Database of articles cited by EMDB/PDB/SASBDB data

Keywords
Structure methods	All X-ray diffraction NMR electron microscopy small angle scattering neutron diffraction fiber diffraction powder diffraction infrared spectroscopy EPR fluorescence transfer theoretical model Hybrid
Author
Journal
IF	>30 >20 >10 >5 all

Title	Structural insights into the human PA28-20S proteasome enabled by efficient tagging and purification of endogenous proteins.
Journal, issue, pages	Proc Natl Acad Sci U S A, Vol. 119, Issue 33, Page e2207200119, Year 2022
Publish date	Aug 16, 2022
Authors	Jianhua Zhao / Suraj Makhija / Chenyu Zhou / Hanxiao Zhang / YongQiang Wang / Monita Muralidharan / Bo Huang / Yifan Cheng /
PubMed Abstract	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.
External links	Proc Natl Acad Sci U S A / PubMed:35858375 / PubMed Central
Methods	EM (single particle)
Resolution	2.7 - 3.4 Å
Structure data	24275 7nan EMDB-24275, PDB-7nan: Human 20S proteasome core particle Method: EM (single particle) / Resolution: 2.8 Å 24276 7nao 8cxb EMDB-24276, PDB-7nao: Human PA28-20S proteasome complex PDB-8cxb: Human PA28-20S (PA28-4a3b) Method: EM (single particle) / Resolution: 2.9 Å 24277 7nap EMDB-24277, PDB-7nap: Human PA28-20S-PA28 proteasome complex Method: EM (single particle) / Resolution: 3.2 Å 24278 7naq EMDB-24278, PDB-7naq: Human PA200-20S proteasome complex Method: EM (single particle) / Resolution: 3.2 Å 27013 8cvr EMDB-27013, PDB-8cvr: Human 20S proteasome with MG-132 Method: EM (single particle) / Resolution: 2.7 Å EMDB-27014: Human PA28-20S proteasome with MG-132 (mixed PA28 subunits) Method: EM (single particle) / Resolution: 2.8 Å 27015 8cvs EMDB-27015, PDB-8cvs: Human PA200-20S proteasome with MG-132 Method: EM (single particle) / Resolution: 3.1 Å EMDB-27016: Human 19S-20S proteasome, state SA Method: EM (single particle) / Resolution: 3.3 Å EMDB-27017: Human 19S-20S proteasome, state SD1 Method: EM (single particle) / Resolution: 3.4 Å 27018 8cvt EMDB-27018, PDB-8cvt: Human 19S-20S proteasome, state SD2 Method: EM (single particle) / Resolution: 3.0 Å EMDB-27019: Human 19S-20S proteasome, state SD3/EC2 Method: EM (single particle) / Resolution: 3.3 Å
Chemicals	ChemComp-IHP: INOSITOL HEXAKISPHOSPHATE / Phytic acid ChemComp-LDZ: N-[(benzyloxy)carbonyl]-L-leucyl-N-[(2S)-4-methyl-1-oxopentan-2-yl]-L-leucinamide / inhibitorYM / MG132 ChemComp-ATP: ADENOSINE-5'-TRIPHOSPHATE / ATP, energy-carrying moleculeYM / Adenosine triphosphate ChemComp-ZN: Unknown entry
Source	homo sapiens (human) human (human)
Keywords	HYDROLASE / proteasome / 20S / core particle / PA28 / PA200 / HYDROLASE/INHIBITOR / proteolysis / protein degradation / complex / inhibitor / MG-132 / MG132 / HYDROLASE-INHIBITOR complex