Keap1 Recruits Neh2 through Binding to ETGE and DLG Motifs: Characterization of the Two-Site Molecular Recognition Model
Open accessAbstract
The expression of the phase 2 detoxification enzymes and antioxidant proteins is induced at the transcriptional level by Nrf2 and negatively regulated at the posttranslational level by Keap1 through protein-protein interactions with and subsequent proteolysis of Nrf2. We found that the Neh2 domain of Nrf2 is an intrinsically disordered but biologically active regulatory domain containing a 33-residue central alpha-helix followed by a mini antiparallel beta-sheet. Isothermal calorimetry analysis indicated that one Neh2 molecule interacts with two molecules of Keap1 via two binding sites, the stronger binding ETGE motif and the weaker binding DLG motif. Nuclear magnetic resonance titration study showed that these two motifs of the Neh2 domain bind to an overlapping site on the bottom surface of the beta-propeller structure of Keap1. In contrast, the central alpha-helix of the Neh2 domain does not have any observable affinity to Keap1, suggesting that this region may serve as a bridge connecting the two motifs for the association with the two beta-propeller structures of a dimer of Keap1. Based on these observations, we propose that Keap1 recruits Nrf2 by the ETGE motif and that the DLG motif of the Neh2 domain locks its lysine-rich central alpha-helix in a correct position to benefit ubiquitin signaling.
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Keap1 recruits Nrf2 first through binding of one Keap1 molecule to the high-affinity ETGE motif, and subsequent binding of the other Keap1 molecule at the low-affinity DLG motif locks Nrf2 in place for ubiquitination by Cul3 and degradation by the 26S proteasome.
“Keap1 recruits Nrf2 first through the binding of one Keap1 molecule to the high-affinity ETGE motif within the Nrf2’s Neh2 domain. Subsequent binding of the other Keap1 molecule at the low-affinity DLG motif locks Nrf2 in place by orienting the lysine residues within Neh2 in the correct position for ubiquitination by Cul3 and degradation by the 26S proteasome [58,65].”
From Nrf2 and Oxidative Stress: A General Overview of Mechanisms and Implications in Human Disease (2022) · cites this paper as [58]
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Self-citationActivation of Nrf2-antioxidant signaling attenuates NFκB-inflammatory response and elicits apoptosis
Li, W., Khor, T. O., Xu, C., Shen, G., Jeong, W.-S., Yu, S., & Kong, A.-N. (2008) · Biochemical Pharmacology · doi:10.1016/j.bcp.2008.07.017 · Cited by 738 · see citing works
“, a high affinity phase I binding, mediated by the ETGE motif and a low affinity phase II binding mediated by the DLG motif, has been distinguished [8].”
BackgroundShibata, T., Ohta, T., Tong, K. I., Kokubu, A., Odogawa, R., Tsuta, K., Asamura, H., Yamamoto, M., & Hirohashi, S. (2008) · Proceedings of the National Academy of Sciences · doi:10.1073/pnas.0806268105 · Cited by 728 · see citing works
“The double glycine repeat and the C-terminal region of Keap1 (hereafter refer as to Keap1-DC) recognize both the DLG and ETGE motifs of Nrf2 (20, 21).”
BackgroundSelf-citationDirect Interaction between Nrf2 and p21Cip1/WAF1 Upregulates the Nrf2-Mediated Antioxidant Response
Chen, W., Sun, Z., Wang, X.-J., Jiang, T., Huang, Z., Fang, D., & Zhang, D. D. (2009) · Molecular Cell · doi:10.1016/j.molcel.2009.04.029 · Cited by 638 · see citing works
“It is believed that the Keap1 homodimer binds both the 29 DLG and 79 ETGE motifs in Nrf2 to align the seven ubiquitin-accepting lysine residues between these motifs into a conformation suitable for ubiquitin conjugation (McMahon et al., 2006; Tong et al., 2006, 2007; Zhang et al., 2004).”
BackgroundInfluentialLAU, A., VILLENEUVE, N., Sun, Z., Wong, P. K., & Zhang, D. (2008) · Pharmacological Research · doi:10.1016/j.phrs.2008.09.003 · Cited by 645 · see citing works
“hat bind the Kelch domain of Keap1, which is the proposed hinge nd latch two-site binding model [54,55].”
