Open AccessAn optomechanical transducer in the blue light receptor phototropin from Avena sativa
Author(s) -
Michael Salomon,
Wolfgang Eisenreich,
Harald Dürr,
Erik Schleicher,
Elke Knieb,
Vincent Massey,
Wolfhart Rüdiger,
Franz Müller,
Adelbert Bacher,
Gerald Richter
Publication year - 2001
Publication title -
proceedings of the national academy of sciences of the united states of america
Language(s) - English
Resource type - Journals
eISSN - 1091-6490
pISSN - 0027-8424
DOI - 10.1073/pnas.221455298
Subject(s) - phototropin , autophosphorylation , flavin group , histidine kinase , phototropism , pas domain , chemistry , biophysics , protein kinase domain , biochemistry , blue light , protein kinase a , photochemistry , biology , cryptochrome , kinase , histidine , materials science , enzyme , mutant , circadian clock , optoelectronics , transcription factor , gene
The PHOT1 (NPH1) gene from Avena sativa specifies the blue light receptor for phototropism, phototropin, which comprises two FMN-binding LOV domains and a serine/threonine protein kinase domain. Light exposure is conducive to autophosphorylation of the protein kinase domain. We have reconstituted a recombinant LOV2 domain of A. sativa phototropin with various (13)C/(15)N-labeled isotopomers of the cofactor, FMN. The reconstituted protein samples were analyzed by NMR spectroscopy under dark and light conditions. Blue light irradiation is shown to result in the addition of a thiol group (cysteine 450) to the 4a position of the FMN chromophore. The adduct reverts spontaneously in the dark by elimination. The light-driven flavin adduct formation results in conformational modification, which was diagnosed by (1)H and (31)P NMR spectroscopy. This conformational change is proposed to initiate the transmission of the light signal via conformational modulation of the protein kinase domain conducive to autophosphorylation of NPH1.