Nanosecond-pulse fluorimetry of wheat germ agglutinin is analyzed as a function of both excitation and emission wavelengths. When excited at 280 nm, wheat germ agglutinin fluorescence exhibited three lifetimes: one corresponding to the tyrosine residues as a whole and two others corresponding to the tryptophyl emission. The tyrosine contribution to the emission spectrum deduced from this method was in good agreement with that reported previously in steady-state fluorescence experiments [Privat, J.P. and Monsigny, M. (1975) Eur. J. Biochem. 60, 555-567]. The fluorescence decay of each tryptophan residue was not a single exponential function when wheat germ agglutinin was excited at 295 nm. This could be related to the microenvironment of the indole chromophores in the protein. The comparison of the quantum yield and of average lifetime showed that some tryptophan residues were completely quenched. Energy transfer from tyrosines to tryptophan residues previously detected in steady-state fluorescence was also revealed by fluorescence decay measurements. Comparison of both methods showed that an important part of transfers occurred with a very fast rate equal to or greater than 10(10) s-1. Both lifetimes and the ratio of the short and the long-lived component were found dependent on tri-N-acetylchitotriose binding.