8CAF

N8C_Fab3b in complex with NEDD8-CUL1(WHB)

Summary for 8CAF

• Entry DOI: 10.2210/pdb8caf/pdb
• Descriptor: Fab Light Chain, Fab Heavy Chain, Cullin-1, ... (4 entities in total)
• Functional Keywords: nedd8, cullin-ring ligase, ubiquitin, antibody, cell cycle
• Biological source: Homo sapiens; More
• Total number of polymer chains: 8
• Total formula weight: 128531.14

Authors

Duda, D.M.,Yanishevski, D.,Henneberg, L.T.,Schulman, B.A. (deposition date: 2023-01-24, release date: 2023-09-13, Last modification date: 2024-11-13)

Primary citation

Henneberg, L.T.,Singh, J.,Duda, D.M.,Baek, K.,Yanishevski, D.,Murray, P.J.,Mann, M.,Sidhu, S.S.,Schulman, B.A.
Activity-based profiling of cullin-RING E3 networks by conformation-specific probes.
Nat.Chem.Biol., 19:1513-1523, 2023

PubMed Abstract: 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.

PubMed: 37653169
DOI: 10.1038/s41589-023-01392-5

Experimental method

X-RAY DIFFRACTION (2.66 Å)