Abstract
In continuation of our preceding study, this report describes pH and ionic strength dependences of the binding constants of six anionic azo dyes to lysozyme, the competitive binding of the dyes with the substrate analogues of lysozyme and the change in the circular dichroism of lysozyme by the dye binding. The binding constants were obtained from the difference spectra of visible absorption. With an increase in pH from 5.0 to 7.0 the constants for the dyes (1st of the two modes of the binding of a dye named D3) are reduced. The increase in ionic strength from 0.1 to 0.2 also reduces the values of the constant. Competitive binding was found between D3 and the analogues, but not for the other dyes. The change in the circular dichroism due to the electronic perturbation of tryptophyl residues in lysozyme was found. From these evidences, Lys 33 in lysozyme is pinpointed as the most credible binding site for the dyes (1st mode binding of D3). An unspecified location near the subsiteB in lysozyme is addressed for 2nd mode binding of D3.
Similar content being viewed by others
References
Nakano, Y., T. Seki, H. Iwata, J. Komiyama, T. Iijima, Colloid and Polymer Sci., in press?
Rupley, J. A., Biochim. Biophys. Acta.83, 245 (1964).
Sophianopoulous, A. J., C. K. Rhodes, D. N. Holcomb, K. E. Van Holde, J. Biol. Chem.237, 1107 (1962).
Holler, E., J. A. Rupley, G. P. Hess, Biochemistry14, 1088 (1975).
Benesi, H. A., J. H. Hildebrand, J. Amer. Chem. Soc.71, 2703 (1949).
Sakakibara, R., K. Hamaguchi, J. Biochem. (Tokyo)64, 613 (1968).
Tanford, C., M. L. Wagner, J. Amer. Chem. Soc.76, 3331 (1954).
Imoto, T., L. N. Johnson, A. C. T. North, D. C. Phillips, J. A. Rupley, “The Enzymes”, vol. VII (P. D. Boyer, ed.), pp. 665, Academic Press, New York (1972).
Perkins, S. J., L. N. Johnson, D. C. Phillips, R. A. Dwek, Biochem. J.193 553 (1981).
Horwitz, J., E. H. Strickland, C. Billups, J. Amer. Chem. Soc.91, 184 (1969).
Ikeda, K., K. Hamaguchi, J. Biochem. (Tokyo)71, 265 (1972).
Author information
Authors and Affiliations
Rights and permissions
About this article
Cite this article
Nakano, Y., Kawauchi, S., Komiyama, J. et al. The interaction of anionic azo dyes with hen egg-white lysozyme: 2. Colloid & Polymer Sci 260, 339–344 (1982). https://doi.org/10.1007/BF01447974
Received:
Published:
Issue Date:
DOI: https://doi.org/10.1007/BF01447974