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Cited by 158Protein Degradation and InhibitorsUbiquitin and proteasome pathwaysCell death mechanisms and regulation

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Development of a BCL-xL and BCL-2 dual degrader with improved anti-leukemic activity,

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Lv, Dongwen, Pal, Pratik, Liu, Xingui, Jia, Yannan, Thummuri, Dinesh, Zhang, Peiyi, Hu, Wanyi, Pei, Jing, Zhang, Qi, Zhou, Shuo, Khan, Sajid, Zhang, Xuan, Hua, Nan, Yang, Qingping, Arango, Sebastian, Zhang, Weizhou, Nayak, Digant, Olsen, Shaun K., Weintraub, Susan T., Hromas, Robert, Konopleva, Marina, Yuan, Yaxia, Zheng, Guangrong, Zhou, Daohong·2021·Nature Communications
DOI 10.1038/s41467-021-27210-xPMC PMC8617031
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Abstract

PROteolysis-TArgeting Chimeras (PROTACs) have emerged as an innovative drug development platform. However, most PROTACs have been generated empirically because many determinants of PROTAC specificity and activity remain elusive. Through computational modelling of the entire NEDD8-VHL Cullin RING E3 ubiquitin ligase (CRL VHL )/PROTAC/BCL-xL/UbcH5B(E2)-Ub/RBX1 complex, we find that this complex can only ubiquitinate the lysines in a defined band region on BCL-xL. Using this approach to guide our development of a series of ABT263-derived and VHL-recruiting PROTACs, we generate a potent BCL-xL and BCL-2 (BCL-xL/2) dual degrader with significantly improved antitumor activity against BCL-xL/2-dependent leukemia cells. Our results provide experimental evidence that the accessibility of lysines on a target protein plays an important role in determining the selectivity and potency of a PROTAC in inducing protein degradation, which may serve as a conceptual framework to guide the future development of PROTACs.

