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	Rapid small-scale nanobody-assisted purification of ryanodine receptors for cryo-EM.
ジャーナル・号・ページ	J Biol Chem, Vol. 300, Issue 10, Page 107734, Year 2024
掲載日	2024年9月2日
著者	Chenyao Li / Katrien Willegems / Tomasz Uchański / Els Pardon / Jan Steyaert / Rouslan G Efremov /
PubMed 要旨	Ryanodine receptors (RyRs) are large Ca release channels residing in the endoplasmic or sarcoplasmic reticulum membrane. Three isoforms of RyRs have been identified in mammals, the disfunction of ...Ryanodine receptors (RyRs) are large Ca release channels residing in the endoplasmic or sarcoplasmic reticulum membrane. Three isoforms of RyRs have been identified in mammals, the disfunction of which has been associated with a series of life-threatening diseases. The need for large amounts of native tissue or eukaryotic cell cultures limits advances in structural studies of RyRs. Here, we report a method that utilizes nanobodies to purify RyRs from only 5 mg of total protein. The purification process, from isolated membranes to cryo-EM grade protein, is achieved within 4 h on the bench, yielding protein usable for cryo-EM analysis. This is demonstrated by solving the structures of rabbit RyR1, solubilized in detergent, reconstituted into lipid nanodiscs or liposomes, and bovine RyR2 reconstituted in nanodisc, and mouse RyR2 in detergent. The reported method facilitates structural studies of RyRs directed toward drug development and is useful in cases where the amount of starting material is limited.
リンク	J Biol Chem / PubMed:39233227 / PubMed Central
手法	EM (単粒子)
解像度	3.1 - 4.7 Å
構造データ	19463 8rrs EMDB-19463, PDB-8rrs: Structure of mouse RyR2 solubilised in detergent in open state in complex with Ca2+, ATP, caffeine and Nb9657. 手法: EM (単粒子) / 解像度: 3.4 Å 19464 8rrt EMDB-19464: Structure of rabbit RyR1 reconstituted into lipid liposomes in open state in complex with Ca2+, ATP, caffeine and Nb9657. PDB-8rrt: Structure of rabbit RyR1 reconstituted into lipid liposomes in open state in complex with FKBP and Nb9657 手法: EM (単粒子) / 解像度: 4.6 Å 19465 8rru EMDB-19465: Structure of RyR1 reconstituted into lipid liposomes in primed state in complex with Ca2+, ATP, caffeine and Nb9657. PDB-8rru: Structure of RyR1 reconstituted into lipid liposomes in primed state in complex with FKBP and Nb9657. 手法: EM (単粒子) / 解像度: 4.7 Å 19466 8rrv EMDB-19466, PDB-8rrv: Structure of RyR1 in detergent in close state in complex with FKBP and Nb9657. 手法: EM (単粒子) / 解像度: 3.2 Å 19467 8rrw EMDB-19467: Structure of RyR1 in detergent in open state in complex with Ca2+, ATP, caffeine and Nb9657. PDB-8rrw: Structure of RyR1 in detergent in open state in complex with FKBP and Nb9657. 手法: EM (単粒子) / 解像度: 4.2 Å 19468 8rrx EMDB-19468, PDB-8rrx: Structure of RyR1 reconstituted into lipid nanodisc in primed state in complex with Ca2+, ATP, caffeine and Nb9657 手法: EM (単粒子) / 解像度: 3.1 Å 19472 8rs0 EMDB-19472: Structure of RyR1 solubilised in detergent in primed state in complex with Ca2+, ATP, caffeine and Nb9657 PDB-8rs0: Structure of RyR1 in detergent in primed state in complex with nanobody and FKBP 手法: EM (単粒子) / 解像度: 3.3 Å
化合物	ChemComp-ATP: ADENOSINE-5'-TRIPHOSPHATE / ATP / ATP, エネルギー貯蔵分子YM ChemComp-CFF: CAFFEINE / カフェイン / 薬剤YM ChemComp-ZN: Unknown entry ChemComp-CA: Unknown entry / カルシウムジカチオン ChemComp-POV: (2S)-3-(hexadecanoyloxy)-2-[(9Z)-octadec-9-enoyloxy]propyl 2-(trimethylammonio)ethyl phosphate / リン脂質*YM
由来	mus musculus (ハツカネズミ) vicugna pacos (アルパカ) oryctolagus cuniculus (ウサギ)
キーワード	TRANSPORT PROTEIN / Ion channel / Ca2+ / tetramer