A lectin recognizes differential arrangements of O-glycans on mucin repeats

Interaction of Vicia villosa agglutinin-B4 (VVA-B4) to glycopeptides with O-linked GalNAc residues was investigated by surface plasmon resonance. The affinity was shown to be influenced by the arrangement of O-glycosylation sites on a peptide, PTTTPITTTTK, representing the tandem repeat of MUC2. The association rate constant was relatively high with a particular category of GalNAc-peptides in which more than three amino acid residues were placed between GalNAc-Thr residues. PTT ∗T ∗PITT ∗T ∗TK (T ∗ indicates GalNAc-Thr) had the highest association rate constant among the glycopeptides tested. The dissociation rate constant was low in the peptides containing consecutive GalNAc residues and PT ∗TTPIT ∗T ∗T ∗TK was the lowest of the glycopeptides tested. Dissociation constant ( K D), calculated as k d/ k a was the lowest with PTT ∗T ∗PITT ∗T ∗TK. Therefore, the arrangement but not the quantity of GalNAc residues apparently determines the affinity between VVA-B4 and peptides with attached GalNAc residues.