S. Vangala et al., Probing the relation between protein structure and intrinsic tryptophan fluorescence using superrepressor mutants of the trp repressor, J FLUORESC, 8(1), 1998, pp. 1-11
The wavelength dependence of the intrinsic tryptophan fluorescence lifetime
of a series of mutants of the trp repressor protein was characterized in b
oth the native and the denatured states. These mutants belong to a particul
ar class, called superrepressors, as their phenotype, when expressed in viv
o, is to repress transcription at lower concentrations of the corepressor,
tryptophan. II has been demonstrated previously that these mutations result
in distinct and profound modifications of the structural and dynamic: prop
erties of the protein [Reedstrom and Royer (1995) J. Mel. Biol. 253, 266; R
eedstrom et al. (1996) J. Mel. Biol. 263, 32; Smith et nl. (1995) Biochemis
try 34, 13183]. The present observations reveal that in the native state, t
hese structural and dynamic modifications result in subtle, yet significant
alterations in the intrinsic tryptophan fluorescence decay characteristics
. Surprisingly, significant differences in the fluorescence decays between
the mutants and the wild-type protein were also observed for the guanidine
hydrochloride unfolded states. These results are discussed in terms of the
various models which have been proposed to explain the decay properties of
tryptophan in proteins.