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Figures

Fig. 1 A band region of accessible lysines on BCL-xL. a A theoretical model of ubiquitination reaction. The protein of interest (POI), ubiquitin, and E2 enzyme are colored in green, pink, and blue, respectively. The conserved E2 serine/aspartate (CES/D) site, catalytic cysteine, and target lysine are showed in orange balls. The target lysine is firstly attracted to the CES/D site by electrostatic interaction, and subsequently attack the thioester bond between ubiquitin and E2 enzyme. b Atomic details of the catalytic mechanism. c The structure of DT2216. d A predicted band region of E2-accessible lysines on BCL-xL. The six selected lysine residues or lysine point mutation sites around BCL-xL are shown in sphere style. R34K, R102K, R132K, and K87 are in the predicted band region, and K20, K157, and G186K are outside of the band region. e A computational model of the entire CRLVHL/DT2216/BCL-xL/E2-Ub/RBX1 complex. The possible motion region of E2 is shown with dashed sector. The catalytic site motion region is represented by two blue dashed lines. The accessible region of the ubiquitination is represented by the two blue lines along the direction of E2 proximity to POI represented by an orange dashed arrow. The potential direction of swing of Cul2 and RBX1 are shown by a gray double-headed arrow. f The point mutation analyses confirm the existence of a band region of lysines on BCL-xL. WT: wild type Flag-BCL-xL; K-ko: all the surface lysines on BCL-xL are mutated to arginines; K157-only: all the surface lysines on BCL-xL are mutated to arginines except K157; K-ko-G186K, K-ko-R34K, K-ko-R102K, and K-ko-R132K represent that all the surface lysines on BCL-xL are mutated to arginines and then Gly186, Arg34, Arg102, and Arg132 are respectively mutated to lysine. β-actin was used as an equal loading control. The quantification of the relative Flag-BCL-xL protein content in the immunoblots is presented as mean values ± s.e.m. (n = 3 biologically independent experiments) in the bar graph (bottom panel). Statistical significance was calculated with unpaired two-tailed Student’s t-tests comparing DMSO- to DT2216-treated samples. **P < 0.01; P values were 0.0089, 0.0023, 0.0077, and 0.0026 for WT, K-ko-R34K, K-ko-R102K, and K-ko-R132K, respectively. Source data are provided as a Source data file.
Fig. 1 A band region of accessible lysines on BCL-xL. a A theoretical model of ubiquitination reaction. The protein of interest (POI), ubiquitin, and E2 enzyme are colored in green, pink, and blue, respectively. The conserved E2 serine/aspartate (CES/D) site, catalytic cysteine, and target lysine are showed in orange balls. The target lysine is firstly attracted to the CES/D site by electrostatic interaction, and subsequently attack the thioester bond between ubiquitin and E2 enzyme. b Atomic details of the catalytic mechanism. c The structure of DT2216. d A predicted band region of E2-accessible lysines on BCL-xL. The six selected lysine residues or lysine point mutation sites around BCL-xL are shown in sphere style. R34K, R102K, R132K, and K87 are in the predicted band region, and K20, K157, and G186K are outside of the band region. e A computational model of the entire CRLVHL/DT2216/BCL-xL/E2-Ub/RBX1 complex. The possible motion region of E2 is shown with dashed sector. The catalytic site motion region is represented by two blue dashed lines. The accessible region of the ubiquitination is represented by the two blue lines along the direction of E2 proximity to POI represented by an orange dashed arrow. The potential direction of swing of Cul2 and RBX1 are shown by a gray double-headed arrow. f The point mutation analyses confirm the existence of a band region of lysines on BCL-xL. WT: wild type Flag-BCL-xL; K-ko: all the surface lysines on BCL-xL are mutated to arginines; K157-only: all the surface lysines on BCL-xL are mutated to arginines except K157; K-ko-G186K, K-ko-R34K, K-ko-R102K, and K-ko-R132K represent that all the surface lysines on BCL-xL are mutated to arginines and then Gly186, Arg34, Arg102, and Arg132 are respectively mutated to lysine. β-actin was used as an equal loading control. The quantification of the relative Flag-BCL-xL protein content in the immunoblots is presented as mean values ± s.e.m. (n = 3 biologically independent experiments) in the bar graph (bottom panel). Statistical significance was calculated with unpaired two-tailed Student’s t-tests comparing DMSO- to DT2216-treated samples. **P < 0.01; P values were 0.0089, 0.0023, 0.0077, and 0.0026 for WT, K-ko-R34K, K-ko-R102K, and K-ko-R132K, respectively. Source data are provided as a Source data file.CC BY · PubMed Central
