title: 13C Isotopologue editing of FMN bound to phototropin domains.
abstract: The plant blue light receptor phototropin comprises a protein kinase domain and two FMN-binding LOV domains (LOV1 and LOV2). Blue light irradiation of recombinant LOV domains is conducive to the addition of a cysteinyl thiolate group to carbon 4a of the FMN chromophore, and spontaneous cleavage of that photoadduct completes the photocycle of the receptor. The present study is based on (13)C NMR signal modulation observed after reconstitution of LOV domains of different origins with random libraries of (13)C-labeled FMN isotopologues. Using this approach, all (13)C signals of FMN bound to LOV1 and LOV2 domains of Avena sativa and to the LOV2 domain of the fern, Adiantum capillus-veneris, could be unequivocally assigned under dark and under blue light irradiation conditions. (13)C Chemical shifts of FMN are shown to be differently modulated by complexation with the LOV domains under study, indicating slight differences in the binding interactions of FMN and the apoproteins.
authors.affiliation: Lehrstuhl für Organische Chemie und Biochemie, Technische Universität München, Garching, Germany. wolfgang.eisenreich@ch.tum.de
authors.forename: Wolfgang
authors.initials: W
authors.lastname: Eisenreich
authors.forename: Monika
authors.initials: M
authors.lastname: Joshi
authors.forename: Boris
authors.initials: B
authors.lastname: Illarionov
authors.forename: Gerald
authors.initials: G
authors.lastname: Richter
authors.forename: Werner
authors.initials: W
authors.lastname: Römisch-Margl
authors.forename: Franz
authors.initials: F
authors.lastname: Müller
authors.forename: Adelbert
authors.initials: A
authors.lastname: Bacher
authors.forename: Markus
authors.initials: M
authors.lastname: Fischer
mesh_terms: D000595:Amino Acid Sequence; D001483:Base Sequence; D002247:Carbon Isotopes / Q000737:chemistry; D056931:Cryptochromes; D004247:DNA; D005486:Flavin Mononucleotide / Q000737:chemistry / Q000378:metabolism*; D005420:Flavoproteins / Q000737:chemistry / Q000378:metabolism*; D009682:Magnetic Resonance Spectroscopy; D008958:Models, Molecular; D008969:Molecular Sequence Data; D017386:Sequence Homology, Amino Acid; D012689:Sequence Homology, Nucleic Acid
chemical_list: D002247:Carbon Isotopes; D056931:Cryptochromes; D005420:Flavoproteins; D005486:Flavin Mononucleotide; D004247:DNA