Multi-specificity of a Psathyrella velutina mushroom lectin: heparin/pectin binding occurs at a site different from the N-acetylglucosamine/N-acetylneuraminic acid-specific site.
An N-acetylglucosamine (GlcNAc)/N-acetylneuraminic acid-specific lectin from the fruiting body of Psathyrella velutina (PVL) is a useful probe for the detection and fractionation of specific carbohydrates. In this study, PVL was found to exhibit multispecificity to acidic polysaccharides and sulfatides. Purified PVL and a counterpart lectin to PVL in the mycelium interact with heparin neoproteoglycans, as detected by both membrane analysis and solid phase assay. The pH-dependencies of the binding to heparin and GlcNAc5-6 differ. The heparin binding of PVL is inhibited best by pectin, polygalacturonic acid, and highly sulfated polysaccharides, but not by GlcNAc, colominic acid, or other glycosaminoglycans. Sandwich affinity chromatography indicated that PVL can simultaneously interact with heparin- and GlcNAc-containing macromolecules. Extensive biotinylation was found to suppress the binding activity to heparin while the GlcNAc binding activity is retained. On the other hand, biotinyl PVL binds to sulfatide and the binding is not inhibited by GlcNAc, N-acetylneuraminic acid, or heparin. These results indicate that PVL is a multi-ligand adhesive lectin that can interact with various glycoconjugates. This multispecificity needs to be recognized when using PVL as a sugar-specific probe to avoid misleading information about the nature of glycoforms.
Ueda
H
H
Department of Advanced Biosciences, Graduate School of Humanities and Sciences, Kiryuu, Gunma, 376-0051, Japan.
Saitoh
T
T
Kojima
K
K
Ogawa
H
H
D000117:Acetylglucosamine / Q000378:metabolism*; D000363:Agaricales / Q000378:metabolism*; D001665:Binding Sites; D002846:Chromatography, Affinity; D002942:Circular Dichroism; D006493:Heparin / Q000378:metabolism*; D037102:Lectins / Q000378:metabolism*; D019158:N-Acetylneuraminic Acid / Q000378:metabolism*; D009994:Osmolar Concentration; D010368:Pectins / Q000378:metabolism*; D011485:Protein Binding; D013433:Sulfoglycosphingolipids / Q000378:metabolism
D037102:Lectins; D013433:Sulfoglycosphingolipids; D010368:Pectins; D006493:Heparin; D019158:N-Acetylneuraminic Acid; D000117:Acetylglucosamine