Fig. 2 Synthesis and evaluation of a series of BCL-xL and/or BCL-2 degraders. a The structure of the BCL-xL and/or BCL-2 degraders (racemic mixture). b The BCL-xL and BCL-2 degradation profiling of the PROTACs evaluated in 293T cells after the cells were treated with four different concentrations of indicated PROTACs for 16 h. The DC50 and Dmax were calculated using Prism based on the ImageJ-quantified western blots in Supplementary Fig. 4. c, d AlphaLISA assay to determine the formation of BCL-xL/PROTAC/VHL-ELOC-ELOB (VCB) (c) or BCL-2/PROTAC/VCB (d) ternary complex in a cell-free condition. Data represent the mean of a single experiment with 2 technical replicates. Similar results were obtained in one more independent experiment. e, f NanoBRET assays evaluating intracellular BCL-xL/PROTAC/VCB (e) or BCL-2/PROTAC/VCB (f) ternary complex formation. 293T cells were transiently transfected with HiBit-BCL-xL, LgBit, and HaloTag-VHL or HiBit-BCL-2, LgBit, and HaloTag-VHL and then treated with a serial dilution of the indicated compounds for 6 h. Data are expressed as mean ± s.e.m. of three biological replicates. Source data are provided as a Source data file.
Fig. 2 Synthesis and evaluation of a series of BCL-xL and/or BCL-2 degraders. a The structure of the BCL-xL and/or BCL-2 degraders (racemic mixture). b The BCL-xL and BCL-2 degradation profiling of the PROTACs evaluated in 293T cells after the cells were treated with four different concentrations of indicated PROTACs for 16 h. The DC50 and Dmax were calculated using Prism based on the ImageJ-quantified western blots in Supplementary Fig. 4. c, d AlphaLISA assay to determine the formation of BCL-xL/PROTAC/VHL-ELOC-ELOB (VCB) (c) or BCL-2/PROTAC/VCB (d) ternary complex in a cell-free condition. Data represent the mean of a single experiment with 2 technical replicates. Similar results were obtained in one more independent experiment. e, f NanoBRET assays evaluating intracellular BCL-xL/PROTAC/VCB (e) or BCL-2/PROTAC/VCB (f) ternary complex formation. 293T cells were transiently transfected with HiBit-BCL-xL, LgBit, and HaloTag-VHL or HiBit-BCL-2, LgBit, and HaloTag-VHL and then treated with a serial dilution of the indicated compounds for 6 h. Data are expressed as mean ± s.e.m. of three biological replicates. Source data are provided as a Source data file.CC BY · PubMed Central
Fig. 3 753b is a dual BCL-xL/2 degrader. a The structure of 753a and 753b. b The potency of 753a and 753b in degrading BCL-xL/2 was evaluated in 293T cells by immunoblots after the cells were treated with different concentrations of 753a or 753b for 16 h. c Comparison of the potencies of 753b and DT2216 in degrading BCL-xL/2 by immunoblots in 293T cells after the cells were treated with different concentrations of 753b or DT2216 for 16 h. d The time course of 753b-mediated BCL-xL/2 degradation was evaluated in 293T cells by immunoblots after the cells were treated with 1 μM 753b for various time points as indicated. Representative immunoblots are shown in (a–d) and β-actin was used as a loading control in all immunoblot analyses. The normalized protein content in the immunoblots is presented as mean values ± s.e.m. (n = 3 biologically independent experiments) in the bar graph (bottom panel). e, f The formation of BCL-xL/PROTAC/VCB (e) and BCL-2/PROTAC/VCB (f) ternary complexes in a cell-free condition was determined by AlphaLISA assay. Data represent the mean of a single experiment with 2 technical replicates. Similar results were obtained in one more independent experiment. g, h NanoBRET assays showed that 753b forms more stable ternary complexes with BCL-xL (g) and BCL-2 (h) in 293T cells compared with 753a. 293T cells were transiently transfected with HiBit-BCL-xL, LgBit, and HaloTag-VHL or HiBit-BCL-2, LgBit, and HaloTag-VHL and then treated with a serial dilution of 753a or 753b for 6 h. Data are expressed as mean of three biological replicates. i The different conformations of linker methyl groups on ABT263 observed in X-ray crystal structure of BCL-xL/ABT263 complex (PDB 4QNQ). Source data are provided as a Source data file.
