Activity-based profiling of cullin-RING E3 networks by conformation-specific probes.
Journal
Nature chemical biology
ISSN: 1552-4469
Titre abrégé: Nat Chem Biol
Pays: United States
ID NLM: 101231976
Informations de publication
Date de publication:
Dec 2023
Dec 2023
Historique:
received:
26
01
2023
accepted:
29
06
2023
medline:
27
11
2023
pubmed:
1
9
2023
entrez:
31
8
2023
Statut:
ppublish
Résumé
The cullin-RING ubiquitin ligase (CRL) network comprises over 300 unique complexes that switch from inactive to activated conformations upon site-specific cullin modification by the ubiquitin-like protein NEDD8. Assessing cellular repertoires of activated CRL complexes is critical for understanding eukaryotic regulation. However, probes surveying networks controlled by site-specific ubiquitin-like protein modifications are lacking. We developed a synthetic antibody recognizing the active conformation of NEDD8-linked cullins. Implementing the probe to profile cellular networks of activated CUL1-, CUL2-, CUL3- and CUL4-containing E3s revealed the complexes responding to stimuli. Profiling several cell types showed their baseline neddylated CRL repertoires vary, and prime efficiency of targeted protein degradation. Our probe also unveiled differential rewiring of CRL networks across distinct primary cell activation pathways. Thus, conformation-specific probes can permit nonenzymatic activity-based profiling across a system of numerous multiprotein complexes, which in the case of neddylated CRLs reveals widespread regulation and could facilitate the development of degrader drugs.
Identifiants
pubmed: 37653169
doi: 10.1038/s41589-023-01392-5
pii: 10.1038/s41589-023-01392-5
pmc: PMC10667097
doi:
Substances chimiques
Cullin Proteins
0
Ubiquitin-Protein Ligases
EC 2.3.2.27
Ubiquitin
0
Ubiquitins
0
NEDD8 Protein
0
Types de publication
Journal Article
Langues
eng
Sous-ensembles de citation
IM
Pagination
1513-1523Subventions
Organisme : NCI NIH HHS
ID : P30 CA021765
Pays : United States
Organisme : NIGMS NIH HHS
ID : R01 GM129325
Pays : United States
Commentaires et corrections
Type : UpdateOf
Informations de copyright
© 2023. The Author(s).
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