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	Cryoelectron microscopy of Na,K-ATPase in the two E2P states with and without cardiotonic steroids.
ジャーナル・号・ページ	Proc Natl Acad Sci U S A, Vol. 119, Issue 15, Page e2123226119, Year 2022
掲載日	2022年4月12日
著者	Ryuta Kanai / Flemming Cornelius / Bente Vilsen / Chikashi Toyoshima /
PubMed 要旨	Cryoelectron microscopy (cryo-EM) was applied to Na+,K+-ATPase (NKA) to determine the structures of two E2P states, one (E2PATP) formed by ATP and Mg2+ in the forward reaction, and the other (E2PPi) ...Cryoelectron microscopy (cryo-EM) was applied to Na+,K+-ATPase (NKA) to determine the structures of two E2P states, one (E2PATP) formed by ATP and Mg2+ in the forward reaction, and the other (E2PPi) formed by inorganic phosphate (Pi) and Mg2+ in the backward reaction, with and without ouabain or istaroxime, representatives of classical and new-generation cardiotonic steroids (CTSs). These two E2P states exhibit different biochemical properties. In particular, K+-sensitive acceleration of the dephosphorylation reaction is not observed with E2PPi, attributed to the presence of a Mg2+ ion in the transmembrane cation binding sites. The cryo-EM structures of NKA demonstrate that the two E2P structures are nearly identical but Mg2+ in the transmembrane binding cavity is identified only in E2PPi, corroborating the idea that it should be denoted as E2PPi·Mg2+. We can now explain why the absence of transmembrane Mg2+ in E2PATP confers the K+ sensitivity in dephosphorylation. In addition, we show that ATP bridges the actuator (A) and nucleotide binding (N) domains, stabilizing the E2PATP state; CTS binding causes hardly any changes in the structure of NKA, both in E2PATP and E2PPi·Mg2+, indicating that the binding mechanism is conformational selection; and istaroxime binds to NKA, extending its aminoalkyloxime group deep into the cation binding site. This orientation is upside down compared to that of classical CTSs with respect to the steroid ring. Notably, mobile parts of NKA are resolved substantially better in the electron microscopy (EM) maps than in previous X-ray structures, including sugars sticking out from the β-subunit and many phospholipid molecules.
リンク	Proc Natl Acad Sci U S A / PubMed:35380894 / PubMed Central
手法	EM (単粒子) / X線回折
解像度	2.9 - 3.9 Å
構造データ	32894 7wyu EMDB-32894, PDB-7wyu: Cryo-EM structure of Na+,K+-ATPase in the E2P state formed by ATP 手法: EM (単粒子) / 解像度: 3.4 Å 32895 7wyv EMDB-32895, PDB-7wyv: Cryo-EM structure of Na+,K+-ATPase in the E2P state formed by ATP in the presence of 40 mM Mg2+ 手法: EM (単粒子) / 解像度: 3.7 Å 32896 7wyw EMDB-32896, PDB-7wyw: Cryo-EM structure of Na+,K+-ATPase in the E2P state formed by inorganic phosphate 手法: EM (単粒子) / 解像度: 3.8 Å 32897 7wyx EMDB-32897, PDB-7wyx: Cryo-EM structure of Na+,K+-ATPase in the E2P state formed by ATP with istaroxime 手法: EM (単粒子) / 解像度: 3.4 Å 32898 7wyy EMDB-32898, PDB-7wyy: Cryo-EM structure of Na+,K+-ATPase in the E2P state formed by inorganic phosphate with istaroxime 手法: EM (単粒子) / 解像度: 3.9 Å 32899 7wyz EMDB-32899, PDB-7wyz: Cryo-EM structure of Na+,K+-ATPase in the E2P state formed by ATP with ouabain 手法: EM (単粒子) / 解像度: 3.4 Å 32900 7wz0 EMDB-32900, PDB-7wz0: Cryo-EM structure of Na+,K+-ATPase in the E2P state formed by inorganic phosphate with ouabain 手法: EM (単粒子) / 解像度: 3.0 Å PDB-7wys: Crystal structures of Na+,K+-ATPase in complex with istaroxime 手法: X-RAY DIFFRACTION / 解像度: 3.71 Å PDB-7wyt: Crystal structures of Na+,K+-ATPase in complex with ouabain 手法: X-RAY DIFFRACTION / 解像度: 2.9 Å
化合物	ChemComp-MG: Unknown entry / マグネシウムジカチオン ChemComp-NA: Unknown entry / ナトリウムカチオン ChemComp-PCW: 1,2-DIOLEOYL-SN-GLYCERO-3-PHOSPHOCHOLINE / O-(1-O,2-O-ジオレオイル-L-グリセロ-3-ホスホ)コリン / DOPC, リン脂質YM ChemComp-7Q2: (3E,5S,8R,9S,10R,13S,14S)-3-(2-azanylethoxyimino)-10,13-dimethyl-1,2,4,5,7,8,9,11,12,14,15,16-dodecahydrocyclopenta[a]phenanthrene-6,17-dione / イスタロキシム ChemComp-CLR: CHOLESTEROL / コレステロ-ル ChemComp-NAG: 2-acetamido-2-deoxy-beta-D-glucopyranose / N-アセチル-β-D-グルコサミン ChemComp-HOH: WATER ChemComp-OBN: OUABAIN / ウアバイン ChemComp-ATP: ADENOSINE-5'-TRIPHOSPHATE / ATP / ATP, エネルギー貯蔵分子YM
由来	squalus acanthias (アブラツノザメ) spiny dogfish (アブラツノザメ) sus scrofa (ブタ)
キーワード	MEMBRANE PROTEIN / Na+ / K+-ATPase / ion transport / Cardiotonic steroids / TRANSPORT PROTEIN