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	Polyelectrolyte coating of cryo-EM grids improves lateral distribution and prevents aggregation of macromolecules.
ジャーナル・号・ページ	Acta Crystallogr D Struct Biol, Vol. 78, Issue Pt 11, Page 1337-1346, Year 2022
掲載日	2022年11月1日
著者	Dominik Hrebík / Mária Gondová / Lucie Valentová / Tibor Füzik / Antonín Přidal / Jiří Nováček / Pavel Plevka /
PubMed 要旨	Cryo-electron microscopy (cryo-EM) is one of the primary methods used to determine the structures of macromolecules and their complexes. With the increased availability of cryo-electron microscopes, ...Cryo-electron microscopy (cryo-EM) is one of the primary methods used to determine the structures of macromolecules and their complexes. With the increased availability of cryo-electron microscopes, the preparation of high-quality samples has become a bottleneck in the cryo-EM structure-determination pipeline. Macromolecules can be damaged during the purification or preparation of vitrified samples for cryo-EM, making them prone to binding to the grid support, to aggregation or to the adoption of preferential orientations at the air-water interface. Here, it is shown that coating cryo-EM grids with a negatively charged polyelectrolyte, such as single-stranded DNA, before applying the sample reduces the aggregation of macromolecules and improves their distribution. The single-stranded DNA-coated grids enabled the determination of high-resolution structures from samples that aggregated on conventional grids. The polyelectrolyte coating reduces the diffusion of macromolecules and thus may limit the negative effects of the contact of macromolecules with the grid support and blotting paper, as well as of the shear forces on macromolecules during grid blotting. Coating grids with polyelectrolytes can readily be employed in any laboratory dealing with cryo-EM sample preparation, since it is fast, simple, inexpensive and does not require specialized equipment.
リンク	Acta Crystallogr D Struct Biol / PubMed:36322417 / PubMed Central
手法	EM (単粒子)
解像度	1.77 - 3.8 Å
構造データ	14679 7ze1 EMDB-14679, PDB-7ze1: Tribolium castaneum hexamerin 2 手法: EM (単粒子) / 解像度: 3.2 Å EMDB-14680: Tribolium castaneum hexamerin 2 from aggregated sample 手法: EM (単粒子) / 解像度: 3.1 Å 14704 7zfw EMDB-14704, PDB-7zfw: Structure of human 80S ribosome obtained from ssDNA coated grid 手法: EM (単粒子) / 解像度: 3.8 Å 14705 7zg7 EMDB-14705, PDB-7zg7: Structure of human Apoferritin obtained from ssDNA coated grid 手法: EM (単粒子) / 解像度: 1.77 Å
化合物	ChemComp-ZN: Unknown entry ChemComp-NA: Unknown entry / ナトリウムカチオン ChemComp-HOH: WATER / 水
由来	tribolium castaneum (コクヌストモドキ) red flour beetle (コクヌストモドキ) homo sapiens (ヒト)
キーワード	UNKNOWN FUNCTION / hexamerin / tribolium castaneum / storage protein (貯蔵タンパク質) / RIBOSOME (リボソーム) / human 80S ribosome / ssDNA covered grid / ssDNA (デオキシリボ核酸) / cryoEM (低温電子顕微鏡法) / METAL BINDING PROTEIN / apoferritin (フェリチン)