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Structure paper

タイトル	High-resolution structure determination of sub-100 kDa complexes using conventional cryo-EM.
ジャーナル・号・ページ	Nat Commun, Vol. 10, Issue 1, Page 1032, Year 2019
掲載日	2019年3月4日
著者	Mark A Herzik / Mengyu Wu / Gabriel C Lander /
PubMed 要旨	Determining high-resolution structures of biological macromolecules amassing less than 100 kilodaltons (kDa) has been a longstanding goal of the cryo-electron microscopy (cryo-EM) community. While ...Determining high-resolution structures of biological macromolecules amassing less than 100 kilodaltons (kDa) has been a longstanding goal of the cryo-electron microscopy (cryo-EM) community. While the Volta phase plate has enabled visualization of specimens in this size range, this instrumentation is not yet fully automated and can present technical challenges. Here, we show that conventional defocus-based cryo-EM methodologies can be used to determine high-resolution structures of specimens amassing less than 100 kDa using a transmission electron microscope operating at 200 keV coupled with a direct electron detector. Our ~2.7 Å structure of alcohol dehydrogenase (82 kDa) proves that bound ligands can be resolved with high fidelity to enable investigation of drug-target interactions. Our ~2.8 Å and ~3.2 Å structures of methemoglobin demonstrate that distinct conformational states can be identified within a dataset for proteins as small as 64 kDa. Furthermore, we provide the sub-nanometer cryo-EM structure of a sub-50 kDa protein.
リンク	Nat Commun / PubMed:30833564 / PubMed Central
手法	EM (単粒子)
解像度	2.8 - 6.0 Å
構造データ	0406 6nbb EMDB-0406: Single-particle cryo-EM reconstruction of horse liver alcohol dehydrogenase using 200 keV PDB-6nbb: Horse liver alcohol dehydrogenase determined using single-particle cryo-EM at 200 keV 手法: EM (単粒子) / 解像度: 2.9 Å 0407 6nbc EMDB-0407: Single-particle cryo-EM reconstruction of human methemoglobin using 200 keV, state 1 PDB-6nbc: human methemoglobin state 1 determined using single-particle cryo-EM at 200 keV 手法: EM (単粒子) / 解像度: 2.8 Å 0408 6nbd EMDB-0408: Single-particle cryo-EM reconstruction of human methemoglobin using 200 keV, state 2 PDB-6nbd: Human methemoglobin state 2 determined using single-particle cryo-EM at 200 keV 手法: EM (単粒子) / 解像度: 3.2 Å EMDB-0409: Single-particle cryo-EM reconstruction of the catalytic subunit of protein kinase A bound to ATP and IP20 using 200 keV 手法: EM (単粒子) / 解像度: 6.0 Å
化合物	ChemComp-NAD: NICOTINAMIDE-ADENINE-DINUCLEOTIDE / NADYM / ニコチンアミドアデニンジヌクレオチド ChemComp-ZN: Unknown entry ChemComp-HEM*: PROTOPORPHYRIN IX CONTAINING FE / Heme B
由来	equus caballus (ウマ) homo sapiens (ヒト) Mus musculus (ハツカネズミ)
キーワード	OXIDOREDUCTASE (酸化還元酵素) / dehydrogenase (脱水素酵素) / NADH-binding / homo-2-mer / OXYGEN TRANSPORT (血液) / heme-binding / hetero-4-mer / globin (グロビン)