8BGL

Structure of the dimeric rsCherryRev1.4

8BGL の概要

• エントリーDOI: 10.2210/pdb8bgl/pdb
• 関連するPDBエントリー: 8B65 8B7G
• 分子名称: PAmCherry1 protein, DI(HYDROXYETHYL)ETHER, PENTAETHYLENE GLYCOL, ... (5 entities in total)
• 機能のキーワード: dimerization, reversibly switchable, red fluorescent proteins, disulfide bond, fluorescent protein
• 由来する生物種: Discosoma sp.
• タンパク質・核酸の鎖数: 2
• 化学式量合計: 61595.45

構造登録者

Bui, T.Y.H.,Van Meervelt, L. (登録日: 2022-10-28, 公開日: 2023-02-15, 最終更新日: 2024-10-16)

主引用文献

Bui, T.Y.H.,Dedecker, P.,Van Meervelt, L.
An unusual disulfide-linked dimerization in the fluorescent protein rsCherryRev1.4.
Acta Crystallogr.,Sect.F, 79:38-44, 2023

PubMed Abstract: rsCherryRev1.4 has been reported as one of the reversibly photoswitchable variants of mCherry, and is an improved version with a faster off-switching speed and lower switching fatigue at high light intensities than its precursor rsCherryRev. However, rsCherryRev1.4 still has some limitations such as a tendency to dimerize as well as complex photophysical properties. Here, the crystal structure of rsCherryRev1.4 was determined at a resolution of 2 Å and it was discovered that it forms a dimer that shows disulfide bonding between the protomers. Mutagenesis, gel electrophoresis and size-exclusion chromatography strongly implicate Cys24 in this process. Replacing Cys24 in rsCherryRev1.4 resulted in a much lower tendency towards dimerization, while introducing Cys24 into mCherry correspondingly increased its dimerization. In principle, this finding opens the possibility of developing redox sensors based on controlled dimerization via disulfide cross-linking in fluorescent proteins, even though the actual application of engineering such sensors still requires additional research.

PubMed: 36748340
DOI: 10.1107/S2053230X23000572

実験手法

X-RAY DIFFRACTION (2 Å)