title: A conserved isoleucine in the LOV1 domain of a novel phototropin from the marine alga Ostreococcus tauri modulates the dark state recovery of the domain.
abstract: BACKGROUND
  Phototropins are UV-A/blue light receptor proteins with two LOV (Light-Oxygen-Voltage) sensor domains at their N terminus and a kinase domain at the C-terminus in photoautotrophic organisms. This is the first research report of a canonical phototropin from marine algae Ostreococcus tauri.
  
  
  METHODS
  We synthesized core LOV1 (OtLOV1) domain-encoding portion of the phototropin gene of O. tauri, the domain was heterologously expressed, purified and assessed for its spectral properties and dark recovery kinetics by UV-Visible, fluorescence spectroscopy and mutational studies. Quaternary structure characteristics were studied by SEC and glutaraldehyde crosslinking.
  
  
  RESULTS
  The absorption spectrum of OtLOV1 lacks the characteristic 361nm peak shown by other LOV1 domains. It undergoes a photocycle with a dark state recovery time of approximately 30min (τ=300.35s). Native OtLOV1 stayed as dimer in aqueous solution and the dimer formation was light and concentration independent. Mutating isoleucine at 43rd position to valine accelerated the dark recovery time by more than 10-fold. Mutating it to serine reduced sensitivity to blue light, but the dark recovery time remained unaltered. I43S mutation also destabilized the FMN binding to a great extent.
  
  
  CONCLUSIONS
  The OtLOV1 domain of the newly identified OtPhot is functional and the isoleucine at position 43 of OtLOV1 is the key residue responsible for fine-tuning the domain properties.
  
  
  CONCLUSIONS
  This is the first characterized LOV1 domain of a canonical phototropin from a marine alga and spectral properties of the domain are similar to that of the LOV1 domain of higher plants.
authors.affiliation: Department of Biochemistry, University of Delhi South Campus, Benito Juares Road, New Delhi-110021, India.
authors.forename: Sindhu Kandoth
authors.initials: SK
authors.lastname: Veetil
authors.forename: Chitvan
authors.initials: C
authors.lastname: Mittal
authors.forename: Peeyush
authors.initials: P
authors.lastname: Ranjan
authors.forename: Suneel
authors.initials: S
authors.lastname: Kateriya
mesh_terms: D000595:Amino Acid Sequence; D001665:Binding Sites / Q000235:genetics; D000460:Chlorophyta / Q000235:genetics / Q000378:metabolism*; D003001:Cloning, Molecular; D003624:Darkness; D004591:Electrophoresis, Polyacrylamide Gel; D005486:Flavin Mononucleotide / Q000378:metabolism; D008958:Models, Molecular; D008969:Molecular Sequence Data; D009154:Mutation; D056949:Phototropins / Q000737:chemistry* / Q000235:genetics / Q000378:metabolism*; D055503:Protein Multimerization; D017434:Protein Structure, Tertiary; D011994:Recombinant Proteins / Q000737:chemistry / Q000378:metabolism; D012623:Seawater / Q000382:microbiology; D017422:Sequence Analysis, DNA; D017386:Sequence Homology, Amino Acid; D013050:Spectrometry, Fluorescence; D013053:Spectrophotometry
chemical_list: D056949:Phototropins; D011994:Recombinant Proteins; D005486:Flavin Mononucleotide