5DTZ

Crystal structure of rsFolder in the fluorescent on-state

5DTZ の概要

• エントリーDOI: 10.2210/pdb5dtz/pdb
• 分子名称: Green fluorescent protein, DI(HYDROXYETHYL)ETHER (3 entities in total)
• 機能のキーワード: fluorescent protein, gfp, reversibly switchable, cis chromophore
• 由来する生物種: Aequorea victoria (Jellyfish)
• タンパク質・核酸の鎖数: 4
• 化学式量合計: 117814.81

構造登録者

El Khatib, M.,Colletier, J.P.,Adam, V. (登録日: 2015-09-18, 公開日: 2016-01-27, 最終更新日: 2024-01-10)

主引用文献

El Khatib, M.,Martins, A.,Bourgeois, D.,Colletier, J.P.,Adam, V.
Rational design of ultrastable and reversibly photoswitchable fluorescent proteins for super-resolution imaging of the bacterial periplasm.
Sci Rep, 6:18459-18459, 2016

PubMed Abstract: Phototransformable fluorescent proteins are central to several nanoscopy approaches. As yet however, there is no available variant allowing super-resolution imaging in cell compartments that maintain oxidative conditions. Here, we report the rational design of two reversibly switchable fluorescent proteins able to fold and photoswitch in the bacterial periplasm, rsFolder and rsFolder2. rsFolder was designed by hybridisation of Superfolder-GFP with rsEGFP2, and inherited the fast folding properties of the former together with the rapid switching of the latter, but at the cost of a reduced switching contrast. Structural characterisation of the switching mechanisms of rsFolder and rsEGFP2 revealed different scenarios for chromophore cis-trans isomerisation and allowed designing rsFolder2, a variant of rsFolder that exhibits improved switching contrast and is amenable to RESOLFT nanoscopy. The rsFolders can be efficiently expressed in the E. coli periplasm, opening the door to the nanoscale investigation of proteins localised in hitherto non-observable cellular compartments.

PubMed: 26732634
DOI: 10.1038/srep18459

実験手法

X-RAY DIFFRACTION (1.5 Å)