Abstract
Through targeting essential cellular regulators for ubiquitination and serving as a major platform for discovering proteolysis-targeting chimera (PROTAC) drugs, Cullin-2 (CUL2)-RING ubiquitin ligases (CRL2s) comprise an important family of CRLs. The founding members of CRLs, the CUL1-based CRL1s, are known to be activated by CAND1, which exchanges the variable substrate receptors associated with the common CUL1 core and promotes the dynamic assembly of CRL1s. Here we find that CAND1 inhibits CRL2-mediated protein degradation in human cells. This effect arises due to altered binding kinetics, involving CAND1 and CRL2VHL, as we illustrate that CAND1 dramatically increases the dissociation rate of CRL2s but barely accelerates the assembly of stable CRL2s. Using PROTACs that differently recruit neo-substrates to CRL2VHL, we demonstrate that the inhibitory effect of CAND1 helps distinguish target proteins with different affinities for CRL2s, presenting a mechanism for selective protein degradation with proper pacing in the changing cellular environment.
This is a preview of subscription content, access via your institution
Access options
Access Nature and 54 other Nature Portfolio journals
Get Nature+, our best-value online-access subscription
$29.99 / 30 days
cancel any time
Subscribe to this journal
Receive 12 print issues and online access
$189.00 per year
only $15.75 per issue
Buy this article
- Purchase on Springer Link
- Instant access to full article PDF
Prices may be subject to local taxes which are calculated during checkout
Similar content being viewed by others
Data availability
All data supporting the findings of this study are available within the paper. The proteomics data have been deposited to the PRIDE with the dataset identifier PXD045609. The UniProt Homo sapiens protein database is accessible through the link https://www.uniprot.org/proteomes/. Source data are provided with this paper.
References
Pohl, C. & Dikic, I. Cellular quality control by the ubiquitin-proteasome system and autophagy. Science 366, 818–822 (2019).
Harper, J. W. & Schulman, B. A. Cullin-RING ubiquitin ligase regulatory circuits: a quarter century beyond the F-box hypothesis. Annu. Rev. Biochem. 90, 403–429 (2021).
Lydeard, J. R., Schulman, B. A. & Harper, J. W. Building and remodelling Cullin-RING E3 ubiquitin ligases. EMBO Rep. 14, 1050–1061 (2013).
Zimmerman, E. S., Schulman, B. A. & Zheng, N. Structural assembly of cullin-RING ubiquitin ligase complexes. Curr. Opin. Struct. Biol. 20, 714–721 (2010).
Diaz, S., Wang, K., Sjogren, B. & Liu, X. Roles of Cullin-RING ubiquitin ligases in cardiovascular diseases. Biomolecules 12, 416 (2022).
Kamura, T., Conrad, M. N., Yan, Q., Conaway, R. C. & Conaway, J. W. The Rbx1 subunit of SCF and VHL E3 ubiquitin ligase activates Rub1 modification of cullins Cdc53 and Cul2. Genes Dev. 13, 2928–2933 (1999).
Kamura, T. et al. VHL-box and SOCS-box domains determine binding specificity for Cul2-Rbx1 and Cul5-Rbx2 modules of ubiquitin ligases. Genes Dev. 18, 3055–3065 (2004).
Mahrour, N. et al. Characterization of Cullin-box sequences that direct recruitment of Cul2-Rbx1 and Cul5-Rbx2 modules to Elongin BC-based ubiquitin ligases. J. Biol. Chem. 283, 8005–8013 (2008).
Sherpa, D., Chrustowicz, J. & Schulman, B. A. How the ends signal the end: regulation by E3 ubiquitin ligases recognizing protein termini. Mol. Cell 82, 1424–1438 (2022).
Maxwell, P. H. et al. The tumour suppressor protein VHL targets hypoxia-inducible factors for oxygen-dependent proteolysis. Nature 399, 271–275 (1999).
Liu, X., Zurlo, G. & Zhang, Q. The roles of Cullin-2 E3 ubiquitin ligase complex in cancer. Adv. Exp. Med Biol. 1217, 173–186 (2020).