Backgroundp62 links autophagy and Nrf2 signaling
Jiang, T., Harder, B. G., de la Vega, M. R., Wong, P. K., Chapman, E., & Zhang, D. D. (2015) · Free Radical Biology and Medicine · doi:10.1016/j.freeradbiomed.2015.06.014 · Cited by 642 · see citing works
“According to the hinge and latch model, the ETGE motif has a higher affinity for Keap1 than the DLG motif, which causes the latter to dynamically associate and dissociate from Keap1 to generate oscillating “closed” (associated) and “open” (dissociated) conformations [29–33].”
Emerging roles of Nrf2 and phase II antioxidant enzymes in neuroprotection
Zhang, M., An, C., Gao, Y., Leak, R. K., Chen, J., & Zhang, F. (2012) · Progress in Neurobiology · doi:10.1016/j.pneurobio.2012.09.003 · Cited by 622 · see citing works
“The ETGF motif (KD ≈ 1000 nmol/L) has a higher affinity for Keap1 than the DLG motif (KD ≈ 5.3 nmol/L) (Tong et al., 2006).”
BackgroundInfluentialActivators and Inhibitors of NRF2: A Review of Their Potential for Clinical Development
Robledinos-Antón, N., Fernández-Ginés, R., Manda, G., & Cuadrado, A. (2019) · Oxidative Medicine and Cellular Longevity · doi:10.1155/2019/9372182 · Cited by 608 · see citing works
Structural basis of Keap1 interactions with Nrf2
Canning, P., Sorrell, F. J., & Bullock, A. N. (2015) · Free Radical Biology and Medicine · doi:10.1016/j.freeradbiomed.2015.05.034 · Cited by 605 · see citing works
Structure of the Keap1:Nrf2 interface provides mechanistic insight into Nrf2 signaling
Lo, S., Li, X., Henzl, M. T., Beamer, L. J., & Hannink, M. (2006) · The EMBO Journal · doi:10.1038/sj.emboj.7601243 · Cited by 604 · see citing works
“…Biology Organization The EMBO Journal VOL 25 | NO 15 | 2006 3613 second Keap1 binding site in Nrf2, containing the consensus sequence LxxQDxDLG and located at a distance of approximately 50 amino acids on the N-terminal side of the DxETGE motif, has recently been identified (Tong et al, 2006).”
BackgroundMethodInfluentialBasic Principles and Emerging Concepts in the Redox Control of Transcription Factors
Brigelius-Flohé, R., & Flohé, L. (2011) · Antioxidants and Redox Signaling · doi:10.1089/ars.2010.3534 · Cited by 577 · see citing works
“The Neh2 domain is locked in a position suitable for ubiquitination of the Lys residues (325, 485) which, hence, act as ubiquitin acceptors (532).”
BackgroundNrf2 signaling: An adaptive response pathway for protection against environmental toxic insults
Osburn, W. O., & Kensler, T. W. (2007) · Mutation Research/Reviews in Mutation Research · doi:10.1016/j.mrrev.2007.11.006 · Cited by 549 · see citing works
“Normally, under basal conditions Nrf2 is bound to Keap1 in the cytoplasm due to an interaction between a single Nrf2 protein and a Keap1 dimer[3].”
BackgroundCancer Chemoprevention Mechanisms Mediated Through the Keap1–Nrf2 Pathway
Hayes, J. D., McMahon, M. J., Chowdhry, S., & Dinkova‐Kostova, A. T. (2010) · Antioxidants and Redox Signaling · doi:10.1089/ars.2010.3221 · Cited by 540 · see citing works
The KEAP1–NRF2 System in Cancer
Taguchi, K., & Yamamoto, M. (2017) · Frontiers in Oncology · doi:10.3389/fonc.2017.00085 · Cited by 527 · see citing works
“Proc Natl Acad Sci U S A (2011) 108(4):1433–8.”
BackgroundInfluentialSelf-citationPhysiological Significance of Reactive Cysteine Residues of Keap1 in Determining Nrf2 Activity
Yamamoto, T., Suzuki, T., Kobayashi, A., Wakabayashi, J., Maher, J. M., Motohashi, H., & Yamamoto, M. P. (2008) · Molecular and Cellular Biology · doi:10.1128/mcb.01704-07 · Cited by 515 · see citing works
“Although it was shown previously that the BTB domain mediates Keap1 dimerization (40) and that Keap1 remains as a homodimer in a solution (20, 32), the physiological contribution of the BTB domain to the function of Keap1 was still poorly understood.”
