Equilibrium Unfolding of Bombyx mori Glycyl-tRNA Synthetase
作者： John David Dignam , Xiaogang Qu , Jonathan B. Chaires
关键词: Equilibrium unfolding 、 Native state 、 Urea 、 Quenching (fluorescence) 、 Chemistry 、 Crystallography 、 Mutant 、 Glycine—tRNA ligase 、 Tryptophan 、 Circular dichroism
摘要: Unfolding of Bombyx mori glycyl-tRNA synthetase was examined by multiple spectroscopic techniques. Tryptophan fluorescence wild type enzyme and an N-terminally truncated form (N55) increased at low concentrations urea or guanidine-HCl followed a reduction in intensity intermediate denaturant concentrations; transition higher detected as decreased red-shifted emission. Solute quenching indicated that tryptophans become progressively solvent-exposed during unfolding. Wild had stronger negative CD bands between 220 230 nm than the mutant, indicative greater alpha-helical content. Urea caused ellipticity 222 concentration with enzyme, is absent mutant; both enzymes exhibited cooperative concentrations. Both dissociate to monomers 1.5 m urea. described multistate unfolding parallel two state unfolding; two-state component mutant. Changes spectral properties associated were largely reversible after dilution denaturant. complex native state, native-like monomer, partially unfolded states, state.
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