Item

Abstract

A prokaryotic protein, YtvA from Bacillus subtilis, was found to possess a light, oxygen, voltage (LOV) domain sharing high homology with the photoactive, flavin mononucleotide (FMN)- binding LOV domains of phototropins (phot), blue-light photoreceptors for phototropism in higher plants. Computer- based three-dimensional modeling suggests that YtvA-LOV binds FMN in a similar pocket as phot-LOVs. Recombinant YtvA indeed exhibits the same spectroscopical features and blue-light- induced photochemistry as phot-LOVs, with the reversible formation of a blue-shifted photoproduct, assigned to an FMN- cysteine thiol adduct (Thio(383)). By means of laser-flash photolysis and time-resolved optoacoustic experiments, we measured the quantum yield of formation for Thio(383), Phi(Thio) = 0.49, and the enthalpy change, DeltaH(Thio) = 135 kJ/mol, with respect to the parent state. The formation of Thio(383) is accompanied by a considerable volume contraction, DeltaV(Thio) = -13.5 ml/mol. Similar to phot-LOVs, Thio(383) is formed from the decay of a red-shifted transient species, T- 650, within 2 mus. In both YtvA and free FMN, this transient has an enthalpy content of similar to200 kJ/mol, and its formation is accompanied by a small contraction, DeltaV(T) approximate to -1.5 ml/mol, supporting the assignment of T-650 to the FMN triplet state, as suggested by spectroscopical evidences. These are the first studies indicating that phototropin-related, blue-light receptors may exist also in prokaryotes, besides constituting a steadily growing family in plants.