BackgroundInfluentialSelf-citationNrf2/Keap1/ARE signaling: Towards specific regulation
Ulasov, A. V., Rosenkranz, A. A., Georgiev, G. P., & Sobolev, A. S. (2021) · Life Sciences · doi:10.1016/j.lfs.2021.120111 · Cited by 520 · see citing works
“The Keap1 Kelch domain constitutes a sixbladed β-propeller structure, while the Neh2 domain is intrinsically disordered [206].”
BackgroundDiscovery of the Negative Regulator of Nrf2, Keap1: A Historical Overview
Itoh, K., Mimura, J., & Yamamoto, M. P. (2010) · Antioxidants and Redox Signaling · doi:10.1089/ars.2010.3222 · Cited by 513 · see citing works
“While the N-terminal subdomain (residues 16–31) contains hydrophobic amino acid residues that can potentially form an amphipathic a -helix (47, 106), the central portion of the domain is rich in hydrophilic residues that may also adopt an a -helical structure (106) (Fig.”
Self-citationReactive Oxygen Species (ROS) Regulates Different Types of Cell Death by Acting as a Rheostat
Villalpando-Rodriguez, G. E., & Gibson, S. B. (2021) · Oxidative Medicine and Cellular Longevity · doi:10.1155/2021/9912436 · Cited by 499 · see citing works
“Conversely, ROS-mediated signaling could activate procell death pathways such a JNK signaling [13, 43, 53].”
BackgroundMcMahon, M. J., Thomas, N., Itoh, K., Yamamoto, M. P., & Hayes, J. D. (2006) · Journal of Biological Chemistry · doi:10.1074/jbc.m601119200 · Cited by 489 · see citing works
Self-citationOrchestrating Redox Signaling Networks through Regulatory Cysteine Switches
Paulsen, C. E., & Carroll, K. S. (2009) · ACS Chemical Biology · doi:10.1021/cb900258z · Cited by 478 · see citing works
Nrf2 at the heart of oxidative stress and cardiac protection
Chen, Q. M., & Maltagliati, A. J. (2017) · Physiological Genomics · doi:10.1152/physiolgenomics.00041.2017 · Cited by 474 · see citing works
“Binding to both ETGE and DLG domains of Nrf2 is essential for Keap1-mediated ubiquitination, as such motion induces a conformation change of Nrf2 protein within the E3 ubiquitin ligase complex that allows Nrf2 to be polyubiquitinated (150, 215, 217).”
BackgroundNrf2 in cancers: A double‐edged sword
Wu, S., Lu, H., & Bai, Y. (2019) · Cancer Medicine · doi:10.1002/cam4.2101 · Cited by 473 · see citing works
Two-site substrate recognition model for the Keap1-Nrf2 system: A hinge and latch mechanism
Tong, K. I., Kobayashi, A., Katsuoka, F., & Yamamoto, M. (2006) · Biological Chemistry · doi:10.1515/bc.2006.164 · Cited by 473 · see citing works
“NMR structural analysis revealed that Neh2 is composed of a 33-aa central a-helix, followed by a small anti-parallel b-sheet (Tong et al., 2006).”
BackgroundInfluentialSelf-citationThe complexity of the Nrf2 pathway: beyond the antioxidant response
Huang, Y. S., Li, W., Su, Z., & Kong, A. T. (2015) · The Journal of Nutritional Biochemistry · doi:10.1016/j.jnutbio.2015.08.001 · Cited by 449 · see citing works
“Nrf2 has two binding motifs in the Neh2 domain, the ETGE and DLG motifs, and recruits two molecules of Keap1 in the absence of stimuli [47, 48].”
BackgroundTong, K. I., Padmanabhan, B., Kobayashi, A., Shang, C., Hirotsu, Y., Yokoyama, S., & Yamamoto, M. P. (2007) · Molecular and Cellular Biology · doi:10.1128/mcb.00753-07 · Cited by 447 · see citing works
“Our previous nuclear magnetic resonance chemical shift perturbation study suggested that both DLG and ETGE interact with an overlapping binding surface on Keap1-DC composed of conserved arginine residues (39).”
