk, Human - immunology
Mushrooms
N-glycans
N-glycosidase
Optical density
Peptides
Phosphate
Polysaccharides - analysis
Standard deviation
Citations: Items that cite this one
Online Access:Get full text
Tags: Add Tag
No Tags, Be the first to tag this record!
Description
Summary:The primary interaction between purified Agaricus bisporus lectin (ABL) and human IgA subclasses was studied by ABL-affinity chromatography, dot blot assay and competitive enzyme-lectin assay, considering that ABL could be an alternative tool for detection of IgA1 O-glycans. Both secretory IgA subclasses bound to ABL-Sepharose and the IgA2 subclass (which contains only N-glycans) was recovered with a high degree of purity when NH4OH was used as eluent. ABL-Ig interaction was also observed by dot blot assays using ABL-peroxidase against monoclonal IgA1 k Pan, IgA2m(1)k Gir, IgA2m(2)k Bel, secretory IgA2 and normal IgG (also contains only N-glycans). When these immunoglobulins were enzymatically treated with peptide N-glycosidase F (N-glycan hydrolysis), the ABL-IgA2 and -IgG interaction did not occur while IgA1 maintained a high degree of interaction with ABL. Also, the ABL-IgA interaction was observed by competitive enzyme-lectin assay, and when IgA1 subclass was treated with endo-alpha-N-acetylgalactosaminidase for O-glycans hydrolysis, the reactivity with ABL was very low. We conclude that the complementary use of ABL and peptide N-glycosidase F could be a useful tool to assess the O-glycosylation state of human IgA1 subclass, which is of relevant importance in the effector functions of immunoglobulins.
ISSN:0282-0080
1573-4986
DOI:10.1023/a:1018566527271