Bekes, M., Langley, D. R. & Crews, C. M. PROTAC targeted protein degraders: the past is prologue. Nat. Rev. Drug Discov. 21, 181–200 (2022).
Cowan, A. D. & Ciulli, A. Driving E3 ligase substrate specificity for targeted protein degradation: lessons from nature and the laboratory. Annu. Rev. Biochem. 91, 295–319 (2022).
Baek, K., Scott, D. C. & Schulman, B. A. NEDD8 and ubiquitin ligation by cullin-RING E3 ligases. Curr. Opin. Struct. Biol. 67, 101–109 (2021).
Baek, K. et al. NEDD8 nucleates a multivalent cullin-RING-UBE2D ubiquitin ligation assembly. Nature 578, 461–466 (2020).
Deshaies, R. J. & Pierce, N. W. Transfer of ubiquitin protein caught in the act. Nature 578, 372–373 (2020).
Saha, A. & Deshaies, R. J. Multimodal activation of the ubiquitin ligase SCF by Nedd8 conjugation. Mol. Cell 32, 21–31 (2008).
Rozen, S. et al. CSNAP is a stoichiometric subunit of the COP9 signalosome. Cell Rep. 13, 585–598 (2015).
Gutierrez, C. et al. Structural dynamics of the human COP9 signalosome revealed by cross-linking mass spectrometry and integrative modeling. Proc. Natl Acad. Sci. USA 117, 4088–4098 (2020).
Schulze-Niemand, E. & Naumann, M. The COP9 signalosome: a versatile regulatory hub of Cullin-RING ligases. Trends Biochem. Sci. 48, 82–95 (2023).
Zhang, Y. et al. Adaptive exchange sustains cullin-RING ubiquitin ligase networks and proper licensing of DNA replication. Proc. Natl Acad. Sci. USA 119, e2205608119 (2022).
Pintard, L. et al. Neddylation and deneddylation of CUL-3 is required to target MEI-1/katanin for degradation at the meiosis-to-mitosis transition in C. elegans. Curr. Biol. 13, 911–921 (2003).
Wolf, D. A., Zhou, C. & Wee, S. The COP9 signalosome: an assembly and maintenance platform for cullin ubiquitin ligases? Nat. Cell Biol. 5, 1029–1033 (2003).
Cope, G. A. & Deshaies, R. J. COP9 signalosome: a multifunctional regulator of SCF and other cullin-based ubiquitin ligases. Cell 114, 663–671 (2003).
Zheng, J. et al. CAND1 binds to unneddylated CUL1 and regulates the formation of SCF ubiquitin E3 ligase complex. Mol. Cell 10, 1519–1526 (2002).
Liu, J., Furukawa, M., Matsumoto, T. & Xiong, Y. NEDD8 modification of CUL1 dissociates p120CAND1, an inhibitor of CUL1-SKP1 binding and SCF ligases. Mol. Cell 10, 1511–1518 (2002).
Goldenberg, S. J. et al. Structure of the Cand1-Cul1-Roc1 complex reveals regulatory mechanisms for the assembly of the multisubunit cullin-dependent ubiquitin ligases. Cell 119, 517–528 (2004).
Reitsma, J. M. et al. Composition and regulation of the cellular repertoire of SCF ubiquitin ligases. Cell 171, 1326–1339 e14 (2017).
Liu, X. et al. Cand1-mediated adaptive exchange mechanism enables variation in F-box protein expression. Mol. Cell 69, 773–786 e6 (2018).
Pierce, N. W. et al. Cand1 promotes assembly of new SCF complexes through dynamic exchange of F box proteins. Cell 153, 206–215 (2013).
Baek, K. et al. Systemwide disassembly and assembly of SCF ubiquitin ligase complexes. Cell 186, 1895–1911 e21 (2023).
Wang, K., Deshaies, R. J. & Liu, X. Assembly and regulation of CRL ubiquitin ligases. Adv. Exp. Med Biol. 1217, 33–46 (2020).
