Characterization of Fully Recombinant Human 20S and 20S-PA200 Proteasome Complexes.
Allosteric Regulation
Animals
Binding Sites
Humans
Inositol Phosphates
/ metabolism
Models, Molecular
Nuclear Proteins
/ genetics
Proteasome Endopeptidase Complex
/ genetics
Protein Binding
Protein Conformation
Proteolysis
Recombinant Proteins
/ metabolism
Sf9 Cells
Spodoptera
Structure-Activity Relationship
20S-PA200
PA200
atomic model
cryo-EM
human
proteasome
recombinant
regulator
structure
Journal
Molecular cell
ISSN: 1097-4164
Titre abrégé: Mol Cell
Pays: United States
ID NLM: 9802571
Informations de publication
Date de publication:
03 10 2019
03 10 2019
Historique:
received:
28
01
2019
revised:
27
05
2019
accepted:
10
07
2019
pubmed:
2
9
2019
medline:
20
2
2020
entrez:
2
9
2019
Statut:
ppublish
Résumé
Proteasomes are essential in all eukaryotic cells. However, their function and regulation remain considerably elusive, particularly those of less abundant variants. We demonstrate the human 20S proteasome recombinant assembly and confirmed the recombinant complex integrity biochemically and with a 2.6 Å resolution cryo-EM map. To assess its competence to form higher-order assemblies, we prepared and analyzed recombinant human 20S-PA200, a poorly characterized nuclear complex. Its 3.0 Å resolution cryo-EM structure reveals the PA200 unique architecture; the details of its intricate interactions with the proteasome, resulting in unparalleled proteasome α ring rearrangements; and the molecular basis for PA200 allosteric modulation of the proteasome active sites. Non-protein cryo-EM densities could be assigned to PA200-bound inositol phosphates, and we speculate regarding their functional role. Here we open extensive opportunities to study the fundamental properties of the diverse and distinct eukaryotic proteasome variants and to improve proteasome targeting under different therapeutic conditions.
Identifiants
pubmed: 31473102
pii: S1097-2765(19)30544-1
doi: 10.1016/j.molcel.2019.07.014
pmc: PMC6863390
pii:
doi:
Substances chimiques
Inositol Phosphates
0
Nuclear Proteins
0
Recombinant Proteins
0
proteasome activator 200 kDa, human
0
Proteasome Endopeptidase Complex
EC 3.4.25.1
Types de publication
Journal Article
Research Support, Non-U.S. Gov't
Langues
eng
Sous-ensembles de citation
IM
Pagination
138-147.e5Subventions
Organisme : Medical Research Council
ID : MC_UP_1201/5
Pays : United Kingdom
Informations de copyright
Copyright © 2019 The Authors. Published by Elsevier Inc. All rights reserved.
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