Fig. 3 753b is a dual BCL-xL/2 degrader. a The structure of 753a and 753b. b The potency of 753a and 753b in degrading BCL-xL/2 was evaluated in 293T cells by immunoblots after the cells were treated with different concentrations of 753a or 753b for 16 h. c Comparison of the potencies of 753b and DT2216 in degrading BCL-xL/2 by immunoblots in 293T cells after the cells were treated with different concentrations of 753b or DT2216 for 16 h. d The time course of 753b-mediated BCL-xL/2 degradation was evaluated in 293T cells by immunoblots after the cells were treated with 1 μM 753b for various time points as indicated. Representative immunoblots are shown in (a–d) and β-actin was used as a loading control in all immunoblot analyses. The normalized protein content in the immunoblots is presented as mean values ± s.e.m. (n = 3 biologically independent experiments) in the bar graph (bottom panel). e, f The formation of BCL-xL/PROTAC/VCB (e) and BCL-2/PROTAC/VCB (f) ternary complexes in a cell-free condition was determined by AlphaLISA assay. Data represent the mean of a single experiment with 2 technical replicates. Similar results were obtained in one more independent experiment. g, h NanoBRET assays showed that 753b forms more stable ternary complexes with BCL-xL (g) and BCL-2 (h) in 293T cells compared with 753a. 293T cells were transiently transfected with HiBit-BCL-xL, LgBit, and HaloTag-VHL or HiBit-BCL-2, LgBit, and HaloTag-VHL and then treated with a serial dilution of 753a or 753b for 6 h. Data are expressed as mean of three biological replicates. i The different conformations of linker methyl groups on ABT263 observed in X-ray crystal structure of BCL-xL/ABT263 complex (PDB 4QNQ). Source data are provided as a Source data file.CC BY · PubMed Central
Fig. 4 753b degrades BCL-xL/2 in a VHL- and proteasome-dependent manner. a Pretreatment with ABT263 or VHL-L blocks the BCL-xL/2 degradation induced by 753b in 293T cells. b CRISPR knockout of VHL blocks 753b-induced BCL-xL/2 degradation as shown in wild type (WT) and VHL knockout (VHL-ko) 293T cells. c Inhibition of CRLVHLneddylation with MLN4924 blocks BCL-xL/2 degradation induced by 753b in 293T cells. d Inhibition of proteasomes with MG132, but not that of caspases with the pan-caspase inhibitor QVD, blocks 753b-induced BCL-xL/2 degradation in 293T cells. Source data are provided as a Source data file.
Fig. 4 753b degrades BCL-xL/2 in a VHL- and proteasome-dependent manner. a Pretreatment with ABT263 or VHL-L blocks the BCL-xL/2 degradation induced by 753b in 293T cells. b CRISPR knockout of VHL blocks 753b-induced BCL-xL/2 degradation as shown in wild type (WT) and VHL knockout (VHL-ko) 293T cells. c Inhibition of CRLVHLneddylation with MLN4924 blocks BCL-xL/2 degradation induced by 753b in 293T cells. d Inhibition of proteasomes with MG132, but not that of caspases with the pan-caspase inhibitor QVD, blocks 753b-induced BCL-xL/2 degradation in 293T cells. Source data are provided as a Source data file.CC BY · PubMed Central
Fig. 5 Identification of the key accessible surface lysines on BCL-xL and BCL-2. a 753b degrades WT BCL-xL and K20-only and K87-only BCL-xL mutants, but not K16-only BCL-xL mutant, in 293T cells. K16-only, K20-only, and K87-only represent that all the surface lysines on BCL-xL are mutated to arginines except K16, K20, and K87, respectively. Representative immunoblots of Flag-BCL-xL WT and mutants are shown in the top panel. Statistical significance was calculated with unpaired two-tailed Student’s t-tests comparing DMSO- to 753b-treated samples. *P < 0.05; **P < 0.01; P values were 0.0030, 0.0070, and 0.035 for WT, K20-only, and K87-only BCL-xL, respectively. b 753b degrades WT BCL-2 and K22R BCL-2 mutant, but not K17R and K17/22R BCL-2 mutants, in 293T cells. K17R and K22R represents that K17 and K22 on BCL-2 are respectively mutated to arginine; K17/22R means both K17 and K22 on BCL-2 are mutated to arginine. Representative immunoblots of Flag-BCL-2 WT and mutants are shown in the top panel. The quantification of the relative Flag-BCL-xL or Flag-BCL-2 protein content in the immunoblots is presented as mean values ± s.e.m. (n = 3 biologically independent experiments) in the bottom panel. Statistical significance was calculated with unpaired two-tailed Student’s t-tests comparing DMSO- to 753b-treated samples. **P < 0.01; P values were 0.0043 and 0.0020 for WT and K22R BCL-2 mutant, respectively. c 753b induces polyubiquitination of WT, K20-only, and K87-only BCL-xL, but not K16-only BCL-xL in 293 T cells. d 