Li, L., Wang, K., Zhou, Y. & Liu, X. Review: a silent concert in developing plants: dynamic assembly of cullin-RING ubiquitin ligases. Plant Sci. 330, 111662 (2023).
Li, L. et al. CAND1 is required for pollen viability in Arabidopsis thaliana–a test of the adaptive exchange hypothesis. Front Plant Sci. 13, 866086 (2022).
Wu, S. et al. CAND1 controls in vivo dynamics of the cullin 1-RING ubiquitin ligase repertoire. Nat. Commun. 4, 1642 (2013).
Zemla, A. et al. CSN- and CAND1-dependent remodelling of the budding yeast SCF complex. Nat. Commun. 4, 1641 (2013).
Zhang, W. et al. Genetic analysis of CAND1-CUL1 interactions in Arabidopsis supports a role for CAND1-mediated cycling of the SCFTIR1 complex. Proc. Natl Acad. Sci. USA 105, 8470–8475 (2008).
Chuang, H. W., Zhang, W. & Gray, W. M. Arabidopsis ETA2, an apparent ortholog of the human cullin-interacting protein CAND1, is required for auxin responses mediated by the SCF(TIR1) ubiquitin ligase. Plant Cell 16, 1883–1897 (2004).
Feng, S. et al. Arabidopsis CAND1, an unmodified CUL1-interacting protein, is involved in multiple developmental pathways controlled by ubiquitin/proteasome-mediated protein degradation. Plant Cell 16, 1870–1882 (2004).
Rusnac, D. V. & Zheng, N. Structural biology of CRL ubiquitin ligases. Adv. Exp. Med. Biol. 1217, 9–31 (2020).
Lo, S. C. & Hannink, M. CAND1-mediated substrate adaptor recycling is required for efficient repression of Nrf2 by Keap1. Mol. Cell. Biol. 26, 1235–1244 (2006).
Reichermeier, K. M. et al. PIKES analysis reveals response to degraders and key regulatory mechanisms of the CRL4 network. Mol. Cell 77, 1092–1106 e9 (2020).
Mayor-Ruiz, C. et al. Plasticity of the Cullin-RING ligase repertoire shapes sensitivity to ligand-induced protein degradation. Mol. Cell 75, 849–858 e8 (2019).
Sievers, Q. L., Gasser, J. A., Cowley, G. S., Fischer, E. S. & Ebert, B. L. Genome-wide screen identifies cullin-RING ligase machinery required for lenalidomide-dependent CRL4(CRBN) activity. Blood 132, 1293–1303 (2018).
Soucy, T. A. et al. An inhibitor of NEDD8-activating enzyme as a new approach to treat cancer. Nature 458, 732–736 (2009).
Diaz, S., Li, L., Wang, K. & Liu, X. Expression and purification of functional recombinant CUL2*RBX1 from E. coli. Sci. Rep. 11, 11224 (2021).
Raina, K. et al. PROTAC-induced BET protein degradation as a therapy for castration-resistant prostate cancer. Proc. Natl Acad. Sci. USA 113, 7124–7129 (2016).
Wang, K., Reichermeier, K. M. & Liu, X. Quantitative analyses for effects of neddylation on CRL2(VHL) substrate ubiquitination and degradation. Protein Sci. 30, 2338–2345 (2021).
Koren, I. et al. The eukaryotic proteome is shaped by E3 ubiquitin ligases targeting C-terminal degrons. Cell 173, 1622–1635 e14 (2018).
Yeh, C. W. et al. The C-degron pathway eliminates mislocalized proteins and products of deubiquitinating enzymes. EMBO J. 40, e105846 (2021).
Schlierf, A. et al. Targeted inhibition of the COP9 signalosome for treatment of cancer. Nat. Commun. 7, 13166 (2016).
Cope, G. A. et al. Role of predicted metalloprotease motif of Jab1/Csn5 in cleavage of Nedd8 from Cul1. Science 298, 608–611 (2002).
