3U8A

Crystal structure of monomeric reversibly photoswitchable red fluorescent protein rsTagRFP in the OFF state

3U8A の概要

• エントリーDOI: 10.2210/pdb3u8a/pdb
• 関連するPDBエントリー: 3M22 3U8C
• 分子名称: Fluorescent protein rsTagRFP (2 entities in total)
• 機能のキーワード: beta-barrel, fluorescent protein, light induced reversible photoswitching, reversibly photoswitchable fluorescent protein, cis-trans isomerization
• 由来する生物種: synthetic construct (artificial gene)
• タンパク質・核酸の鎖数: 4
• 化学式量合計: 110557.82

構造登録者

Pletnev, S. (登録日: 2011-10-16, 公開日: 2012-02-22, 最終更新日: 2025-03-26)

主引用文献

Pletnev, S.,Subach, F.V.,Dauter, Z.,Wlodawer, A.,Verkhusha, V.V.
A structural basis for reversible photoswitching of absorbance spectra in red fluorescent protein rsTagRFP.
J.Mol.Biol., 417:144-151, 2012

PubMed Abstract: rsTagRFP is the first monomeric red fluorescent protein (FP) with reversibly photoswitchable absorbance spectra. The switching is realized by irradiation of rsTagRFP with blue (440 nm) and yellow (567 nm) light, turning the protein fluorescence ON and OFF, respectively. It is perhaps the most useful probe in this color class that has yet been reported. Because of the photoswitchable absorbance, rsTagRFP can be used as an acceptor in photochromic Förster resonance energy transfer. Yellow FPs, YPet and mVenus, are demonstrated to be excellent photochromic Förster resonance energy transfer donors for the rsTagRFP acceptor in its fusion constructs. Analysis of X-ray structures has shown that photoswitching of rsTagRFP is accompanied by cis-trans isomerization and protonation/deprotonation of the chromophore, with the deprotonated cis- and protonated trans-isomers corresponding to its ON and OFF states, respectively. Unlike in other photoswitchable FPs, both conformers of rsTagRFP chromophore are essentially coplanar. Two other peculiarities of the rsTagRFP chromophore are an essentially hydrophobic environment of its p-hydroxyphenyl site and the absence of direct hydrogen bonding between this moiety and the protein scaffold. The influence of the immediate environment on rsTagRFP chromophore was probed by site-directed mutagenesis. Residues Glu145 and His197 were found to participate in protonation/deprotonation of the chromophore accompanying the photoswitching of rsTagRFP fluorescence, whereas residues Met160 and Leu174 were shown to spatially restrict chromophore isomerization, favoring its radiative decay.

PubMed: 22310052
DOI: 10.1016/j.jmb.2012.01.044

実験手法

X-RAY DIFFRACTION (1.783 Å)