753b induces polyubiquitination of WT and K22R BCL-2, but not K17R and K17/22R BCL-2 in 293T cells. For c, d 293T cells were co-transfected as indicated with Flag-BCL-xL WT or mutants and HA-tagged ubiquitin (HA-Ub) plasmids (c) or Flag-BCL-2 WT or mutants and HA-Ub plasmids (d). After 36 h, cells were pre-treated with MG132 (10 µM) for 2 h and then treated with or without 753b (0.1 µM for BCL-xL and 1 uM for BCL-2) for 5 h. The Flag-tagged proteins were immunoprecipitated (IP) and immunoblotted with HA or Flag antibody. The input results are shown in Supplementary Fig. 9. Data are a representative of two independent experiments. Source data are provided as a Source data file.
Fig. 5 Identification of the key accessible surface lysines on BCL-xL and BCL-2. a 753b degrades WT BCL-xL and K20-only and K87-only BCL-xL mutants, but not K16-only BCL-xL mutant, in 293T cells. K16-only, K20-only, and K87-only represent that all the surface lysines on BCL-xL are mutated to arginines except K16, K20, and K87, respectively. Representative immunoblots of Flag-BCL-xL WT and mutants are shown in the top panel. Statistical significance was calculated with unpaired two-tailed Student’s t-tests comparing DMSO- to 753b-treated samples. *P < 0.05; **P < 0.01; P values were 0.0030, 0.0070, and 0.035 for WT, K20-only, and K87-only BCL-xL, respectively. b 753b degrades WT BCL-2 and K22R BCL-2 mutant, but not K17R and K17/22R BCL-2 mutants, in 293T cells. K17R and K22R represents that K17 and K22 on BCL-2 are respectively mutated to arginine; K17/22R means both K17 and K22 on BCL-2 are mutated to arginine. Representative immunoblots of Flag-BCL-2 WT and mutants are shown in the top panel. The quantification of the relative Flag-BCL-xL or Flag-BCL-2 protein content in the immunoblots is presented as mean values ± s.e.m. (n = 3 biologically independent experiments) in the bottom panel. Statistical significance was calculated with unpaired two-tailed Student’s t-tests comparing DMSO- to 753b-treated samples. **P < 0.01; P values were 0.0043 and 0.0020 for WT and K22R BCL-2 mutant, respectively. c 753b induces polyubiquitination of WT, K20-only, and K87-only BCL-xL, but not K16-only BCL-xL in 293 T cells. d 753b induces polyubiquitination of WT and K22R BCL-2, but not K17R and K17/22R BCL-2 in 293T cells. For c, d 293T cells were co-transfected as indicated with Flag-BCL-xL WT or mutants and HA-tagged ubiquitin (HA-Ub) plasmids (c) or Flag-BCL-2 WT or mutants and HA-Ub plasmids (d). After 36 h, cells were pre-treated with MG132 (10 µM) for 2 h and then treated with or without 753b (0.1 µM for BCL-xL and 1 uM for BCL-2) for 5 h. The Flag-tagged proteins were immunoprecipitated (IP) and immunoblotted with HA or Flag antibody. The input results are shown in Supplementary Fig. 9. Data are a representative of two independent experiments. Source data are provided as a Source data file.CC BY · PubMed Central
Fig. 6 Computational models of the CRLVHL/753b/BCL-xL/E2-Ub/RBX1 and CRLVHL/753b/BCL-2/E2-Ub/RBX1 complexes. a The superimposition of the modeled ternary complexes of VCB/753b/BCL-xL and VCB/753b/BCL-2. The Cα RMSD of BCL-xL and BCL-2 is 6.7 Å. b The computational model of catalytic transition state of CRLVHL/753b/BCL-xL/UbcH5B-Ub/RBX1 with UbcH5B contacting with K87 of BCL-xL. The K87 of BCL-xL, C85, and D117 of UbcH5B are colored in blue, orange, and red, respectively. c The computational model of catalytic transition state of CRLVHL/753b/BCL-xL/UbcH5B-Ub/RBX1 with UbcH5B contacting with K20 of BCL-xL. d The computational model of catalytic transition state of CRLVHL/753b/BCL-2/UbcH5B-Ub/RBX1 with UbcH5B contacting with K17 of BCL-2.
Fig. 6 Computational models of the CRLVHL/753b/BCL-xL/E2-Ub/RBX1 and CRLVHL/753b/BCL-2/E2-Ub/RBX1 complexes. a The superimposition of the modeled ternary complexes of VCB/753b/BCL-xL and VCB/753b/BCL-2. The Cα RMSD of BCL-xL and BCL-2 is 6.7 Å. b The computational model of catalytic transition state of CRLVHL/753b/BCL-xL/UbcH5B-Ub/RBX1 with UbcH5B contacting with K87 of BCL-xL. The K87 of BCL-xL, C85, and D117 of UbcH5B are colored in blue, orange, and red, respectively. c The computational model of catalytic transition state of CRLVHL/753b/BCL-xL/UbcH5B-Ub/RBX1 with UbcH5B contacting with K20 of BCL-xL. d The computational model of catalytic transition state of CRLVHL/753b/BCL-2/UbcH5B-Ub/RBX1 with UbcH5B contacting with K17 of BCL-2.CC BY · PubMed Central
