Light-Driven Domain Mechanics of a Minimal Phytochrome Photosensory Module Studied by EPR

Abstract

Light-exposed organisms developed photoreceptors to transduce light signals for environmental adaptation. Phytochromes, found in bacteria, fungi, and plants, can discriminate the ratio of red and far-red light using the isomerization of a bilin chromophore bound to a photosensory module to trigger downstream conformational changes in the protein. Here, we investigated by hydrogen/deuterium exchange mass spectrometry and electron paramagnetic resonance spectroscopy the light-driven domain mechanics of a minimal monomeric photosensory module from the group II phytochrome Cph2 from Synechocystis sp. PCC 6803. We could unambiguously trace the light-driven secondary structural rearrangement of its tongue region, and we found a translational motion of the PHY domain that is related to what was found before by X-ray studies in a group I module. Our analysis demonstrates a common light response in the photosensory modules of phytochromes, orchestrated solely by the GAF-PHY bidomain independent of further quaternary interactions or the nature of downstream effector domains.

Keywords: DEER; EPR spectroscopy; HDX mass spectrometry; bilin chromophore; cyanobacterial phytochrome; domain mechanics; elastic network modeling; secondary structure; β/α transition.