Abstract
The binding of fluorescently labelled carbohydrates to concanavalin A and wheat germ agglutinin was studied at equilibrium and by the stopped-flow and temperature jump relaxation methods. Ligand were mainly die 4-methylumbelliferyl glycosides of α (1 → 2)-linked manno-oligosaccharides and of β (1 → 4)-linked chito oligosaccharides as limited homologous series. They offer distinct advantages, parti cularly for kinetic studies.
Enthalpie and kinetic considerations suggest that concanavalin A specifically binds a single mannopyranosyl group in α (1 →2)-linked manno-oligosaccharides. This occurs preferentially at the non-reducing end. Glycosylation of a carbohydrate withe.g. an aryl group does not afect die binding kinetics and for all carbohydrates the association rate is comparable but relatively slow, which indicates that a common process is involved in the binding of all carbohydrates to concanavalin A. The affinity of a carbohydrate for concanavalin A is determined by the dissociation-rate parameter, resulting in a longer residence time for a better ligand.
Interaction of chito-oligosaccharides with wheat germ agglutinin is complex. With the larger members of the 4-methylumbelliferyl chito-oligosaccharides, binding studies were only possible at low fractional saturation to avoid formation of unsoluble complexes. The binding kinetics of wheat germ agglutinin are faster than with concanavalin A and are consistent with a wheat germ agglutinin binding region composed of two adjacent subsites. For binding of the monoside as well as the bioside, two consistent kinetic models apply. They have common that for each ligand there exist two complexes with comparable population.
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Abbreviations
- MeUmb-Manp :
-
4-methylumbelliferyl α-D-mannopyranoside
- Con A:
-
concanavalin A
- WGA:
-
wheat germ agglutinin
- Me-Manp :
-
methyl-α-D-mannopyranoside
References
Allen, A. K., Neuberger, A. and Sharon, N. 1973Biochem J.,155, 127.
Bernasconi, C. F. (1976)Relaxation Kinetics, (New York: Academic Press) p. 22.
Bhavanandan, V. P. and Katlic, A. W. (1979)J. Biol. Chem.,254, 4000.
Brewer, C. F., Sternlicht, H., Marcus, D. M. and Grollman, A. P. (1973)Proc. Natl.Acad. Sci. USA,70, 1007.
Brewer, C. F., Marcus, D., Grollman, A. P. and Sternlicht, H. (1974) inLysozyme, eds. F. Osserman, et. al., (New York: Academic Press) p. 239.
Cardin, A. D:, Behnke, W. D. and Mandel, F. (1979)J. Biol. Chem.,254, 8877.
Christie, D. J., Alter, G. M. and Magnuson, J. A. (1978)Biochemistry,17, 4425.
Christie, D. J., Munske, G. R. and Magnuson, J. A. 1979Biochemistry,18, 4639.
Claeyssens, M. and De Bruyne, C. K. (1978)Biochim. Biophys. Acta,533, 98.
Clegg, R. M., Loontiens, F. G. and Jovin,T. M. (1977)Biochemistry,16, 167.
Clegg, R. M., Loontiens, F. G., Van Landschoot, A., Sharon, N., De Bruyne, C. K. and Jovin, T. M. 1980Arch. Int. Physiol. Biochem.,88, B69.
Clegg, R. M., Loontiens, F. G. Van Landschoot, A., Jovin, T. M. (1981)Biochemistry,20, 4687.
Dani, M., Manca, F. and Rialdi, G. (1981)Biochim. Biophys. Acta,667, 108.
Dean, B. R. and Horner, R. B. (1973)Biochim. Biophys. Acta,322 141.
De Boeck, H. (1980)Licentiaatsthesis, University of Ghent, Belgium.
De Boeck, H., Loontiens, F. G., Delmotte, F. M. and De Bruyne, C. K. (1981)FEBS Lett.,126, 227.
De Clercq, A., Loontiens, F. G. and De Bruyne, C. K. (1980)Biochem. Biophys.Res. Commun.,97, 364.
Delmotte, F. M., Privat, J. P. D. J. and Monsigny, M. L. P. (1975)Carbohydr. Res.,40, 353.
Erni, B., De Boeck, H., Loontiens, F. G. and Sharon, N. (1980)FEBS Lett.,120, 149.
Farina, R. D. and Wilkins, R. G. (1980)Biochim. Biophys. Acta,631, 428.
Fitos, I., Heremans, K. and Loontiens, F. G. (1979)Reaet. Kinet. Catal. Lett.,12, 393.
Goldstein, I. J., Hammarström, S. and Sundblad, G. (1975)Biochim. Biphys. Acta,405, 53.
Gray, R. D. and Glew, R. H. (1973)J. Biol. Chem.,248, 7547.
Grimaldi, J. J. and Sykes, B. D. (1975)Biol. Chem.,250, 1618.
Hansch, C. (1975)Adv. Pharmacol. Chemoter.,13, 45.
Hansch, C. and Calef, D. F. (1976)J. Org. Chem.,41, 1240.
Hardman, K. D. and Ainsworth, C. F. (1973)Biochemistry,12, 4442.
Harrington, P. C. and Wilkins, R. G. (1978)Biochemistry,17, 4245.
Harrington, P. C, Moreno, R. and Wilkins, R. G. (1981)Isr. J. Chem.,21, 48.