Schwechheimer, C. et al. Interactions of the COP9 signalosome with the E3 ubiquitin ligase SCFTIRI in mediating auxin response. Science 292, 1379–1382 (2001).
Nguyen, H. C., Yang, H., Fribourgh, J. L., Wolfe, L. S. & Xiong, Y. Insights into Cullin-RING E3 ubiquitin ligase recruitment: structure of the VHL-EloBC-Cul2 complex. Structure 23, 441–449 (2015).
Cardote, T. A. F., Gadd, M. S. & Ciulli, A. Crystal structure of the Cul2-Rbx1-EloBC-VHL ubiquitin ligase complex. Structure 25, 901–911 e3 (2017).
Mosadeghi, R. et al. Structural and kinetic analysis of the COP9-Signalosome activation and the cullin-RING ubiquitin ligase deneddylation cycle. eLife 5, e12102 (2016).
Faull, S. V. et al. Structural basis of Cullin 2 RING E3 ligase regulation by the COP9 signalosome. Nat. Commun. 10, 3814 (2019).
Shaaban, M. et al. Structural and mechanistic insights into the CAND1-mediated SCF substrate receptor exchange. Mol. Cell 83, 2332–2346 e8 (2023).
Roy, M. J. et al. SPR-measured dissociation kinetics of PROTAC ternary complexes influence target degradation rate. ACS Chem. Biol. 14, 361–368 (2019).
Gadd, M. S. et al. Structural basis of PROTAC cooperative recognition for selective protein degradation. Nat. Chem. Biol. 13, 514–521 (2017).
Imaide, S. et al. Trivalent PROTACs enhance protein degradation via combined avidity and cooperativity. Nat. Chem. Biol. 17, 1157–1167 (2021).
Lin, H. C. et al. CRL2 aids elimination of truncated selenoproteins produced by failed UGA/Sec decoding. Science 349, 91–95 (2015).
Lin, H. C. et al. C-terminal end-directed protein elimination by CRL2 ubiquitin ligases. Mol. Cell 70, 602–613 e3 (2018).
Timms, R. T. et al. A glycine-specific N-degron pathway mediates the quality control of protein N-myristoylation. Science 365, eaaw4912 (2019).
Fischer, E. S. et al. The molecular basis of CRL4DDB2/CSA ubiquitin ligase architecture, targeting, and activation. Cell 147, 1024–1039 (2011).
Wang, K. & Liu, X. Determining the effects of neddylation on Cullin-ring ligase-dependent protein ubiquitination. Curr. Protoc. 2, e401 (2022).
Garsamo, M., Zhou, Y. & Liu, X. Using in vitro fluorescence resonance energy transfer to study the dynamics of protein complexes at a millisecond time scale. J. Vis. Exp. 145, e59038 (2019).
Acknowledgements
We thank S.-O. Shan (California Institute of Technology) for helpful discussion on kinetics data. We thank S.J. Elledge (Harvard Medical School) and D.C. Scott (St. Jude Children’s Research Hospital) for reagents. Mass spectrometry was provided by the Indiana University School of Medicine Center for Proteome Analysis. The Octet RED384 system for BLI assays was provided by the Chemical Genomics Facility at Purdue Institute for Drug Discovery. This work was supported by National Institutes of Health grant no. R35GM138016 (to X.L.) and an American Heart Association Career Development Award no. 20CDA35270030 (to X.L.). The funders had no role in study design, data collection and analysis, decision to publish or preparation of the manuscript.
Author information
Authors and Affiliations
Contributions
K.W. and X.L. designed the experiments and prepared the manuscript. K.W. performed all experiments unless stated otherwise. S.D. performed the assay in Fig. 1b. L.L. generated baculovirus stock for protein expression and aided experiments in Extended Data Fig. 6b,c. J.R.L. synthesized GGGGKAMCA peptides, aided data interpretation and manuscript writing. X.L conceived the study and supervised the project.
Corresponding author
Ethics declarations
Competing interests
The authors declare no competing interests.