Fig. 7 753b is a more potent anti-leukemic agent than DT2216. a Kasumi-1 AML cells are primarily dependent on BCL-xL/2 for survival. Viability of Kasumi-1 AML cells was measured after the cells were incubated with serially diluted ABT199 (a selective BCL-2 inhibitor), ABT263 (a dual BCL-xL/BCL-2 inhibitor), A1331872 (a selective BCL-xL inhibitor), or S63845 (a selective MCL-1 inhibitor), for 72 h. IC50 values for ABT199, ABT263, A1331872, and S63845 are 866.9, 156.5, 3587.0, and 527.4 nM, respectively. b Potencies of 753b and DT2216 in degrading BCL-xL/2 by immunoblots in Kasumi-1 cells after 16 h treatment. Tubulin was used as an equal loading control. The normalized protein content in the immunoblots is presented as mean values (n = 2 biologically independent experiments) in the bar graph (right panel). c–h 753b is more potent against Kasumi-1 cells than DT2216. c Viability of Kasumi-1 cells was measured after the cells were incubated with serially diluted DT2216, 753b, ABT199, or ABT263 for 72 h. IC50 values for DT2216, 753b, ABT199, and ABT263 are 1147.0, 59.64, 1130.0, and 130.9 nM, respectively. For the viability assays, the data are presented as mean ± s.d. from three replicate cell cultures in a representative experiment. Similar results were also observed in two additional independent experiments. d Representative immunoblot analyses of Kasumi-1 cells 16 h after treatment as indicated. Tubulin was used as an equal loading control. e Representative flow cytometry analyses of apoptosis using Annexin-V and DAPI staining in Kasumi-1 cells 24 h after treatment as indicated. f Representative flow cytometry analyses of apoptosis using Annexin-V and DAPI staining in Kasumi-1 cells after incubated with or without the pan-caspase inhibitor QVD (10 μM) for 2 h prior to being treated with DMSO (Veh) or 0.1 μM DT2216, 753b, ABT199, or ABT263 for 24 h. g, h Percentages of apoptotic cells in Kasium-1 cells analyzed by flow cytometry as shown in (e, f) are presented as mean ± s.d. from three replicate cell cultures in a representative experiment. Data are representative of two independent experiments. The dotted line represents the value from DMSO-treated cells. Source data are provided as a Source data file.
Fig. 7 753b is a more potent anti-leukemic agent than DT2216. a Kasumi-1 AML cells are primarily dependent on BCL-xL/2 for survival. Viability of Kasumi-1 AML cells was measured after the cells were incubated with serially diluted ABT199 (a selective BCL-2 inhibitor), ABT263 (a dual BCL-xL/BCL-2 inhibitor), A1331872 (a selective BCL-xL inhibitor), or S63845 (a selective MCL-1 inhibitor), for 72 h. IC50 values for ABT199, ABT263, A1331872, and S63845 are 866.9, 156.5, 3587.0, and 527.4 nM, respectively. b Potencies of 753b and DT2216 in degrading BCL-xL/2 by immunoblots in Kasumi-1 cells after 16 h treatment. Tubulin was used as an equal loading control. The normalized protein content in the immunoblots is presented as mean values (n = 2 biologically independent experiments) in the bar graph (right panel). c–h 753b is more potent against Kasumi-1 cells than DT2216. c Viability of Kasumi-1 cells was measured after the cells were incubated with serially diluted DT2216, 753b, ABT199, or ABT263 for 72 h. IC50 values for DT2216, 753b, ABT199, and ABT263 are 1147.0, 59.64, 1130.0, and 130.9 nM, respectively. For the viability assays, the data are presented as mean ± s.d. from three replicate cell cultures in a representative experiment. Similar results were also observed in two additional independent experiments. d Representative immunoblot analyses of Kasumi-1 cells 16 h after treatment as indicated. Tubulin was used as an equal loading control. e Representative flow cytometry analyses of apoptosis using Annexin-V and DAPI staining in Kasumi-1 cells 24 h after treatment as indicated. f Representative flow cytometry analyses of apoptosis using Annexin-V and DAPI staining in Kasumi-1 cells after incubated with or without the pan-caspase inhibitor QVD (10 μM) for 2 h prior to being treated with DMSO (Veh) or 0.1 μM DT2216, 753b, ABT199, or ABT263 for 24 h. g, h Percentages of apoptotic cells in Kasium-1 cells analyzed by flow cytometry as shown in (e, f) are presented as mean ± s.d. from three replicate cell cultures in a representative experiment. Data are representative of two independent experiments. The dotted line represents the value from DMSO-treated cells. Source data are provided as a Source data file.CC BY · PubMed Central

