title: FMN binding and photochemical properties of plant putative photoreceptors containing two LOV domains, LOV/LOV proteins.
abstract: LOV domains function as blue light-sensing modules in various photoreceptors in plants, fungi, algae, and bacteria. A LOV/LOV protein (LLP) has been found from Arabidopsis thaliana (AtLLP) as a two LOV domain-containing protein. However, its function remains unknown. We isolated cDNA clones coding for an LLP homolog from tomato (Solanum lycopersicum) and two homologs from the moss Physcomitrella patens. The tomato LLP (SlLLP) contains two LOV domains (LOV1 and LOV2 domains), as in AtLLP. Most of the amino acids required for association with chromophore are conserved in both LOV domains, except that the amino acid at the position equivalent to the cysteine essential for cysteinyl adduct formation is glycine in the LOV1 domain as in AtLLP. When expressed in Escherichia coli, SlLLP binds FMN and undergoes a self-contained photocycle upon irradiation of blue light. Analyses using mutant SlLLPs revealed that SlLLP binds FMN in both LOV domains, although the LOV1 domain does not show spectral changes on irradiation. However, when Gly(66) in the LOV1 domain, which is located at the position equivalent to the essential cysteine of LOV domains, is replaced by cysteine, the mutated LOV1 domain shows light-induced spectral changes. In addition, all four LOV domains of P. patens LLPs (PpLLP1 and PpLLP2) show the typical features of LOV domains, including the reactive cysteine in each. This study shows that plants have a new LOV domain-containing protein family with the typical biochemical and photochemical properties of other LOV domain-containing proteins such as the phototropins.
authors.affiliation: Department of Biotechnology, College of Life Sciences, Ritsumeikan University, Nojihigashi, Kusatsu, Shiga 525-8577, Japan. kasa@sk.ritsumei.ac.jp
authors.forename: Masahiro
authors.initials: M
authors.lastname: Kasahara
authors.forename: Mayumi
authors.initials: M
authors.lastname: Torii
authors.forename: Akimitsu
authors.initials: A
authors.lastname: Fujita
authors.forename: Kengo
authors.initials: K
authors.lastname: Tainaka
mesh_terms: D000595:Amino Acid Sequence; D017360:Arabidopsis / Q000235:genetics / Q000378:metabolism; D019068:Bryopsida / Q000235:genetics / Q000378:metabolism*; D005486:Flavin Mononucleotide / Q000235:genetics / Q000378:metabolism*; D008027:Light; D018551:Solanum lycopersicum; D008969:Molecular Sequence Data; D056949:Phototropins / Q000235:genetics / Q000378:metabolism*; D011485:Protein Binding; D017434:Protein Structure, Tertiary; D011994:Recombinant Proteins / Q000235:genetics / Q000378:metabolism
chemical_list: D056949:Phototropins; D011994:Recombinant Proteins; D005486:Flavin Mononucleotide
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