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	Structural insights into the human PA28-20S proteasome enabled by efficient tagging and purification of endogenous proteins.
ジャーナル・号・ページ	Proc Natl Acad Sci U S A, Vol. 119, Issue 33, Page e2207200119, Year 2022
掲載日	2022年8月16日
著者	Jianhua Zhao / Suraj Makhija / Chenyu Zhou / Hanxiao Zhang / YongQiang Wang / Monita Muralidharan / Bo Huang / Yifan Cheng /
PubMed 要旨	The ability to produce folded and functional proteins is a necessity for structural biology and many other biological sciences. This task is particularly challenging for numerous biomedically ...The ability to produce folded and functional proteins is a necessity for structural biology and many other biological sciences. This task is particularly challenging for numerous biomedically important targets in human cells, including membrane proteins and large macromolecular assemblies, hampering mechanistic studies and drug development efforts. Here we describe a method combining CRISPR-Cas gene editing and fluorescence-activated cell sorting to rapidly tag and purify endogenous proteins in HEK cells for structural characterization. We applied this approach to study the human proteasome from HEK cells and rapidly determined cryogenic electron microscopy structures of major proteasomal complexes, including a high-resolution structure of intact human PA28αβ-20S. Our structures reveal that PA28 with a subunit stoichiometry of 3α/4β engages tightly with the 20S proteasome. Addition of a hydrophilic peptide shows that polypeptides entering through PA28 are held in the antechamber of 20S prior to degradation in the proteolytic chamber. This study provides critical insights into an important proteasome complex and demonstrates key methodologies for the tagging of proteins from endogenous sources.
リンク	Proc Natl Acad Sci U S A / PubMed:35858375 / PubMed Central
手法	EM (単粒子)
解像度	2.7 - 3.4 Å
構造データ	24275 7nan EMDB-24275, PDB-7nan: Human 20S proteasome core particle 手法: EM (単粒子) / 解像度: 2.8 Å 24276 7nao 8cxb EMDB-24276, PDB-7nao: Human PA28-20S proteasome complex PDB-8cxb: Human PA28-20S (PA28-4a3b) 手法: EM (単粒子) / 解像度: 2.9 Å 24277 7nap EMDB-24277, PDB-7nap: Human PA28-20S-PA28 proteasome complex 手法: EM (単粒子) / 解像度: 3.2 Å 24278 7naq EMDB-24278, PDB-7naq: Human PA200-20S proteasome complex 手法: EM (単粒子) / 解像度: 3.2 Å 27013 8cvr EMDB-27013, PDB-8cvr: Human 20S proteasome with MG-132 手法: EM (単粒子) / 解像度: 2.7 Å EMDB-27014: Human PA28-20S proteasome with MG-132 (mixed PA28 subunits) 手法: EM (単粒子) / 解像度: 2.8 Å 27015 8cvs EMDB-27015, PDB-8cvs: Human PA200-20S proteasome with MG-132 手法: EM (単粒子) / 解像度: 3.1 Å EMDB-27016: Human 19S-20S proteasome, state SA 手法: EM (単粒子) / 解像度: 3.3 Å EMDB-27017: Human 19S-20S proteasome, state SD1 手法: EM (単粒子) / 解像度: 3.4 Å 27018 8cvt EMDB-27018, PDB-8cvt: Human 19S-20S proteasome, state SD2 手法: EM (単粒子) / 解像度: 3.0 Å EMDB-27019: Human 19S-20S proteasome, state SD3/EC2 手法: EM (単粒子) / 解像度: 3.3 Å
化合物	ChemComp-IHP: INOSITOL HEXAKISPHOSPHATE / フィチン酸 ChemComp-LDZ: N-[(benzyloxy)carbonyl]-L-leucyl-N-[(2S)-4-methyl-1-oxopentan-2-yl]-L-leucinamide / MG-132 / 阻害剤YM ChemComp-ATP: ADENOSINE-5'-TRIPHOSPHATE / ATP / ATP, エネルギー貯蔵分子YM ChemComp-ZN: Unknown entry
由来	homo sapiens (ヒト) human (ヒト)
キーワード	HYDROLASE / proteasome / 20S / core particle / PA28 / PA200 / HYDROLASE/INHIBITOR / proteolysis / protein degradation / complex / inhibitor / MG-132 / MG132 / HYDROLASE-INHIBITOR complex