BackgroundMethodInfluentialSelf-citationSignal amplification in the KEAP1-NRF2-ARE antioxidant response pathway
Liu, S., Pi, J., & Zhang, Q. (2022) · Redox Biology · doi:10.1016/j.redox.2022.102389 · Cited by 445 · see citing works
Villeneuve, N. F., Lau, A., & Zhang, D. D. (2010) · Antioxidants and Redox Signaling · doi:10.1089/ars.2010.3211 · Cited by 423 · see citing works
Suzuki, T., & Yamamoto, M. P. (2017) · Journal of Biological Chemistry · doi:10.1074/jbc.r117.800169 · Cited by 402 · see citing works
“This two-site recognition/binding mechanism of the Nrf2 Neh2 domain and Keap1 homodimer is essential for the efficient ubiquitination of Nrf2 (31, 33, 34).”
BackgroundMethodSelf-citationThe Keap1–Nrf2 system in cancers: stress response and anabolic metabolism
Mitsuishi, Y., Motohashi, H., & Yamamoto, M. P. (2012) · Frontiers in Oncology · doi:10.3389/fonc.2012.00200 · Cited by 394 · see citing works
“Two DC domains of the Keap1 homodimer associate with one molecule of Nrf2 (Tong et al., 2006).”
BackgroundSelf-citationCanonical and non-canonical mechanisms of Nrf2 activation
Silva‐Islas, C. A., & Maldonado, P. D. (2018) · Pharmacological Research · doi:10.1016/j.phrs.2018.06.013 · Cited by 393 · see citing works
Activation of NRF2 by Nitrosative Agents and H2O2 Involves KEAP1 Disulfide Formation
Fourquet, S., Guérois, R., Biard, D. S. F., & Tolédano, M. B. (2010) · Journal of Biological Chemistry · doi:10.1074/jbc.m109.051714 · Cited by 378 · see citing works
Proteasomal Degradation Is Transcriptionally Controlled by TCF11 via an ERAD-Dependent Feedback Loop
Steffen, J., Seeger, M., Koch, A., & Krüger, E. (2010) · Molecular Cell · doi:10.1016/j.molcel.2010.09.012 · Cited by 370 · see citing works
Keap1–Nrf2 Signaling: A Target for Cancer Prevention by Sulforaphane
Kensler, T. W., Egner, P. A., Agyeman, A. S., Visvanathan, K., Groopman, J. D., Chen, J., Chen, T.-Y., Fahey, J. W., & Talalay, P. (2012) · Topics in current chemistry · doi:10.1007/128_2012_339 · Cited by 363 · see citing works
Oncogenic NRF2 mutations in squamous cell carcinomas of oesophagus and skin
Kim, Y. R., Oh, J. E., Kim, M. S., Kang, M. R., Park, S. W., Han, J. Y., Eom, H., Yoo, N. J., & Lee, S. H. (2009) · The Journal of Pathology · doi:10.1002/path.2653 · Cited by 357 · see citing works
“KEAP1 normally exists as a homodimer, and one NRF molecule interacts with two KEAP1 molecules via two binding sites, the stronger binding the ETGE motif and the weaker binding the DLG motif [16,17].”
BackgroundNatural product-derived pharmacological modulators of Nrf2/ARE pathway for chronic diseases
Kumar, H., Kim, I., More, S. V., Kim, B.-W., & Choi, D. (2013) · Natural Product Reports · doi:10.1039/c3np70065h · Cited by 334 · see citing works
Keap1, the cysteine-based mammalian intracellular sensor for electrophiles and oxidants
Dinkova‐Kostova, A. T., Kostov, R. V., & Canning, P. (2016) · Archives of Biochemistry and Biophysics · doi:10.1016/j.abb.2016.08.005 · Cited by 332 · see citing works
“Nrf2 protein, known as the ETGE and DLG motifs [37].”
BackgroundSun, Z., Zhang, S., Chan, J. Y., & Zhang, D. D. (2007) · Molecular and Cellular Biology · doi:10.1128/mcb.00630-07 · Cited by 332 · see citing works
“In accordance with our data, Zhang and colleagues have recently demonstrated that Keap1 does not regulate Nrf1, although the Keap1 binding motif is conserved in Nrf1(49).”
BackgroundInfluentialThe Nrf2–Keap1 defence pathway: Role in protection against drug-induced toxicity
Copple, I. M., Goldring, C. E., Kitteringham, N. R., & Park, B. K. (2007) · Toxicology · doi:10.1016/j.tox.2007.10.029 · Cited by 328 · see citing works
Nrf2—a therapeutic target for the treatment of neurodegenerative diseases
Johnson, D. A., & Johnson, J. A. (2015) · Free Radical Biology and Medicine · doi:10.1016/j.freeradbiomed.2015.07.147 · Cited by 328 · see citing works
“When Keap1 senses oxidative or electrophilic stress, the low-affinity domain binding Nrf2 is abolished and proteosomal degradation of Nrf2 is disrupted [49].”
