Yk. Reshetnyak et Ea. Burstein, ASSIGNMENT OF LOG-NORMAL COMPONENTS OF FLUORESCENCE-SPECTRA OF SERINEPROTEASES TO THE CLUSTERS OF TRYPTOPHAN RESIDUES, Biofizika, 42(4), 1997, pp. 785-795
The two log-normal spectral components in, alpha-chymotrypsin, trypsin
ogen and beta-trypsin fluorescence and the single component of chymotr
ypsinogen A emission were analyzed using characteristics of microenvir
onment of atoms of indolic fluorophores of individual tryptophan resid
ues in the three-dimensional structures of the proteins. Tryptophan re
sidues (eight ones in chymotrypsinogen A and alpha-chymotrypsin and fo
ur ones in trypsinogen and beta-trypsin) in these homologous proteins
form three clusters with resonance excitation energy exchange between
fluorophores within each of them. In all the proteins, Trp51 and 141 d
etermine the shorter-wavelength spectral components (ca. 330 nm) and T
rp215 and 237 emit as longer-wavelength ones (ca. 340 nm). Excited sta
tes of fluorophores of Trp27, 29, 207 (cluster A) and 172 (in cluster
B) are deactivated by the near disulfide bonds. The longer-wavelength
component is quenched in chymotrypsinogen emission due probably to the
formation of an electron trap including Asn91 amide and several water
molecules. The contribution of longer-wavelength component in trypsin
spectrum increases comparing with that of trypsinogen due to some lit
tle changes in localization and a huge rise of mobility of Met180 sulf
ur atom nearly Trp215 in trypsin.