Abstract
The binding of the apolar fluorescent dye 8-anilinonaphthalene-1-sulfonate (ANS) to bovine serum albumin (BSA), phospholipase A2 (PLA2), ovalbumin, lysozyme, cobrotoxin and N-acetyltryptophanamide was used to assess the factors affecting the efficiency of energy transfer from Trp residues to the ANS molecule. We found that the efficiency of energy transfer from Trp residues to ANS was associated with the ability of proteins to enhance the ANS fluorescence. At the same molar concentration of protein, BSA enhanced ANS fluorescence most among these proteins; its Trp fluorescence was drastically quenched by the addition of ANS. Fluorescence enhancement of ANS in PLA2-ANS complex increased upon addition of Ca2+ or change of the buffer to acidicpH, resulting in a higher efficiency of energy transfer from Trp residues to ANS. There was limited ANS fluorescence enhancement with ovalbumin, lysozyme, cobrotoxin, and N-acetyltryptophanamide and a less efficient quenching in Trp fluorescence. The capabilities of proteins for binding with ANS correlated with the decrease in their Trp fluorescence being quenching by ANS. However, the microenvironment surrounding Trp residues of proteins did not affect the energy transfer. Based on these results, the factors that affected the energy transfer from Trp residues to ANS are discussed.
Similar content being viewed by others
References
Bashford, C. L., and Harris, D. A. (1987). InSpectrophotometry & Spectrofluorimetry, IRL Press, Oxford, pp. 91–114.
Bradford, M. (1976).Anal. Biochem. 72, 248–254.
Campbell, I. D., and Dwek, R. A. (1984). InBiological Spectroscopy, Benjamin/Cummings, Menlo Park, California, pp. 91–126.
Cardamone, M., and Puri, N. K. (1992).Biochem. J. 282, 589–593.
Chang, L. S., and Yang, C. C. (1993).J. Protein Chem. 12, 579–583.
Chang, L. S., Kuo, K. W., and Chang, C. C. (1993).Biochim. Biophys. Acta 1202, 216–220.
Johnson, J. D., El-Bayoumi, A. E., Weber, L. D., and Tulinsky, A. (1979).Biochemistry 18, 1292–1296.
Kollmann-Koch, A., and Eggerer, H. (1989).Eur. J. Biochem. 185, 441–447.
Kuo, K. W., and Chang, C. C. (1991).J. Biochem. (Tokyo)110, 863–867.
Lacowicz, J. R. (1983). InPrinciples of Fluorescence Spectroscopy, Plenum Press, New York, pp. 305–341.
Purdon, A. D., Tinker, D. O., and Spero, L. (1977).Can. J. Biochem. 55, 205–214.
Stryer, L. (1968).Science 162, 526–533.
Waite, M. (1987). InPhospholipase, Plenum Press, New York, pp. 155–241.
Yang, C. C., and Chang, L. S. (1984).J. Protein Chem. 3, 195–213.
Author information
Authors and Affiliations
Rights and permissions
About this article
Cite this article
Chang, Ls., Wen, Ey., Hung, Jj. et al. Energy transfer from tryptophan residues of proteins to 8-anilinonaphthalene-1-sulfonate. J Protein Chem 13, 635–640 (1994). https://doi.org/10.1007/BF01890462
Received:
Published:
Issue Date:
DOI: https://doi.org/10.1007/BF01890462