Peer review
Peer review information
Nature Structural & Molecular Biology thanks Alessio Ciulli and the other, anonymous, reviewer(s) for their contribution to the peer review of this work. Peer reviewer reports are available. Dimitris Typas was the primary editor on this article and managed its editorial process and peer review in collaboration with the rest of the editorial team. Peer reviewer reports are available.
Additional information
Publisher’s note Springer Nature remains neutral with regard to jurisdictional claims in published maps and institutional affiliations.
Extended data
Extended Data Fig. 1 ARV-771 induced degradation of BRD2 in the presence of elevated levels of VHL.
a-c, WT and DKO cells expressing the same level of VHL show no difference in BRD2 degradation. a, WT cells contained a lower level of VHL than the DKO cells. WTVHL (WT cells containing transgenic VHL) and DKO cells expressed equal levels of VHL. b, Cells in a were treated with 50 nM ARV-771 for the indicated time and immunoblotted with anti-BRD2 antibody. Quantitative analyses of BRD2 degradation were shown in c. Error bars represent SEM; n = 3 independent experiments.
Extended Data Fig. 2 Effects of CSN5i-3 on the degradation of CODD in the WT and DKO cells.
a, b, WT cells expressing FLAGCODD were pretreated with 200 µM desferrioxamine. DMSO or 1 µM CSN5i-3 was added to the medium 1 h before washing off the inhibitors. Cells were then treated with 60 μg/ml cycloheximide (CHX) for the indicated time and were immunoblotted with anti-FLAG antibody. Quantitative analyses of FLAGCODD degradation in a were shown in b. Error bars represent SEM; n = 3 independent experiments. c, d, Same as in a, b except that DKO cells were used. Error bars represent SEM; n = 3 independent experiments.
Extended Data Fig. 3 Assessing dynamic assembly and disassembly of CRL2s in human cell lysates.
a, Trial of bait protein immobilization for the BLI experiments. GSTVBC at various concentrations were incubated with anti-GST antibody coated biosensors, and the sensor response levels were recorded over time. For all BLI experiments in this study, 100 nM GSTVBC was used for loading, and a minimal response level of 1 nm was achieved before the kinetic binding assay was initiated. b, Representative results of GSTVBC loading for the BLI assay shown in Fig. 3a. c, Schematic illustration of CRISPR/Cas9-mediated epitope tagging on the endogenous CUL2 locus in DKO cells. Single-stranded oligodeoxynucleotide (ssODN) carrying the coding sequence for the StrepII tag was used as the homology-directed repair (HDR) template. Red arrows indicate primers (1 F, 2 F, 3 F, 1 R, 2 R) used for screening the correctly tagged cells. d, The guide RNA (gRNA) targeting CUL2 as illustrated in c is efficient in inducing double strain breaks at the target site. Cell populations electroporated with the plasmid expressing gRNA and Cas9 were harvested for genomic DNA extraction and PCR with the 3 F and 1 R primer set as illustrated in c. Sequencing chromatograms of PCR products spanning the target sites were shown. e, Successful gene editing in the knock-in clonal cell line was identified using the primer sets ‘1 F + 1 R’ and ‘2 F + 2 R’ as illustrated in c. f, Sequencing chromatograms confirming the correct genomic sequences at the 5’ and 3’ junctions of the HDR region in the knock-in cell line. g, Immunoblots validating the successful preparation of SILAC samples illustrated in Fig. 3d. h, Estimation of the VHL concentration in HEK293 cells. Total cell extract from WT cells and recombinant VHL standards were immunoblotted with anti-VHL antibody. Levels of VHL in the WT cells were calculated based on the standard curve generated from the known amounts of recombinant VHL standards.