Khan, M. I., Mathew, M. K., Balaram, P. and Surolia, A. (1980)Biochem. J. 191, 395.
Khan, M. I., Mazumdar, T., Pain, D., Gaur, N. and Surolia, A. (1981a)Eur. J. Biochem.,113, 471.
Khan, M. I., Mathew, M. K., Mazumdar, T., Pain, D., Surolia, N., Das, M. K., Balaram, P. and Surolia, A. (1981b)Eur. J. Biochem.,115, 149.
Kieda, C. M. T., Delmotte, F. M. and Monsigny, M. L. P. (1977)Proc. Natl. Acad. Sci. USA,74,168.
Lewis, S. D. Shafer, J. A. and Goldstein, I. J. (1976)Arch. Biochem. Biophys.,172, 689.
Loontiens, F. G., Van Wauwe, J. P., De Gussem, R., De Bruyne, C. K. (1973)Carbohydr. Res.,30, 51.
Loontiens, F. G., Van Wauwe, J. P. and De Bruyne, C. K., (1975)Carbohydr. Res.,44, 150.
Loontiens, F. G., Clegg, R. M. and Jovin, T. M. (1977a)Biochemistry,16, 159.
Loontiens, F. G., Clegg, R. M., Van Landschoot, A. and, Jovin, T. M. (1977b)Eur. J. Biochem.,78, 465.
Lotan, R. and Sharon, N. (1973)Biochem. Biophys. Res. Commun.,55, 1340.
Midoux, P. (1980)Interactions: protéine-sucre;étude spectroscopique des interactions entre lα lectine du gerne de blé (WGA) etdes darivus de Iα N-acetyl-D-glucosamine, Orléans, Thèse 3ème cycle, Orléans.
Munske, G. R., Magnuson, J. A. and Krakauer, H. (1978)Biochem. Biophys. Res. Commun.,84, 684.
Nagata, Y. and, Burger, M. M. (1974)J. Biol. Chem.,249, 3116.
Pecht, I. and Lancet, D. (1977) inChemical Relaxations in Molecular Biology, eds I. Pecht and R. Rigler, (Berlin: Springer Verlag) p. 306.
Peters, B. P., Ebisu, S., Goldstein, I. J. and Flashner, M. (1979)Biochemistry,18, 5505.
Pflumm, M. R, Wang, J. L. and Edelman, G. M. (1971)J. Biol. Chem.,246, 4369.
Privat, J. P., Delmotte, F. and Monsigny, M. (1974a)FEBS Lett.,46, 224.
Privat, J. P., Delmotte, F. and Monsigny, M. (1974b)FEBS Lett.,46, 229.
Privat, J. P., Delmotte, F., Mialonier, G., Bouchard, P. and Monsigny, M. (1974c)Eur. J. Biochem.,47, 5.
Urdea, M. S., Christie, D. J., Munske, G. R., Magnuson, J. A. and Legg, J. I. (1979)Biochem. Biophys. Res. Commun.,95,1043.
Van Landschoot, A., Loontiens, F. G., Clegg, R. M., Sharon, N. and De Bruyne, C. K. (1977)Eur. J. Biochem.,79, 275.
Van Landschoot,A. (1978)Binding van Oligosacchariden en hun fluorescerende 4-methylumbelliferylderivaten op concanavaline A en op het lectine uit tarwekiemen;evenwichtsstudies, ‘stopped-flow’en temperatuursprongs-relaxatie kinetiek, Ghent Ph. D, thesis.
Van Landschoot, A., Loontiens, F. G. and De Bruyne, C. K. (1978a)Eur. J. Biochem.,83, 277.
Van Landschoot, A., Clegg, R. M., Loontiens, F. G. and Sharon, N. (1978b)9th Intl. Symp. Carbohydr. Chem. London, Abstract D18.
Van Landschoot, A., Loontiens, F. G., and De Bruyne, C. K. (1980a)Eur. J. Biochem.,103, 307.
Van Landschoot, A., Loontiens, F. G., Clegg, R. M. and Jovin, T. M. (1980b)Eur. J. Biochem.,103, 313.
Viale, R. D. (1971)J. Theor.Biol.,31, 501.
Williams, T. J., Shafer, J. A. and Goldstein, I. J. (1978a)J. Biol. Chem.,253, 8533.
Williams, T. J., Shafer, J. A., Goldstein, I. J. and Adamson, T. (1978b)J. Biol. Chem.,253, 8538.
Williams, T. J., Homer, L. D., Shafer, J. A., Goldstein, I. J., Garegg, P. J., Hultberg, H., Iversen, T. and Johansson, R. (1981)Arch. Biochem. Biophys.,209, 555.
Wright C. S. (1980a)J. Mol. Biol.,139, 53.
Wright, C. S. (1980b)J. Mol. Biol.,141, 267.
Yang, Y. and Hamaguchi, K. (1980)J. Biochem.,87, 1003.
Yang, Y. and Hamaguchi, K. (1980)J. Biochem.,88, 829.
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Loontiens, F.G., Clegg, R.M. & Van Landschoot, A. Some physicochemical aspects of oligosaccharide binding to concanavalin A and wheat germ agglutinin. J Biosci 5 (Suppl 1), 105–120 (1983). https://doi.org/10.1007/BF02702981
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DOI: https://doi.org/10.1007/BF02702981