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  1. Ester, M., Kriegel, H.-P., Sander, J. & Xu, X. in Kdd, Vol. 96, 226-231 (1996).

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Authors
Lv, D., Pal, P., Liu, X., Jia, Y., Thummuri, D., Zhang, P., Hu, W., Pei, J., Zhang, Q., Zhou, S., Khan, S., Zhang, X., Hua, N., Yang, Q., Arango, S., Zhang, W., Nayak, D., Olsen, S. K., Weintraub, S. T., Hromas, R., Konopleva, M., Yuan, Y., Zheng, G., & Zhou, D.
Affiliations
  • University of Florida (US)
  • The University of Texas MD Anderson Cancer Center (US)
  • The University of Texas at San Antonio Health Science Center (US)
Corresponding author
  • Yuan, Yaxia
  • Zheng, Guangrong
  • Zhou, Daohong
ORCID
Venue
Nature Communications
Publisher
Springer Science and Business Media LLC
Published
2021-11-25
Volume
12
Issue
1
Pages
6896-6896
ISSN
2041-1723 (ISSN-L)
DOI
10.1038/s41467-021-27210-x
PMC
PMC8617031
OpenAlex
W3215130076
Cited by
158
References
55
Citations by year
2026: 27 · 2025: 38 · 2024: 41 · 2023: 22 · 2022: 30
FWCI
7.30
Type
journal article
Language
en
Open access
Licence
Locations
  • Nature Communications (publishedVersion, cc-by)
  • PubMed Central (publishedVersion, cc-by)
  • PubMed (publishedVersion)
  • RePEc: Research Papers in Economics (submittedVersion)
  • DOAJ (DOAJ: Directory of Open Access Journals) (submittedVersion, cc-by-sa)
Funders
  • U.S. Department of Health & Human Services | National Institutes of Health (R01 CA241191, R01 AG063801, R01 CA205224, R01 GM115568, R01 GM128731, R01 CA200673, R01 CA203834)
  • Cancer Prevention and Research Institute of Texas (RR200030)
Topics
  • Life Sciences › Biochemistry, Genetics and Molecular Biology › Molecular Biology › Protein Degradation and Inhibitors
  • Life Sciences › Biochemistry, Genetics and Molecular Biology › Molecular Biology › Ubiquitin and proteasome pathways
  • Life Sciences › Biochemistry, Genetics and Molecular Biology › Molecular Biology › Cell death mechanisms and regulation
Keywords
PROTACs, Bcl-xL, Bcl-2, UbcH5b, RBX1, ubiquitination, navitoclax, leukemia, protein degradation
MeSH terms
Antineoplastic Agents, Cell Line, Cell Survival, Humans, Leukemia, Lysine, Models, Molecular, Protein Binding, Protein Conformation, Proto-Oncogene Proteins c-bcl-2, Ubiquitin-Conjugating Enzymes, Ubiquitin-Protein Ligases, Proteasome Endopeptidase Complex, bcl-X Protein, Von Hippel-Lindau Tumor Suppressor Protein, Small Molecule Libraries, Ubiquitination, Proteolysis

About these checks

Registry, retraction, Cites and statistics results are computed, not judged, with one exception: a printed reference that no index lists is read once by a model before it counts as a citation error. Claims are extracted by a model and kept only when their quote appears verbatim in the text; each cited source is then checked, with a model’s help, for whether it says what the claim relies on. Those verdicts can be wrong — read the claim’s quote and the cited source before relying on one.

Statistics and arithmetic are recomputed, not judged.

Confidence bands are shown only for fields calibrated on our annotated test set. None is calibrated yet, so no band is shown.

What counts as a claim ›How we measure confidence ›How we check this ›

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