EMDB/PDB/SASBDBから引用されている文献のデータベース

キーワード
構造解析手法	All X線回折 NMR 電子顕微鏡小角散乱中性子線回折線維回折粉末回折赤外分光 EPR FRET 理論モデルハイブリッド
著者・登録者
ジャーナル
IF	>30 >20 >10 >5 all

タイトル	Near-atomic cryo-EM imaging of a small protein displayed on a designed scaffolding system.
ジャーナル・号・ページ	Proc Natl Acad Sci U S A, Vol. 115, Issue 13, Page 3362-3367, Year 2018
掲載日	2018年3月27日
著者	Yuxi Liu / Shane Gonen / Tamir Gonen / Todd O Yeates /
PubMed 要旨	Current single-particle cryo-electron microscopy (cryo-EM) techniques can produce images of large protein assemblies and macromolecular complexes at atomic level detail without the need for crystal ...Current single-particle cryo-electron microscopy (cryo-EM) techniques can produce images of large protein assemblies and macromolecular complexes at atomic level detail without the need for crystal growth. However, proteins of smaller size, typical of those found throughout the cell, are not presently amenable to detailed structural elucidation by cryo-EM. Here we use protein design to create a modular, symmetrical scaffolding system to make protein molecules of typical size suitable for cryo-EM. Using a rigid continuous alpha helical linker, we connect a small 17-kDa protein (DARPin) to a protein subunit that was designed to self-assemble into a cage with cubic symmetry. We show that the resulting construct is amenable to structural analysis by single-particle cryo-EM, allowing us to identify and solve the structure of the attached small protein at near-atomic detail, ranging from 3.5- to 5-Å resolution. The result demonstrates that proteins considerably smaller than the theoretical limit of 50 kDa for cryo-EM can be visualized clearly when arrayed in a rigid fashion on a symmetric designed protein scaffold. Furthermore, because the amino acid sequence of a DARPin can be chosen to confer tight binding to various other protein or nucleic acid molecules, the system provides a future route for imaging diverse macromolecules, potentially broadening the application of cryo-EM to proteins of typical size in the cell.
リンク	Proc Natl Acad Sci U S A / PubMed:29507202 / PubMed Central
手法	EM (単粒子)
解像度	3.09 - 4.03 Å
構造データ	EMDB-7403: Single-particle reconstruction of DARP14 - A designed protein scaffold displaying ~17kDa DARPin proteins - Helical extension 手法: EM (単粒子) / 解像度: 4.03 Å 7436 6c9i EMDB-7436, PDB-6c9i: Single-Particle reconstruction of DARP14 - A designed protein scaffold displaying ~17kDa DARPin proteins - Scaffold 手法: EM (単粒子) / 解像度: 3.09 Å 7437 6c9k EMDB-7437, PDB-6c9k: Single-Particle reconstruction of DARP14 - A designed protein scaffold displaying ~17kDa DARPin proteins 手法: EM (単粒子) / 解像度: 3.49 Å
由来	synthetic construct (人工物) Pyrococcus horikoshii OT3 (古細菌) Pseudomonas aeruginosa PAO1 (緑膿菌) pyrococcus horikoshii (strain atcc 700860 / dsm 12428 / jcm 9974 / nbrc 100139 / ot-3) (古細菌) pseudomonas aeruginosa (strain atcc 15692 / dsm 22644 / cip 104116 / jcm 14847 / lmg 12228 / 1c / prs 101 / pao1) (緑膿菌) synthetic (人工物)
キーワード	DE NOVO PROTEIN / Designed Complex / DARPin / Scaffold / Single-Particle Cryo-EM