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	Preparation of oxygen-sensitive proteins for high-resolution cryoEM structure determination using blot-free vitrification.
ジャーナル・号・ページ	Nat Commun, Vol. 16, Issue 1, Page 3528, Year 2025
掲載日	2025年4月14日
著者	Brian D Cook / Sarah M Narehood / Kelly L McGuire / Yizhou Li / F Akif Tezcan / Mark A Herzik /
PubMed 要旨	High-quality grid preparation for single-particle cryogenic electron microscopy (cryoEM) remains a bottleneck for routinely obtaining high-resolution structures. The issues that arise from ...High-quality grid preparation for single-particle cryogenic electron microscopy (cryoEM) remains a bottleneck for routinely obtaining high-resolution structures. The issues that arise from traditional grid preparation workflows are particularly exacerbated for oxygen-sensitive proteins, including metalloproteins, whereby oxygen-induced damage and alteration of oxidation states can result in protein inactivation, denaturation, and/or aggregation. Indeed, 99% of the current structures in the EMBD were prepared aerobically and limited successes for anaerobic cryoEM grid preparation exist. Current practices for anaerobic grid preparation involve a vitrification device located in an anoxic chamber, which presents significant challenges including temperature and humidity control, optimization of freezing conditions, costs for purchase and operation, as well as accessibility. Here, we present a streamlined approach that allows for the vitrification of oxygen-sensitive proteins in reduced states using an automated blot-free grid vitrification device - the SPT Labtech chameleon. This robust workflow allows for high-resolution structure determination of dynamic, oxygen-sensitive proteins, of varying complexity and molecular weight.
リンク	Nat Commun / PubMed:40229244 / PubMed Central
手法	EM (単粒子)
解像度	2.08 - 3.01 Å
構造データ	45815 9cqm EMDB-45815, PDB-9cqm: Human OxyHb (C1 symmetry) obtained using the SPT Labtech chameleon In the presence of 25 uM sodium dithionite under Al's oil 手法: EM (単粒子) / 解像度: 2.55 Å 45816 9cqn EMDB-45816, PDB-9cqn: Human OxyHb (C2 symmetry) obtained using the SPT Labtech chameleon In the presence of 25 uM sodium dithionite under Al's oil 手法: EM (単粒子) / 解像度: 2.37 Å 45817 9cqo EMDB-45817, PDB-9cqo: Human metHb (C1 symmetry) obtained using the SPT Labtech chameleon 手法: EM (単粒子) / 解像度: 3.01 Å 45818 9cqp EMDB-45818, PDB-9cqp: Human metHb (C2 symmetry) obtained using the SPT Labtech chameleon 手法: EM (単粒子) / 解像度: 2.78 Å 45819 9cqq EMDB-45819, PDB-9cqq: Human metHb (C1 symmetry) obtained using the SPT Labtech chameleon under Al's Oil 手法: EM (単粒子) / 解像度: 2.91 Å 45820 9cqr EMDB-45820, PDB-9cqr: Human metHb (C2 symmetry) obtained using the SPT Labtech chameleon under Al's Oil 手法: EM (単粒子) / 解像度: 2.7 Å 45821 9cqs EMDB-45821, PDB-9cqs: Human OxyHb (C1 symmetry) obtained using the SPT Labtech chameleon In the presence of 5 mM sodium dithionite under Al's oil 手法: EM (単粒子) / 解像度: 2.52 Å 45822 9cqt EMDB-45822, PDB-9cqt: Human OxyHb (C2 symmetry) obtained using the SPT Labtech chameleon In the presence of 5 mM sodium dithionite under Al's oil 手法: EM (単粒子) / 解像度: 2.37 Å 45823 9cqu EMDB-45823, PDB-9cqu: Human OxyHb (C1 symmetry) obtained using the SPT Labtech chameleon In the presence of 20 mM sodium dithionite under Al's oil 手法: EM (単粒子) / 解像度: 2.72 Å 45824 9cqv EMDB-45824, PDB-9cqv: Human DeoxyHb (C1 symmetry) obtained using the SPT Labtech chameleon In the presence of 60 mM sodium dithionite under Al's oil 手法: EM (単粒子) / 解像度: 2.75 Å 45825 9cqw EMDB-45825, PDB-9cqw: Human DeoxyHb (C2 symmetry) obtained using the SPT Labtech chameleon In the presence of 60 mM sodium dithionite under Al's oil 手法: EM (単粒子) / 解像度: 2.61 Å 45826 9cqx EMDB-45826, PDB-9cqx: Azotobacter vinelandii Oxidized MoFeP (C1 symmetry) obtained using the SPT Labtech chameleon 手法: EM (単粒子) / 解像度: 2.51 Å 45827 9cqy EMDB-45827, PDB-9cqy: Azotobacter vinelandii Oxidized MoFeP (C2 symmetry) obtained using the SPT Labtech chameleon 手法: EM (単粒子) / 解像度: 2.39 Å 45828 9cqz EMDB-45828, PDB-9cqz: Azotobacter vinelandii Reduced MoFeP (C1 symmetry) obtained using the SPT Labtech chameleon of 20 mM sodium dithionite under Al's oil 手法: EM (単粒子) / 解像度: 2.19 Å 45829 9cr0 EMDB-45829, PDB-9cr0: Azotobacter vinelandii Reduced MoFeP (C2 symmetry) obtained using the SPT Labtech chameleon of 20 mM sodium dithionite under Al's oil 手法: EM (単粒子) / 解像度: 2.08 Å 48381 9mly EMDB-48381, PDB-9mly: Azotobacter vinelandii Reduced MoFeP (C1 symmetry) obtained using the SPT Labtech chameleon of 5 mM sodium dithionite under Al's oil 手法: EM (単粒子) / 解像度: 2.33 Å 48382 9mlz EMDB-48382, PDB-9mlz: Azotobacter vinelandii Reduced MoFeP (C2 symmetry) obtained using the SPT Labtech chameleon of 5 mM sodium dithionite under Al's oil 手法: EM (単粒子) / 解像度: 2.19 Å 48383 9mm0 EMDB-48383, PDB-9mm0: Azotobacter vinelandii Reduced MoFeP (C1 symmetry) obtained using the SPT Labtech chameleon of 60 mM sodium dithionite under Al's oil 手法: EM (単粒子) / 解像度: 2.19 Å 48384 9mm1 EMDB-48384, PDB-9mm1: Azotobacter vinelandii Reduced MoFeP (C2 symmetry) obtained using the SPT Labtech chameleon of 60 mM sodium dithionite under Al's oil 手法: EM (単粒子) / 解像度: 2.08 Å
化合物	ChemComp-HEM: PROTOPORPHYRIN IX CONTAINING FE ChemComp-OXY: OXYGEN MOLECULE / ペルオキシド ChemComp-HOH: WATER ChemComp-HCA: 3-HYDROXY-3-CARBOXY-ADIPIC ACID / (2R)-2-ヒドロキシ-1,2,4-ブタントリカルボン酸 ChemComp-ICS: iron-sulfur-molybdenum cluster with interstitial carbon ChemComp-FE: Unknown entry ChemComp-CLF: FE(8)-S(7) CLUSTER
由来	homo sapiens (ヒト) azotobacter vinelandii (窒素固定)
キーワード	OXYGEN BINDING / Hemoglobin / anaerobic / oxygen / heme / METAL BINDING PROTEIN / Nitrogenase / FeMoCo / nitrogen / P-cluster