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	Coupled catalytic states and the role of metal coordination in Cas9.
ジャーナル・号・ページ	Nat Catal, Vol. 6, Issue 10, Page 969-977, Year 2023
掲載日	2023年10月2日
著者	Anuska Das / Jay Rai / Mitchell O Roth / Yuerong Shu / Megan L Medina / Mackenzie R Barakat / Hong Li /
PubMed 要旨	Controlling the activity of the CRISPR-Cas9 system is essential to its safe adoption for clinical and research applications. Although the conformational dynamics of Cas9 are known to control its ...Controlling the activity of the CRISPR-Cas9 system is essential to its safe adoption for clinical and research applications. Although the conformational dynamics of Cas9 are known to control its enzymatic activity, details of how Cas9 influences the catalytic processes at both nuclease domains remain elusive. Here we report five cryo-electron microscopy structures of the active Cas9 complex along the reaction path at 2.2-2.9 Å resolution. We observed that a large movement in one nuclease domain, triggered by the cognate DNA, results in noticeable changes in the active site of the other domain that is required for metal coordination and catalysis. Furthermore, the conformations synchronize the reaction intermediates, enabling coupled cutting of the two DNA strands. Consistent with the roles of conformations in organizing the active sites, adjustments to the metal-coordination residues lead to altered metal specificity of Cas9 and commonly used Cas9 in cells.
リンク	Nat Catal / PubMed:38348449 / PubMed Central
手法	EM (単粒子)
解像度	2.21 - 2.88 Å
構造データ	27141 8d2k EMDB-27141, PDB-8d2k: Structure of Acidothermus cellulolyticus Cas9 ternary complex (Cleavage Intermediate 2) 手法: EM (単粒子) / 解像度: 2.43 Å 27142 8d2l EMDB-27142, PDB-8d2l: Structure of Acidothermus cellulolyticus Cas9 ternary complex (Cleavage Intermediate 1) 手法: EM (単粒子) / 解像度: 2.21 Å 27143 8d2n EMDB-27143, PDB-8d2n: Structure of Acidothermus cellulolyticus Cas9 ternary complex (Pre-cleavage) 手法: EM (単粒子) / 解像度: 2.88 Å 27144 8d2o EMDB-27144, PDB-8d2o: Structure of Acidothermus cellulolyticus Cas9 ternary complex (Post-cleavage 2) 手法: EM (単粒子) / 解像度: 2.66 Å 27145 8d2p EMDB-27145, PDB-8d2p: Structure of Acidothermus cellulolyticus Cas9 ternary complex (Target bound) 手法: EM (単粒子) / 解像度: 2.78 Å 27146 8d2q EMDB-27146, PDB-8d2q: Structure of Acidothermus cellulolyticus Cas9 ternary complex (Post-cleavage 1) 手法: EM (単粒子) / 解像度: 2.58 Å
化合物	ChemComp-MG: Unknown entry / マグネシウムジカチオン ChemComp-HOH: WATER
由来	acidothermus cellulolyticus 11b (バクテリア) acidothermus cellulolyticus (バクテリア) synthetic construct (人工物)
キーワード	RNA BINDING PROTEIN/DNA/RNA / Cas9 / AcCas9 / Crispr / RNA BINDING PROTEIN / RNA BINDING PROTEIN-DNA-RNA complex / Post-cleavage 2 / AceCas9 / Target bound / Post-Cleavage 1