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 β-sheet, demonstrated to participate in interdomain interaction in phototropin. CD data show that YtvA-E105L has a lower α-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 β-sheet in the light-driven conformational changes in LOV proteins.
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1 September 2005
Mutational Effects on Protein Structural Changes and Interdomain Interactions in the Blue-light Sensing LOV Protein YtvA
Aba Losi,
Elisa Ghiraldelli,
Sven Jansen,
Wolfgang Gärtner
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Photochemistry and Photobiology
Vol. 81 • No. 5
September 2005
Vol. 81 • No. 5
September 2005