5NFX

Deinococcus radiodurans BphP PAS-GAF Y263F mutant

5NFX の概要

• エントリーDOI: 10.2210/pdb5nfx/pdb
• 分子名称: Bacteriophytochrome, ACETATE ION, (4S)-2-METHYL-2,4-PENTANEDIOL, ... (5 entities in total)
• 機能のキーワード: kinase, photosensor, transferase, phytochrome
• 由来する生物種: Deinococcus radiodurans R1
• タンパク質・核酸の鎖数: 1
• 化学式量合計: 38094.43

構造登録者

Takala, H.,Westenhoff, S.,Ihalainen, J.A. (登録日: 2017-03-16, 公開日: 2018-04-18, 最終更新日: 2024-11-13)

主引用文献

Takala, H.,Lehtivuori, H.K.,Berntsson, O.,Hughes, A.,Nanekar, R.,Niebling, S.,Panman, M.,Henry, L.,Menzel, A.,Westenhoff, S.,Ihalainen, J.A.
On the (un)coupling of the chromophore, tongue interactions, and overall conformation in a bacterial phytochrome.
J. Biol. Chem., 293:8161-8172, 2018

PubMed Abstract: Phytochromes are photoreceptors in plants, fungi, and various microorganisms and cycle between metastable red light-absorbing (Pr) and far-red light-absorbing (Pfr) states. Their light responses are thought to follow a conserved structural mechanism that is triggered by isomerization of the chromophore. Downstream structural changes involve refolding of the so-called tongue extension of the phytochrome-specific GAF-related (PHY) domain of the photoreceptor. The tongue is connected to the chromophore by conserved DIP and PRSF motifs and a conserved tyrosine, but the role of these residues in signal transduction is not clear. Here, we examine the tongue interactions and their interplay with the chromophore by substituting the conserved tyrosine (Tyr) in the phytochrome from the extremophile bacterium with phenylalanine. Using optical and FTIR spectroscopy, X-ray solution scattering, and crystallography of chromophore-binding domain (CBD) and CBD-PHY fragments, we show that the absence of the Tyr hydroxyl destabilizes the β-sheet conformation of the tongue. This allowed the phytochrome to adopt an α-helical tongue conformation regardless of the chromophore state, hence distorting the activity state of the protein. Our crystal structures further revealed that water interactions are missing in the Y263F mutant, correlating with a decrease of the photoconversion yield and underpinning the functional role of Tyr in phytochrome conformational changes. We propose a model in which isomerization of the chromophore, refolding of the tongue, and globular conformational changes are represented as weakly coupled equilibria. The results also suggest that the phytochromes have several redundant signaling routes.

PubMed: 29622676
DOI: 10.1074/jbc.RA118.001794

実験手法

X-RAY DIFFRACTION (1.34 Å)