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	Cryo-Electron Microscopy and Mass Analysis of Oligolysine-Coated DNA Nanostructures.
ジャーナル・号・ページ	ACS Nano, Vol. 15, Issue 6, Page 9391-9403, Year 2021
掲載日	2021年6月22日
著者	Eva Bertosin / Pierre Stömmer / Elija Feigl / Maximilian Wenig / Maximilian N Honemann / Hendrik Dietz /
PubMed 要旨	Cationic coatings can enhance the stability of synthetic DNA objects in low ionic strength environments such as physiological fluids. Here, we used single-particle cryo-electron microscopy (cryo-EM), ...Cationic coatings can enhance the stability of synthetic DNA objects in low ionic strength environments such as physiological fluids. Here, we used single-particle cryo-electron microscopy (cryo-EM), pseudoatomic model fitting, and single-molecule mass photometry to study oligolysine and polyethylene glycol (PEG)-oligolysine-coated multilayer DNA origami objects. The coatings preserve coarse structural features well on a resolution of multiple nanometers but can also induce deformations such as twisting and bending. Higher-density coatings also led to internal structural deformations in the DNA origami test objects, in which a designed honeycomb-type helical lattice was deformed into a more square-lattice-like pattern. Under physiological ionic strength, where the uncoated objects disassembled, the coated objects remained intact but they shrunk in the helical direction and expanded in the direction perpendicular to the helical axis. Helical details like major/minor grooves and crossover locations were not discernible in cryo-EM maps that we determined of DNA origami coated with oligolysine and PEG-oligolysine, whereas these features were visible in cryo-EM maps determined from the uncoated reference objects. Blunt-ended double-helical interfaces remained accessible underneath the coating and may be used for the formation of multimeric DNA origami assemblies that rely on stacking interactions between blunt-ended helices. The ionic strength requirements for forming multimers from coated DNA origami differed from those needed for uncoated objects. Using single-molecule mass photometry, we found that the mass of coated DNA origami objects prior to and after incubation in low ionic strength physiological conditions remained unchanged. This finding indicated that the coating effectively prevented strand dissociation but also that the coating itself remained stable in place. Our results validate oligolysine coatings as a powerful stabilization method for DNA origami but also reveal several potential points of failure that experimenters should watch to avoid working with false premises.
リンク	ACS Nano / PubMed:33724780 / PubMed Central
手法	EM (単粒子)
解像度	10.38 - 21.28 Å
構造データ	EMDB-12467: B-brick 0.2:1 N:P PEG-oligolysine coated in 5 mM Mg2+ 手法: EM (単粒子) / 解像度: 12.0 Å EMDB-12468: B-brick 100:1 N:P PEG-oligolysine coated in 5 mM Mg2+ 手法: EM (単粒子) / 解像度: 21.28 Å EMDB-12469: B-brick 100:1 N:P PEG-oligolysine coated in PBS 手法: EM (単粒子) / 解像度: 16.95 Å EMDB-12470: B-brick 0.75:1 N:P PEG-oligolysine coated in 5 mM Mg2+ 手法: EM (単粒子) / 解像度: 14.95 Å EMDB-12471: B-brick 0.75:1 N:P PEG-oligolysine coated in PBS 手法: EM (単粒子) / 解像度: 18.76 Å EMDB-12472: B-brick 0.5:1 N:P oligolysine coated in 5 mM Mg2+ 手法: EM (単粒子) / 解像度: 16.26 Å EMDB-12473: A-brick bare in 5 mM Mg2+ 手法: EM (単粒子) / 解像度: 11.88 Å EMDB-12474: A-brick 0.75:1 N:P PEG-oligolysine coated in 5 mM Mg2+ 手法: EM (単粒子) / 解像度: 12.42 Å EMDB-12475: A-brick 0.75:1 N:P PEG-oligolysine coated in PBS 手法: EM (単粒子) / 解像度: 16.29 Å EMDB-12476: A-brick 0.5:1 N:P oligolysine coated in 5 mM Mg2+ 手法: EM (単粒子) / 解像度: 17.7 Å 12516 7npn EMDB-12516, PDB-7npn: B-brick bare in 5 mM Mg2+ 手法: EM (単粒子) / 解像度: 10.38 Å
由来	synthetic construct (人工物)
キーワード	DNA / DNA Origami