An optomechanical transducer in the blue light receptor phototropin from <i>Avena sativa</i>

TLDR

The phototropin PHOT1 from *Avena sativa* contains two FMN‑binding LOV domains and a serine/threonine kinase domain that undergoes light‑induced autophosphorylation. The study proposes that the light‑induced conformational change in the LOV2 domain initiates signal transmission to the kinase domain, promoting autophosphorylation of NPH1. Recombinant LOV2 was reconstituted with 13C/15N‑labeled FMN and examined by NMR spectroscopy in dark and illuminated states. Blue light induces a cysteine‑450 thiol adduct at FMN’s 4a position that spontaneously reverts in the dark, and this adduct causes detectable conformational changes by 1H and 31P NMR.

Abstract

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 .