Activity-based profiling of cullin–RING E3 networks by conformation-specific probes
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Activity-based profiling of cullin–RING E3 networks by conformation-specific probes. / Henneberg, Lukas T.; Singh, Jaspal; Duda, David M.; Baek, Kheewoong; Yanishevski, David; Murray, Peter J.; Mann, Matthias; Sidhu, Sachdev S.; Schulman, Brenda A.
In: Nature Chemical Biology, Vol. 19, 2023, p. 1513-1523.Research output: Contribution to journal › Journal article › Research › peer-review
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TY - JOUR
T1 - Activity-based profiling of cullin–RING E3 networks by conformation-specific probes
AU - Henneberg, Lukas T.
AU - Singh, Jaspal
AU - Duda, David M.
AU - Baek, Kheewoong
AU - Yanishevski, David
AU - Murray, Peter J.
AU - Mann, Matthias
AU - Sidhu, Sachdev S.
AU - Schulman, Brenda A.
N1 - Publisher Copyright: © 2023, The Author(s).
PY - 2023
Y1 - 2023
N2 - 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. [Figure not available: see fulltext.]
AB - 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. [Figure not available: see fulltext.]
U2 - 10.1038/s41589-023-01392-5
DO - 10.1038/s41589-023-01392-5
M3 - Journal article
C2 - 37653169
AN - SCOPUS:85169157129
VL - 19
SP - 1513
EP - 1523
JO - Nature Chemical Biology
JF - Nature Chemical Biology
SN - 1552-4450
ER -
ID: 367909175