Abstract
The binding of benzo(α)pyrene, ellipticine, and cis-parinaric acid to native, esterified, and alkylated β-lactoglobulin was followed by enhancement of the ligand fluorescence. Three studied ligands bind to native or modified β-lactoglobulin in apparent molar ratios varying between 1/8 and 2/1, with apparent dissociation constants in the range of 10−8 M for ligand/β-lactoglobulin complexes. The studied, chemically modified β-lactoglobulin derivatives display higher binding affinities for all studied ligands, cis-parinaric acid excluded. The reductive alkylation of ε-NH2 lysyl residues of β-lactoglobulin increases the apparent molar ratios of benzo(α)pyrene and cis-parinaric acid, and decreases it for ellipticine. The esterified and native β-lactoglobulin complexed to the investigated ligands display similar stoichiometries. Dynamic light scattering study of ligand-β-lactoglobulin complexes in solution shows the formation of aggregates: the apparent hydrodynamic radius value of β-lactoglobulin dimer (3.4 nm) reaches 49, 46, and 74 nm upon addition and binding of benzo(α)pyrene, ellipticine, and cis-parinaric acid, respectively.
Similar content being viewed by others
References
Adler-Nissen, J. (1979).J. Agric. Food Chem. 27, 1256–1262.
Berde, C., Hudson, B., Simoni, R., and Sklar, L. (1979).J. Biol. Chem. 254, 391–400.
Bertrand-Harb, C., Chobert, J. M., Dufour, E., and Haertlé, T. (1991).Sci. Aliments 11, 641–652.
Brown, E. D. (1984).J. Dairy Sci. 67, 713–722.
Bull, H. B., and Breese, K. (1967).Arch. Biochem. Biophys. 120, 309–315.
Cabacungan, J. C., Ahmed, A. I., and Feeney, R. E. (1982).Anal. Biochem. 124, 272–278.
Cogan, U., Kopelman, M., Mokady, S., and Shinitzky, M. (1976).Eur. J. Biochem. 65, 71–78.
Diaz de Villegas, M. C., Oria, R., Sala, F. G., and Calvo, M. (1987).Milchwissenschaft 42, 357–358.
Dodin, G., Schwaller, M. A., Aubard, J., and Paoletti, C. (1988).Eur. J. Biochem. 176, 371–376.
Dodin, G., Andrieux, M., and Al Kabbani, H. (1990).Eur. J. Biochem. 193, 697–700.
Dufour, E., and Haertlé, T. (1990).J. Agric. Food Chem. 38, 1691–1695.
Dufour, E., and Haertlé, T. (1991).Biochim. Biophys. Acta 1079, 316–320.
Dufour, E., Marden, M. C., and Haertlé, T. (1990).FEBS Lett. 277, 223–226.
Farrell, H. M., Behe, M. J., and Enyart, J. A. (1987).J. Dairy Sci. 70, 252–258.
Fraenkel-Conrat, H., and Olcott, H. S. (1945).J. Biol. Chem. 161, 259–268.
Fugate, R. D., and Song, P. S. (1980).Biochim. Biophys. Acta 625, 28–42.
Futterman, S., and Heller, J. (1972).J. Biol. Chem. 247, 5168–5172.
Godovac-Zimmerman, J. (1988).Trends in Biochem. Sci. 13, 64–66.
Grinberg, N., Blanco, R., Yarmush, D. M., and Karger, B. L. (1989).Anal. Chem. 61, 514–520.
Huber, R., Schneider, M., Epp, O., Mayr, I., Messerschmidt, A., Pflugrath, J., and Kayser, H. (1987).J. Mol. Biol. 195, 423–434.
Koppel, D. E. (1972).J. Chem. Phys. 57, 4814–4817.
Le Gal, J. M., and Manfait, M. (1990).Biochim. Biophys. Acta 1041, 257–263.
Mailliart, P., and Ribadeau-Dumas, B. (1988).J. Food Sci. 53, 743–745.
Monaco, H. L., Zanotti, G., Spadon, P., Bolognesi, M., Sawyer, L., and Eliopoulos, E. E. (1987).J. Mol. Biol. 197, 695–706.
Monti, J. C., Mermoud, A. F., and Jolles, P. (1989).Experientia 45, 178–180.
Moulti-Mati, F., Mati, A., Capiaumont, J., Belleville, F., Linden, G., and Nabet, P. (1991).Le Lait 71, 543–553.
Newcomer, M. E., Jones, T. A., Aqvist, J., Sundelin, J., Eriksson, U., Rask, L., and Peterson, P. (1984).EMBO J. 3, 1451–1454.
North, A. C. T. (1989).Int. J. Biol. Macromol. 11, 56–58.
Ono, J., Doi, K., Ogasa, K., and Nagasawa, T. (1975).Agr. Biol. Chem. 39, 2149–2155.
Papiz, M. Z., Sawyer, L., Eliopoulos, E. E., North, A. C. T., Findlay, J. B. C., Sivaprasadarao, R., Jones, T. A., Newcomer, M. E., and Kraulis P. J. (1986).Nature 324, 383–385.
Pessen, H., Purcell, J. M., and Farrell, H. M. (1985).Biochim. Biophys. Acta 828, 1–12.
Said, H. M., Ong, D. E., and Shingleton, J. L. (1989).Am. J. Clin. Nutr. 49, 690–694.
Sawyer, L., and Richardson, J. S. (1991).Trends in Biochem. Sci. 16, 11.
Timasheff, S. N., and Townend, R. (1964).Nature 203, 517–519.
Westphal, U. (1969).Methods Enzymol. 15, 761–796.
Zimmerman, K. D., Barlow, G. H., and Klotz, I. M. (1970).Arch. Biochem. Biophys. 138, 101–109.
Author information
Authors and Affiliations
Rights and permissions
About this article
Cite this article
Dufour, E., Roger, P. & Haertlé, T. Binding of benzo(α)pyrene, ellipticine, and cis-parinaric acid to β-lactoglobulin: Influence of protein modifications. J Protein Chem 11, 645–652 (1992). https://doi.org/10.1007/BF01024965
Received:
Published:
Issue Date:
DOI: https://doi.org/10.1007/BF01024965