4XVP

X-ray structure of bGFP-C / EGFP complex

4XVP の概要

• エントリーDOI: 10.2210/pdb4xvp/pdb
• 関連するPDBエントリー: 4XL5
• 分子名称: Green fluorescent protein, BGFP-C (2 entities in total)
• 機能のキーワード: alpharep scaffold, complex, egfp, protein engineering, heat-like repeat, protein binding, fluorescent protein
• 由来する生物種: Aequorea victoria (Jellyfish); 詳細
• タンパク質・核酸の鎖数: 6
• 化学式量合計: 148490.23

構造登録者

Chevrel, A.,Urvoas, A.,Li de la Sierra-Gallay, I.,Van Tilbeurgh, H.,Minard, P.,Valerio-Lepiniec, M. (登録日: 2015-01-27, 公開日: 2015-08-19, 最終更新日: 2026-03-18)

主引用文献

Chevrel, A.,Urvoas, A.,de la Sierra-Gallay, I.L.,Aumont-Nicaise, M.,Moutel, S.,Desmadril, M.,Perez, F.,Gautreau, A.,van Tilbeurgh, H.,Minard, P.,Valerio-Lepiniec, M.
Specific GFP-binding artificial proteins ( alpha Rep): a new tool for in vitro to live cell applications.
Biosci.Rep., 35:-, 2015

PubMed Abstract: A family of artificial proteins, named αRep, based on a natural family of helical repeat was previously designed. αRep members are efficiently expressed, folded and extremely stable proteins. A large αRep library was constructed creating proteins with a randomized interaction surface. In the present study, we show that the αRep library is an efficient source of tailor-made specific proteins with direct applications in biochemistry and cell biology. From this library, we selected by phage display αRep binders with nanomolar dissociation constants against the GFP. The structures of two independent αRep binders in complex with the GFP target were solved by X-ray crystallography revealing two totally different binding modes. The affinity of the selected αReps for GFP proved sufficient for practically useful applications such as pull-down experiments. αReps are disulfide free proteins and are efficiently and functionally expressed in eukaryotic cells: GFP-specific αReps are clearly sequestrated by their cognate target protein addressed to various cell compartments. These results suggest that αRep proteins with tailor-made specificity can be selected and used in living cells to track, modulate or interfere with intracellular processes.

PubMed: 26182430
DOI: 10.1042/BSR20150080

実験手法

X-RAY DIFFRACTION (3.4 Å)