TAILIEUCHUNG - Báo cáo khoa học: C Isotopologue editing of FMN bound to phototropin domains

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. | ễFEBS Journal 13C Isotopologue editing of FMN bound to phototropin domains Wolfgang Eisenreich1 Monika Joshi1 Boris Illarionov1 Gerald Richter2 Werner Romisch-Margl1 Franz Muller3 Adelbert Bacher1 and Markus Fischer4 1 Lehrstuhl fur Organische Chemie und Biochemie Technische Universitat Munchen Garching Germany 2 Schoolof Chemistry Cardiff University UK 3 Wylstrasse 13 Hergiswil Switzerland 4 Institut fur Biochemie und Lebensmittelchemie Abteilung Lebensmittelchemie Universitat Hamburg Germany Keywords blue light receptor isotopologue libraries LOV domain NMR spectroscopy phototropin Correspondence W. Eisenreich Lehrstuhl fur Organische Chemie und Biochemie Technische Universitat Munchen Lichtenbergstrasse 4 D-85747 Garching Germany Fax 49 89 289 13363 Tel 49 89 289 13336 E-mail M. Fischer Institut fur Biochemie und Lebensmittelchemie Abteilung Lebensmittelchemie Universitat Hamburg Grindelallee 117 20146 Hamburg Germany Fax 49 40 4283 84342 Tel 49 40 4283 84357 E-mail Received 6 June 2007 revised 12 August 2007 accepted 19 September 2007 doi 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 13C NMR signal modulation observed after reconstitution of LOV domains of different origins with random libraries of 13C-labeled FMN isotopologues. Using this approach all 13C 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. 13C Chemical shifts of FMN are shown to be differently modulated by

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