Startseite The amino acids surrounding the flavin 7a-methyl group determine the UVA spectral features of a LOV protein
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The amino acids surrounding the flavin 7a-methyl group determine the UVA spectral features of a LOV protein

  • Sarah Raffelberg , Alexander Gutt , Wolfgang Gärtner , Carmen Mandalari , Stefania Abbruzzetti , Cristiano Viappiani und Aba Losi EMAIL logo
Veröffentlicht/Copyright: 3. Juli 2013

Abstract

Flavin-binding light, oxygen, and voltage (LOV) domains are UVA/blue-light-sensing protein units that form a reversible flavin mononucleotide-cysteine adduct upon light induction. In their dark-adapted state, LOV domains exhibit the typical spectral features of fully oxidized riboflavin derivatives. A survey on the absorption spectra of various LOV domains revealed that the UVA spectral range is the most variable region (whereas the absorption band at 450 nm is virtually unchanged), showing essentially two distinct patterns found in plant phototropin LOV1 and LOV2 domains, respectively. In this work, we have identified a residue directly interacting with the isoalloxazine methyl group at C(7a) as the major UVA spectral tuner. In YtvA from Bacillus subtilis, this amino acid is threonine 30, and its mutation into apolar residues converts the LOV2-like spectrum of native YtvA into a LOV1-like pattern. Mutation T30A also accelerates the photocycle ca. 4-fold. Together with control mutations at different positions, our results experimentally confirm the previously calculated direction of the transition dipole moment for the UVA ππ* state and identify the mechanisms underlying spectral tuning in the LOV domains.


Corresponding author: Aba Losi, Department of Physics and Earth Sciences, University of Parma (www.unipr.it), Viale G.P. Usberti 7/A, I-43124, Parma, Italy, e-mail:

S.R. is a recipient of a PhD student grant from the ‘Biostruct’ program of the Heinrich-Heine-University Düsseldorf. This work has been partially supported by the Vigoni program (to A.L. and W.G.) and the University of Parma (fellowship to C.M.). We thank Francesca Pennacchietti for the solvatochromic data on FMN reported within the supplementary material.

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Received: 2013-4-23
Accepted: 2013-7-1
Published Online: 2013-07-03
Published in Print: 2013-11-01

©2013 by Walter de Gruyter Berlin Boston

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