Extended Data Fig. 4 Quantitative analyses of interactions between CUL2 and VBC, and between CUL2 and CAND1.
a, Sequence alignment of ELOC1-40 from different species. b, Representative results of GSTVBC loading for the BLI assay shown in Fig. 4a. c, d, BLI analyses revealing kon (c) and koff (d) for CUL2 binding to VBC that contains different ELOC isoforms. n = 3. See the ‘Methods’ section for more details. e, Measurements of kobs for VBC binding to CUL2. Real-time CFP fluorescence upon mixing increasing concentrations of VBCFlAsH to 10 nM CFPCUL2 in a stopped-flow fluorimeter. Signal changes with background (determined by the ‘buffer’ group) subtraction were fit to two-phase exponential curves. The fast-phase and slow-phase rates (kobs) were plotted to give kon,fast and kon,slow, respectively. f, g, Estimation of the CUL2•CAND1 dissociation rate in human cell lysate. f, Schematic workflow of the assay. Relative levels of co-precipitated CAND1HA were calculated from CAND1HA/StrepIICUL2 intensity ratios followed by normalizing these ratios to that obtained from the 0-min sample. The normalized data were fit to a single exponential curve to obtain t1/2 and koff (g). n = 2. h, i, CAND1 produced from insect cells, but not bacterial cells, forms CUL2•CAND1 as stable as the CUL2•CAND1 in human cell lysate (f,g,). h, GST pulldown analyses of recombinant CUL2 bound to GSTCAND1 produced from E. coli or insect cells in the presence of CUL2split chase for indicated time. Samples were fractionated on SDS-PAGE gels and immunoblotted with indicated antibodies. Intensity ratios of the CUL2 and GSTCAND1 bands were fit to single exponential curves to obtain t1/2 (i). n = 3. j, Measurement of kobs for CAND1 binding to CUL2. Real-time AMCA fluorescence upon addition of increasing concentrations of FlAsHCAND1 to CUL2AMCA. Signal changes were fit to one phase exponential curves. CAND1 was purified from insect cells. Error bars represent SEM (d) or SD (i). ‘n’ indicates number of independent experiments. P values were determined by two-sided t-tests with no adjustments.
Extended Data Fig. 5 Interactions between CUL2 and VBC in the presence of CAND1.
a, Measurements of kobs for VBC binding to CUL2 preassembled with CAND1. Same as in Extended Data Fig. 4e except that 20 nM CAND1 was preincubated with 10 nM CFPCUL2. Signal changes with background subtraction (background determined by the ‘buffer’ group) were fit to one-phase exponential curves to give kobs. CAND1 was produced from insect cells. b,c, CAND1 accelerates the disassembly of CRL2VHL in human cell lysate. The schematic workflow of the experiment is depicted in b. Samples were immunoblotted with anti-VHL, anti-StrepII, and anti-CAND1 antibodies (c). Both short (S.E.) and long (L.E.) exposure for anti-VHL signals are shown. d, CAND1 accelerates the disassembly of CUL2•VBC in vitro. Same as in Fig. 5c except with 100 nM of different CAND1 proteins added in the presence of VBC chase. Both CAND1 and CAND1β++ were generated from bacterial cells. CAND1β++ showed no notable effect on the CFPCUL2-VBCFlAsH FRET signal. e, Structural alignment of CUL1•CAND1 complex (PDB ID: 1U6G) and CUL2•VBC complex (PDB ID: 5N4W) based on the N-terminal amino acid sequences (1-300) of cullins. f, Measurements of kobs for CUL2•VBC dissociation by CAND1. Real-time detection of CFP fluorescence in a stopped-flow apparatus upon addition of increasing concentrations of CAND1 together with 100 nM VBC chase to 10 nM CFPCUL2 preincubated with 10 nM VBCFlAsH. Final concentrations after mixing the two solutions in the stopped-flow fluorimeter were shown. Signal changes were fit to single exponential curves to obtain the kobs. CAND1 was produced from insect cells.
