Structural requirements for the binding of oligosaccharides and glycopeptides to immobilized wheat germ agglutinin

Abstract

The structural requirements for the interaction of asparagine-linked oligosaccharide moieties of glycoproteins with wheat germ agglutinin (WGA) were investigated by using affinity chromatography on a WGA-Sepharose column. So-called hybrid-type glycopeptides obtained from ovalbumin [Yamashita, K., Tachibana, Y., & Kobata, A. (1978) J. Biol. Chem. 253, 3862--3869] were found to have high affinity for WGA--Sepharose, whereas high mannose-type and complex-type glycopeptides were shown to have low affinity. The elution profiles of various glycopeptides modified by glycosidase treatment, Smith periodate degradation, acetolysis, and hydrazinolysis showed that the GlcNAcbeta 1--4Manbeta 1--4GlcNAc beta 1--4GlcNAc-Asn structure was essential for the binding of glycopeptides to a WGA-Sepharose column. Thus, it was revealed that both the N,N'-diacetylchitobiose moiety and the beta-N-acetylglucosaminyl residue linked to C-4 of the beta-linked mannose residue contributed to the interaction of the glycopeptide with WGA-Sepharose. The substitution at C-6 of the innermost beta-N-acetylglucosaminyl residue by an alpha-fucosyl residue or at C-6 of the beta-linked mannose residue by another mannose residue in the above structure reduced the affinity of glycopeptides for the column.

Glycosylation changes in different developmental stages of Trichinella

“…On mammalian cell surfaces, GlcNAc residues have been found in continuous sequences only in Nlinked oligosaccharides, where they form the core that is attached to the asparagine. The binding of such components is weak in the absence of the bisecting GlcNAc residue that is present in the hybrid type N-bound oligosaccharides (Yamamoto et al 1981). GlcNAc is highly represented in T. spiralis N-glycans, where it is either in terminal position or it is implicated in the so called "lacdiNAC" antennae.…”

The Coactivator of Transcription CREB-binding Protein Interacts Preferentially with the Glycosylated Form of Stat5

“…1B). WGA lectin has a specific affinity for terminal GlcNAc residues and allows the separation of Stat5a into glycosylated and non-glycosylated fractions (37).…”

Isolation and Characterization of a Diagnostic Polysaccharide Antigen From Larval Echinococcus Multilocularis

“…These results suggest that C-antigen contains at least galactose and N-acetyl glucosamine residues. RCA120 is specific to galactose or lactose residues (11), PNA to galactose residues (12) and WGA to N-acetyl glucosamine or sialic acid residues (13).…”