title: Mutational effects on protein structural changes and interdomain interactions in the blue-light sensing LOV protein YtvA.
abstract: Mutagenesis studies on the phototropin-related protein YtvA from Bacillus subtilis have revealed the role of selected structural elements in interdomain communication. The LOV (light, oxygen, voltage) domain of YtvA undergoes light-driven reactions similar to that of phot-LOV, with reversible formation of a covalent flavin-cysteine adduct. The mutated proteins Ytva-E105L and YtvA-E56Q have been studied by UV fluorescence and circular dichroism (CD) spectroscopy. E105 (L in phototropin) is located at the solvent-exposed surface of the LOV domain central beta-sheet, demonstrated to participate in interdomain interaction in phototropin. CD data show that YtvA-E105L has a lower alpha-helix content in the dark and undergoes larger light-driven conformational changes than YtvA-WT. The E56Q mutation breaks the E56-K97 salt bridge, a structural element highly conserved within the LOV series. In YtvA-E56Q the CD spectrum is the same as in YtvA-WT, although the conserved W103 becomes more exposed to the solvent and the dark-recovery kinetics is slower. These results indicate that the E56-K97 salt bridge stabilizes locally the protein structure and participates in the regulation of the photocycle but has negligible effects on the overall structure. The E105L mutation, instead, highlights the involvement of the central beta-sheet in the light-driven conformational changes in LOV proteins.
authors: Department of Physics, University of Parma, Italy.
authors: Aba
authors: A
authors: Losi
authors: Elisa
authors: E
authors: Ghiraldelli
authors: Sven
authors: S
authors: Jansen
authors: Wolfgang
authors: W
authors: Gärtner
mesh_terms: D001412:Bacillus subtilis / Q000528:radiation effects*; D001426:Bacterial Proteins / Q000235:genetics* / Q000528:radiation effects*; D001483:Base Sequence; D008027:Light; D008958:Models, Molecular; D008969:Molecular Sequence Data; D009154:Mutation; D017434:Protein Structure, Tertiary / Q000235:genetics / Q000528:radiation effects; D013057:Spectrum Analysis
chemical_list: D001426:Bacterial Proteins