title: Microsecond light-induced proton transfer to flavin in the blue light sensor plant cryptochrome.
abstract: Plant cryptochromes are blue light photoreceptors that regulate key responses in growth and daily rhythm of plants and might be involved in magnetoreception. They show structural homology to the DNA repair enzyme photolyase and bind flavin adenine dinucleotide as chromophore. Blue light absorption initiates the photoreduction from the oxidized dark state of flavin to the flavin neutral radical, which is the signaling state of the sensor. Previous time-resolved studies of the photoreduction process have been limited to observation of the decay of the radical in the millisecond time domain. We monitored faster, light-induced changes in absorption of an algal cryptochrome covering a spectral range of 375-750 nm with a streak camera setup. Electron transfer from tryptophan to flavin is completed before 100 ns under formation of the flavin anion radical. Proton transfer takes place with a time constant of 1.7 micros leading to the flavin neutral radical. Finally, the flavin radical and a tryptophan neutral radical decay with a time constant >200 micros in the millisecond and second time domain. The microsecond proton transfer has not been observed in animal cryptochromes from insects or photolyases. Furthermore, the strict separation in time of electron and proton transfer is novel in the field of flavin-containing photoreceptors. The reaction rate implies that the proton donor is not in hydrogen bonding distance to the flavin N5. Potential candidates for the proton donor and the involvement of the tryptophan triad are discussed.
authors.affiliation: Institut für Physikalische und Theoretische Chemie, Universität Regensburg, 93053 Regensburg, Germany.
authors.forename: Thomas
authors.initials: T
authors.lastname: Langenbacher
authors.forename: Dominik
authors.initials: D
authors.lastname: Immeln
authors.forename: Bernhard
authors.initials: B
authors.lastname: Dick
authors.forename: Tilman
authors.initials: T
authors.lastname: Kottke
mesh_terms: D000818:Animals; D017360:Arabidopsis / Q000737:chemistry / Q000378:metabolism; D029681:Arabidopsis Proteins; D016825:Chlamydomonas reinhardtii / Q000737:chemistry / Q000378:metabolism; D056931:Cryptochromes; D005182:Flavin-Adenine Dinucleotide / Q000737:chemistry* / Q000378:metabolism; D005420:Flavoproteins / Q000737:chemistry* / Q000378:metabolism; D008958:Models, Molecular; D010084:Oxidation-Reduction; D017434:Protein Structure, Tertiary; D011522:Protons; D014364:Tryptophan / Q000737:chemistry / Q000378:metabolism
chemical_list: D029681:Arabidopsis Proteins; C538962:CRY1 protein, Arabidopsis; D056931:Cryptochromes; D005420:Flavoproteins; D011522:Protons; D005182:Flavin-Adenine Dinucleotide; D014364:Tryptophan