Interactions of the fucose-specific Pseudomonas aeruginosa lectin, PA-IIL, with mammalian glycoconjugates bearing polyvalent Lewis a and  ABH blood group glycotopes

Pseudomonas aeruginosa Fuc > Man specific lectin, PA-IIL, is an important microbial agglutinin that might be involved in P. aeruginosa infections in humans. In order to delineate the structures of these lectin receptors, its detailed carbohydrate recognition profile was studied both by microtiter plate biotin/avidin-mediated enzyme–lectin–glycan binding assay (ELLSA) and by inhibition of the lectin–glycan interaction. Among 40 glycans tested for binding, PA-IIL reacted well with all human blood group ABH and Le a/ Le b active glycoproteins (gps), but weakly or not at all with their precursor gps and N-linked gps. Among the sugar ligands tested by the inhibition assay, the Le a pentasaccharide lacto- N-fucopentaose II ( LNFP II, Galβ1-3[Fucα1-4]GlcNAcβ1-3Galβ1-4Glc) was the most potent one, being 10 and 38 times more active than the Le x pentasaccharide ( LNFP III, Galβ1-4 [Fucα1-3]GlcNAcβ1-3Galβ1-4Glc) and sialyl Le x (Neu5Acα2-3Galβ1-4[Fucα1-3] GlcNAc), respectively. It was 120 times more active than Man, while Gal and GalNAc were inactive. The decreasing order of PA-IIL affinity for the oligosaccharides tested was: Le a pentaose ≥ sialyl Le a tetraose > methyl αFuc > Fuc and Fucα1-2Gal ( H disaccharide) > 2′-fucosyllactose ( H trisaccharide), Le x pentaose, Le b hexaose ( LNDFH I) and gluco-analogue of Le y tetraose ( LDFT) > H type I determinant ( LNFP I) > Le x trisaccharide (Galβ1-4[Fucα1-3]GlcNAc) > sialyl Le x trisaccharide >> Man >>> Gal, GalNAc, and Glc (inactive). The results presented here, in accordance with the crystal 3D structural data, imply that the combining site of PA-IIL is a small cavity-type best fitting Fucα1- with a specific shallow groove subsite for the remainder part of the Le a saccharides, and that polyvalent glycotopes enhance the reactivity. The Fuc > Man Ralstonia solanacearum lectin RSL, which resembles PA-IIL in sugar specificity, differs from it in it's better fit to the B and A followed by H oligosaccharides vs. Fuc, whereas, the second R. solanacearum lectin RS-IIL (the structural homologue of PA-IIL) binds Man > Fuc. These results provide a valuable information on PA-IIL interactions with mammalian glycoforms and the possible spectrum of attachment sites for the homing of this aggressive bacterium onto the target molecules. Such information might be useful for the antiadhesive therapy of P. aeruginosa infections.