Carbohydrate binding specificity of the Tn-antigen binding lectin from Vicia villosa seeds (VVLB4).
2-Dansylamino-2-deoxy-D-galactose (GalNDns) is a useful fluorescent probe to study the interaction of non-fluorescent sugars with the B4 lectin from Vicia villosa seeds (VVLB4). Binding of the lectin to GalNDns leads to a 5.2-fold increase in Dansyl fluorescence with a concomitant 10 nm blue shift in its emission maximum. The strong binding of GalNDns (Ka = 7.33 x 10(4) M-1 at 20 degrees C) is due to a favourable entropic contribution to the association process. Among the other sugars studied, GalNAc alpha 1-O-Ser followed by Me alpha GalNAc are the best ligands. 2-Deoxygalactose, galactosamine and galactose are 2013, 469 and 130 times weaker ligands, respectively, as compared to GalNAc, whereas GalNDns is about 2.44 times more potent than GalNAc, indicating that substitutions at the C-2 position of GalNAc have a considerable influence on the binding affinities. Equatorial orientation of the hydroxyl group at C-3 and axial orientation at C-4 as in galactose are important for the interaction with VVLB4. The C-6 hydroxyl group is not indispensable. The binding site of the lectin is directed exclusively towards monosaccharides alone. Interestingly enough, despite its preference for Me alpha GalNAc over Me beta GalNAc, in oligosaccharides, the lectin prefers terminal beta-linked GalNAc as compared to the alpha-linked one.
Puri
K D
KD
Molecular Biophysics Unit, Indian Institute of Science, Bangalore.
Gopalakrishnan
B
B
Surolia
A
A
D000941:Antigens / Q000378:metabolism; D050260:Carbohydrate Metabolism; D002240:Carbohydrate Sequence; D003619:Dansyl Compounds / Q000378:metabolism*; D005456:Fluorescent Dyes / Q000378:metabolism*; D005688:Galactosamine / Q000031:analogs & derivatives* / Q000378:metabolism; D037102:Lectins / Q000378:metabolism*; D008969:Molecular Sequence Data; D037121:Plant Lectins; D010944:Plants / Q000378:metabolism; D012639:Seeds / Q000378:metabolism; D013816:Thermodynamics
D000941:Antigens; D003619:Dansyl Compounds; D005456:Fluorescent Dyes; D037102:Lectins; D037121:Plant Lectins; C042242:Vicia lectins; D005688:Galactosamine; C041456:dansylgalactosamine