Photophysical characterisation and photo-cycle dynamics of LOV1-His domain of phototropin from Chlamydomonas reinhardtii with roseoflavin monophosphate cofactor

Journal of Photochemistry and Photobiology. B, Biology
A TyagiPeter Hegemann

Abstract

The wild-type phototropin protein phot from the green alga Chlamydomonas reinhardtii with the blue-light photoreceptor domains LOV1 and LOV2 has flavin mononucleotide (FMN) as cofactor. For the LOV1-His domain from phot of C. reinhardtii studied here, the FMN chromophore was replaced by roseoflavin monophosphate (8-dimethylamino-8-demethyl-FMN, RoFMN) during heterologous expression in a riboflavin auxotropic Escherichia coli strain. An absorption and emission spectroscopic characterisation of the cofactor exchanged-LOV1-His (RoLOV1) domain was carried out in aqueous pH 8 phosphate buffer. The fluorescence of RoLOV1 is quenched by photo-induced charge transfer at room temperature. The photo-cyclic dynamics of RoLOV1 was observed by blue-light induced hypochromic and bathochromic absorption changes which recover on a minute timescale in the dark. Photo-excited RoFMN is thought to cause reversible protein and cofactor structural changes. Prolonged intense blue-light exposure caused photo-degradation of RoFMN in RoLOV1 to fully reduced flavin and lumichrome derivatives. Photo-cycle schemes of RoLOV1 and LOV1 are presented, and the photo-degradation dynamics of RoLOV1 is discussed.

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Citations

Feb 11, 2014·Interface Focus·John T M Kennis, Tilo Mathes
Sep 13, 2014·The Journal of Physical Chemistry. B·Bora Karasulu, Walter Thiel
Nov 8, 2018·Photochemical & Photobiological Sciences : Official Journal of the European Photochemistry Association and the European Society for Photobiology·Yulia I MeteleshkoMaria G Khrenova
Mar 5, 2013·Chembiochem : a European Journal of Chemical Biology·Madina MansurovaWolfgang Gärtner
Jul 16, 2021·Journal of Photochemistry and Photobiology. B, Biology·Misato IkomaMasahide Terazima
Mar 11, 2011·The Journal of Physical Chemistry. a·N Shaemningwar Moyon, Sivaprasad Mitra

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