Binding of 4-methyl umbelliferyl-α-D-glucoPyranoside to Vicia faba lectin: Fluorescence-quenching studies
On binding toVicia faba lectin, the fluorescence of 4-methylumbelliferyl-α-D-glucoPyranoside was quantitatively quenched showing that the interaction of 4-methylumbelliferyl-α-D-glucoPyranoside took Place in a binding environment. The binding of the fluorescent sugar was saccharide sPecific as evidenced by the reversal of 4-methylumbelliferyl-α-D-glucoPyranoside fluorescence quenching by D-fructose. The association constant,K a, values for the 4-methylumbelliferyl-α-D-glucoPyranoside was determined by comPetition study emPloying reversal of fluorescence quenching of 4-methylumbelliferyl-α-D-glucoPyranoside by D-fructose. TheK a value obtained for D-fructose was 1.07 ±0.03 X 104 M-1 and for 4-methylumbelliferyl-α-D-glucoPyranoside was 1.60 ±0.05 X 104 M-1 at 15°C. TheK a values of 2.51 ±0.06 X 104M-1, l.26 ±0.02 X 104 M-1 and 0.56 ±0.01 X 104M-1, resPectively at 10°, 20° and 30°C were obtained from the ChiPman equation. The relative fluorescence quenching, ΔF a, at infinite concentration of the free saccharide sites ofVicia faba lectin [P′] was 93.5% at 30°C and the binding constant for 4-methylumbelliferyl-α-D-glucoPyranoside lectin interaction as derived by Yank and Hanaguchi equation was 0.63 ±0.01 X 104M-1.
KeywordsVicia faba lectin lectin-saccharide binding fluorescence quenching binding constants
Bessler, W., Shafer, J. A. and Goldstein, I. J. (1974)J. Biol. Chem.
, 2819.PubMedGoogle Scholar
de Boeck, H., Loontiens, F. G., Delmotte, F. M. and de Bruyne, C. K. (1981)FEBS Lett.
, 227.PubMedCrossRefGoogle Scholar
Chipman, D. M., Grisaro, U. and Sharon, N. (1967)J. Biol. Chem.
, 4388.PubMedGoogle Scholar
Datta, P. K., Basu, P. S. and Datta, T. K. (1984a)PreP. Biochem.
, 373.PubMedCrossRefGoogle Scholar
Datta, P. K., Basu, P. S. and Datta, T. K. (1984b)IRCSMed. Sci.
, 687.Google Scholar
Datta, P. K., Basu, P. S. and Datta, T. K. (1984c)Indian J. Biochem. BioPhys.
, 174.PubMedGoogle Scholar
Datta, P. K., Basu, P. S. and Datta, T. K. (1986)Carbohydr. Res.
, 259.CrossRefGoogle Scholar
Dean, B. R. and Homer, R. B. (1973)Biochem. BioPhys. Acta.
, 141.PubMedGoogle Scholar
Debray, H., Decout, D., Strecher, G., SPik, G. and Montreuil, J. (1981)Eur. J. Biochem.
, 41.PubMedCrossRefGoogle Scholar
Goldstein, I. J. and Hayes, C. E. (1978)Adv. Carbohydr. Chem. Biochem.
, 128.CrossRefGoogle Scholar
Katagiri, Y., Yamamoto, K., Tsuji, T., Osawa, T. (1984)Carbohydr. Res.
, 257.CrossRefGoogle Scholar
Loontiens, F-G., Clegg, R. M. and Jovin, T. M. (1977)Biochemistry
, 159.PubMedCrossRefGoogle Scholar
Mertens, M. L. and Kagi, J. H. R. (1979)Anal. Biochem.
, 448.PubMedCrossRefGoogle Scholar
Monique, D., Michel, V., Khushi, M. L. and Jean-Pierre, F. (1984)Arch. Biochem. BioPhys.
, 640.CrossRefGoogle Scholar
Privat, J. P., Delmotte, F. and Monsigny, M. (1974)FEBS Lett.
, 229.PubMedCrossRefGoogle Scholar
SiPs, R. (1948)J. Chem. Phys.
, 490.CrossRefGoogle Scholar
ThomPson, R. B. and Lakowiez, J. R. (1984)Biochim. BioPhys. Acta
, 87.PubMedGoogle Scholar
van Landschoot, A., Loontiens, F. G., Clegg, R. M., Sharon, N. and DeBruyne, C. K. (1977)Eur. J. Biochem.
, 275.PubMedCrossRefGoogle Scholar
Yank, Y. and Hamaguchi, K. (1980)J. Biochem.
, 829.Google Scholar
© Indian Academy of Sciences 1987