BackgroundAbed, D. A., Goldstein, M., Albanyan, H., Jin, H., & Hu, L. (2015) · Acta Pharmaceutica Sinica B · doi:10.1016/j.apsb.2015.05.008 · Cited by 330 · see citing works
Molecular Basis of Electrophilic and Oxidative Defense: Promises and Perils of Nrf2
Ma, Q., & He, X. (2012) · Pharmacological Reviews · doi:10.1124/pr.110.004333 · Cited by 317 · see citing works
“A series of peptide inhibitors were synthesized based on the Nrf2 Neh2 domain and the peptide sequences of p62 and prothymosin-α , which are known to bind Keap1 DC competitively with Nrf2 Neh2.”
InfluentialPGAM5 tethers a ternary complex containing Keap1 and Nrf2 to mitochondria
Lo, S.-C., & Hannink, M. (2008) · Experimental Cell Research · doi:10.1016/j.yexcr.2008.02.014 · Cited by 300 · see citing works
“Biophysical studies have suggested the existence of a second motif in Nrf2 that also binds to the Kelch domain of Keap1, leading to a model in which the Keap1 dimer binds a single molecule of Nrf2, resulting in a Keap1–Nrf2 complex with a 2:1 stoichiometry [58–60].”
BackgroundRedox Control of Microglial Function: Molecular Mechanisms and Functional Significance
Rojo, A. I., McBean, G. J., Cindrić, M., Egea, J., López, M. G., Rada, P., Žarković, N., & Cuadrado, A. (2014) · Antioxidants and Redox Signaling · doi:10.1089/ars.2013.5745 · Cited by 313 · see citing works
“It is thought that a single NRF2 polypeptide first binds one KEAP1 subunit through its high-affinity ETGE motif, and that this, in turn, enables the second low-affinity DLG motif to engage with the other Kelch-repeat domain of the remaining KEAP1 subunit (186, 313).”
Davies, T. G., Wixted, W. E., Coyle, J. E., Griffiths-Jones, C., Hearn, K., McMenamin, R., Norton, D., Rich, S. J., Richardson, C., Saxty, G., et al. (2016) · Journal of Medicinal Chemistry · doi:10.1021/acs.jmedchem.6b00228 · Cited by 322 · see citing works
Molecular Basis of the KEAP1-NRF2 Signaling Pathway
Suzuki, T., Takahashi, J. A., & Yamamoto, M. P. (2023) · Molecules and Cells · doi:10.14348/molcells.2023.0028 · Cited by 309 · see citing works
“The stoichiometry of KEAP1 and NRF2 within the KEAP1-NRF2 complex is 2:1, which is shown by means of isothermal calorimetry analysis (Tong et al., 2006a), nuclear magnetic resonance (NMR) titration analyses (Horie et al., 2021; Tong et al., 2006a), and an analytical centrifugation analysis (Iso et…”
BackgroundMethodInfluentialSelf-citationStructural basis for Cul3 protein assembly with the BTB-Kelch family of E3 ubiquitin ligases
Canning, P., Cooper, C. D. O., Krojer, T., Murray, J. W., Pike, A. C. W., Chaikuad, A., Keates, T., Thangaratnarajah, C., Hojzan, V., Marsden, B. D., et al. (2013) · Journal of Biological Chemistry · doi:10.1074/jbc.m112.437996 · Cited by 284 · see citing works
The KEAP1-NRF2 pathway: Targets for therapy and role in cancer
Adinolfi, S., Patinen, T., Deen, A. J., Pitkänen, S., Härkönen, J., Kansanen, E., Küblbeck, J., & Levonen, A. (2023) · Redox Biology · doi:10.1016/j.redox.2023.102726 · Cited by 301 · see citing works
“The interaction between NRF2 and KEAP1 is mediated via ETGE and DLG motifs at the Neh2 domain of NRF2 [14] (Fig.”
