7QGK

The mRubyFT protein, Genetically Encoded Blue-to-Red Fluorescent Timer in its red state

7QGK の概要

• エントリーDOI: 10.2210/pdb7qgk/pdb
• 分子名称: The red form of the mRubyFT protein, Genetically Encoded Blue-to-Red Fluorescent Timer, MAGNESIUM ION (3 entities in total)
• 機能のキーワード: fluorescent protein, flourescent timer, mrubyft, blue to red timer, chromophore
• 由来する生物種: Entacmaea quadricolor
• タンパク質・核酸の鎖数: 1
• 化学式量合計: 27088.85

構造登録者

Boyko, K.M.,Nikolaeva, A.Y.,Gaivoronskii, F.A.,Vlaskina, A.V.,Subach, O.M.,Popov, V.O.,Subach, F.V. (登録日: 2021-12-08, 公開日: 2022-03-23, 最終更新日: 2024-02-07)

主引用文献

Subach, O.M.,Tashkeev, A.,Vlaskina, A.V.,Petrenko, D.E.,Gaivoronskii, F.A.,Nikolaeva, A.Y.,Ivashkina, O.I.,Anokhin, K.V.,Popov, V.O.,Boyko, K.M.,Subach, F.V.
The mRubyFT Protein, Genetically Encoded Blue-to-Red Fluorescent Timer.
Int J Mol Sci, 23:-, 2022

PubMed Abstract: Genetically encoded monomeric blue-to-red fluorescent timers (mFTs) change their fluorescent color over time. mCherry-derived mFTs were used for the tracking of the protein age, visualization of the protein trafficking, and labeling of engram cells. However, the brightness of the blue and red forms of mFTs are 2-3- and 5-7-fold dimmer compared to the brightness of the enhanced green fluorescent protein (EGFP). To address this limitation, we developed a blue-to-red fluorescent timer, named mRubyFT, derived from the bright mRuby2 red fluorescent protein. The blue form of mRubyFT reached its maximum at 5.7 h and completely transformed into the red form that had a maturation half-time of 15 h. Blue and red forms of purified mRubyFT were 4.1-fold brighter and 1.3-fold dimmer than the respective forms of the mCherry-derived Fast-FT timer in vitro. When expressed in mammalian cells, both forms of mRubyFT were 1.3-fold brighter than the respective forms of Fast-FT. The violet light-induced blue-to-red photoconversion was 4.2-fold less efficient in the case of mRubyFT timer compared to the same photoconversion of the Fast-FT timer. The timer behavior of mRubyFT was confirmed in mammalian cells. The monomeric properties of mRubyFT allowed the labeling and confocal imaging of cytoskeleton proteins in live mammalian cells. The X-ray structure of the red form of mRubyFT at 1.5 Å resolution was obtained and analyzed. The role of the residues from the chromophore surrounding was studied using site-directed mutagenesis.

PubMed: 35328628
DOI: 10.3390/ijms23063208

実験手法

X-RAY DIFFRACTION (1.5 Å)