6MWQ

Single particle cryoEM structure of a DARPin-aldolase platform in complex with GFP

6MWQ の概要

• エントリーDOI: 10.2210/pdb6mwq/pdb
• EMDBエントリー: 9277
• 分子名称: DARPin, Muscle-type aldolase chimeric fusion, Green fluorescent protein (2 entities in total)
• 機能のキーワード: synthetic construct, platform, single particle cryoem, small protein display, lyase-fluorescent protein complex, lyase/fluorescent protein
• 由来する生物種: synthetic construct; 詳細
• タンパク質・核酸の鎖数: 8
• 化学式量合計: 314348.52

構造登録者

Weaver, S.J.,Yao, Q. (登録日: 2018-10-30, 公開日: 2018-11-21, 最終更新日: 2024-03-13)

主引用文献

Yao, Q.,Weaver, S.J.,Mock, J.Y.,Jensen, G.J.
Fusion of DARPin to Aldolase Enables Visualization of Small Protein by Cryo-EM.
Structure, 27:1148-, 2019

PubMed Abstract: Solving protein structures by single-particle cryoelectron microscopy (cryo-EM) has become a crucial tool in structural biology. While exciting progress is being made toward the visualization of small macromolecules, the median protein size in both eukaryotes and bacteria is still beyond the reach of cryo-EM. To overcome this problem, we implemented a platform strategy in which a small protein target was rigidly attached to a large, symmetric base via a selectable adapter. Of our seven designs, the best construct used a designed ankyrin repeat protein (DARPin) rigidly fused to tetrameric rabbit muscle aldolase through a helical linker. The DARPin retained its ability to bind its target: GFP. We solved the structure of this complex to 3.0 Å resolution overall, with 5-8 Å resolution in the GFP region. As flexibility in the DARPin position limited the overall resolution of the target, we describe strategies to rigidify this element.

PubMed: 31080120
DOI: 10.1016/j.str.2019.04.003

実験手法

ELECTRON MICROSCOPY (3 Å)