Studies on the binding site of the galactose-specific agglutinin PA-IL from Pseudomonas aeruginosa

• Published in: Glycobiology (Oxford) Vol. 8; no. 1; pp. 7 - 16
• Main Authors: Chen, Chie-Pein, Song, Shuh-Chyung, Gilboa-Garber, Nechama, Chang, Kenneth S.S., Wu, Albert M.
• Format: Journal Article
• Language: English
• Published: England Oxford University Press 01-01-1998
• Subjects: ABO Blood-Group System - metabolism; Adhesins, Bacterial - chemistry; Adhesins, Bacterial - metabolism; Animals; binding properties; Binding Sites; Carbohydrate Conformation; Carbohydrate Sequence; combining sites; glycoproteins; Glycoproteins - chemistry; Glycoproteins - metabolism; Humans; In Vitro Techniques; lectin; Lectins - chemistry; Lectins - metabolism; Molecular Sequence Data; Polysaccharides - chemistry; Polysaccharides - metabolism; Pseudomonas aeruginosa; Pseudomonas aeruginosa - metabolism
• ISSN: 0959-6658; 1460-2423
• DOI: 10.1093/glycob/8.1.7

The binding properties of Pseudomonas aeruginosa agglutinin-I (PA-IL) with glycoproteins (gps) and polysaccharides were studied by both the biotin/avidin-mediated microtiter plate lectin-binding assay and the inhibition of agglutinin-glycan interaction with sugar ligands. Among 36 glycans tested for binding, PA-IL reacted best with two glycoproteins containing Galα1→4Gal determinants and a human blood group ABO precursor equivalent gp, but this lectin reacted weakly or not at all with A and H active gps or sialylated gps. Among the mammalian disaccharides tested by the inhibition assay, the human blood group Pk active Galα1→4Gal, was the best. It was 7.4-fold less active than melibiose (Galα1→6Glc). PA-IL has a preference for the α-anomer in decreasing order as follows: Galα1→6 > Galα1→4 > Galα1→3. Of the monosaccharides studied, the phenylβ derivatives of Gal were much better inhibitors than the methylβ derivative, while only an insignificant difference was found between the Galα anomer of methyl- and p-NO2-phenyl derivatives. From these results, it can be concluded that the combining size of the agglutinin is as large as a disaccharide of the α-anomer of Gal at nonreducing end and most complementary to Galα1→6Glc. As for the combining site of PA-IL toward the β-anomer, the size is assumed to be less than that of Gal; carbon-6 in the pyranose form is essential, and hydrophobic interaction is important for binding.