Extended Data Fig. 6 CUL2•CAND1 dissociation and its effect on CUL2 neddylation.
a, Measurements of kobs for FlAsHCAND1 binding to CUL2AMCA preassembled with VBC. Same as in Extended Data Fig. 4j except that 20 nM VBC was preincubated with 5 nM CUL2AMCA. Signal changes were fit to one phase exponential curves. CAND1 was purified from insect cells. b,c, kon for CAND1 binding to CUL2 preassembled with VBC. b, Measurement of kobs for FlAsHCAND1 binding to CUL2AMCA preassembled with VBC. Same as in Extended Data Fig. 6a except that 1 μM VBC was preincubated with 5 nM CUL2AMCA. Signal changes were fit to one-phase exponential curves to obtain kobs, which were plotted against FlAsHCAND1 concentrations in (c). Linear slope in (c) gave the kon. Error bars represent SEM; n = 4 independent experiments. CAND1 was purified from insect cells. d, HEK293 cells were treated with 1 µM CSN5i-3 for indicated time, and the rate of CUL2 neddylation was monitored using immunoblot with anti-CUL2 antibody. The proportion of unneddylated CUL2 in the whole CUL2 population (CUL2Total) versus time was plotted and fit to a single exponential curve to obtain the kobs. Error bars represent SEM; n = 3 independent experiments. CUL2N8: neddylated CUL2.
Extended Data Fig. 7 ZBC hardly accelerates the disassembly of CUL2•CAND1.
a, StrepII-tag affinity pulldown assay confirming the binding between recombinant StrepIICUL2 and ZBC. b, c, Similar to Fig. 6h, i except that VBC was replaced by ZBC. Samples were immunoblotted with anti-CAND1, anti-StrepII, and anti-ELOB antibodies.
Extended Data Fig. 8 PROTAC induced degradation of BRD2 and BRD3 in WT and DKO cells.
a, Schematic illustration for the MZ-1 and SIM1 mediated interactions between VHL and bromodomain proteins. b, As in Fig. 7a except that 0.3 µM MZ-1 was used in place of 1 µM MZ-1. Error bars represent SEM; n = 3 independent experiments.
Supplementary information
Source data
Source Data Fig. 1
Unprocessed gels and blots.
Source Data Fig. 1
Statistical source data.
Source Data Fig. 2
Unprocessed blots.
Source Data Fig. 2
Statistical source data.
Source Data Fig. 3
Unprocessed blots.
Source Data Fig. 3
Statistical source data.
Source Data Fig. 4
Unprocessed blots.
Source Data Fig. 4
Statistical source data.
Source Data Fig. 5
Statistical source data.
Source Data Fig. 6
Unprocessed blots.
Source Data Fig. 6
Statistical source data.
Source Data Fig. 7
Unprocessed blots.
Source Data Fig. 7
Statistical source data.
Source Data Extended Data Fig. 1
Unprocessed blots.
Source Data Extended Data Fig. 1
Statistical source data.
Source Data Extended Data Fig. 2
Unprocessed blots.
Source Data Extended Data Fig. 2
Statistical source data.
Source Data Extended Data Fig. 3
Unprocessed gels and blots.
Source Data Extended Data Fig. 3
Statistical source data.
Source Data Extended Data Fig. 4
Unprocessed blots.
Source Data Extended Data Fig. 4
Statistical source data.
Source Data Extended Data Fig. 5
Unprocessed blots.
Source Data Extended Data Fig. 5
Statistical source data.
Source Data Extended Data Fig. 6
Unprocessed blots.
Source Data Extended Data Fig. 6
Statistical source data.
Source Data Extended Data Fig. 7
Unprocessed gels and blots.
Source Data Extended Data Fig. 8
Unprocessed gels and blots.
Source Data Extended Data Fig. 8
Statistical source data.
Rights and permissions
Springer Nature or its licensor (e.g. a society or other partner) holds exclusive rights to this article under a publishing agreement with the author(s) or other rightsholder(s); author self-archiving of the accepted manuscript version of this article is solely governed by the terms of such publishing agreement and applicable law.
About this article
Cite this article
Wang, K., Diaz, S., Li, L. et al. CAND1 inhibits Cullin-2-RING ubiquitin ligases for enhanced substrate specificity. Nat Struct Mol Biol 31, 323–335 (2024). https://doi.org/10.1038/s41594-023-01167-5
Received:
Accepted:
Published:
Issue Date:
DOI: https://doi.org/10.1038/s41594-023-01167-5