BackgroundThe Nrf2/HO‐1 Axis in Cancer Cell Growth and Chemoresistance
Furfaro, A. L., Traverso, N., Domenicotti, C., Piras, S., Moretta, L., Marinari, U. M., Pronzato, M. A., & Nitti, M. (2015) · Oxidative Medicine and Cellular Longevity · doi:10.1155/2016/1958174 · Cited by 295 · see citing works
Singh, S., Vrishni, S., Singh, B. K., Rahman, I., & Kakkar, P. (2010) · Free Radical Research · doi:10.3109/10715762.2010.507670 · Cited by 294 · see citing works
“Tong KI, Katoh Y, Kusunoki H, Itoh K, Tanaka T, Yamamoto [35]”
BackgroundThe Role of Nrf2 in Cardiovascular Function and Disease
Satta, S., Mahmoud, A. M., Wilkinson, F. L., Alexander, M. Y., & White, S. J. (2017) · Oxidative Medicine and Cellular Longevity · doi:10.1155/2017/9237263 · Cited by 299 · see citing works
“The Neh2 domain contains the Keap1 binding site (DLG and ETGE motifs), necessary for its cytoplasmic retention and degradation [55].”
BackgroundKEAP1 and done? Targeting the NRF2 pathway with sulforaphane
Dinkova‐Kostova, A. T., Fahey, J. W., Kostov, R. V., & Kensler, T. W. (2017) · Trends in Food Science & Technology · doi:10.1016/j.tifs.2017.02.002 · Cited by 286 · see citing works
“As reviewed elsewhere, dozens of targets and pathways have been identi fi ed as potential mediators of the chemoprotective actions of sulforaphane (Brown & Hampton, 2011; Hayes et al., 2008; Zhang, 2012): few have undergone serious validation.”
BackgroundInfluentialPark, C. H., Lee, S. L., Okamoto, T., Tanaka, T., & Yokozawa, T. (2012) · Evidence-based Complementary and Alternative Medicine · doi:10.1155/2012/587902 · Cited by 282 · see citing works
“On the other hand, oxidative stress induces alterations in the Nrf2 complex, and its gene transcription, such as that of HO-1, is enhanced [25].”
BackgroundSelf-citationNRF2 addiction in cancer cells
Kitamura, H., & Motohashi, H. (2018) · Cancer Science · doi:10.1111/cas.13537 · Cited by 275 · see citing works
Protein S-guanylation by the biological signal 8-nitroguanosine 3′,5′-cyclic monophosphate
Sawa, T., Zaki, M. H., Okamoto, T., Akuta, T., Tokutomi, Y., Kim‐Mitsuyama, S., Ihara, H., Kobayashi, A., Yamamoto, M. P., Fujii, S., et al. (2007) · Nature Chemical Biology · doi:10.1038/nchembio.2007.33 · Cited by 273 · see citing works
“His-tagged mouse recombinant Keap1 was expressed in Escherichia coli and purified as described elsewher”
MethodSelf-citationChen, Y., Yang, Z., Meng, M., Zhao, Y., Dong, N., Yan, H., Liu, L., Ding, M., Peng, H. B., & Shao, F. (2009) · Molecular Cell · doi:10.1016/j.molcel.2009.09.004 · Cited by 270 · see citing works
“In the best-studied Cul3/Keap1/Nrf2 complex, a Keap1 homodimer recognizes both the DLG and ETGE motif within one Nrf2 molecule (Tong et al., 2006).”
BackgroundShaping the Nrf2-ARE-related pathways in Alzheimer’s and Parkinson’s diseases
Fão, L., Mota, S. I., & Rego, A. C. (2019) · Ageing Research Reviews · doi:10.1016/j.arr.2019.100942 · Cited by 270 · see citing works
Role of Nrf2 and Its Activators in Respiratory Diseases
Liu, Q., Gao, Y., & Ci, X. (2019) · Oxidative Medicine and Cellular Longevity · doi:10.1155/2019/7090534 · Cited by 273 · see citing works
“Several mechanisms for the activation of Nrf2 by Keap1 have been proposed as the following up to now [17, 19, 20].”
BackgroundCrisman, E., Duarte, P., Daudén, E., Cuadrado, A., Rodrı́guez-Franco, M. I., López, M. G., & Leon, R. M. (2022) · Medicinal Research Reviews · doi:10.1002/med.21925 · Cited by 273 · see citing works
“There are only KEAP1 binding data and crystallographic information, but there is a lack of competition experiments with NRF2.”
BackgroundInfluentialBeyond repression of Nrf2: An update on Keap1
Kopacz, A., Klóska, D., Forman, H. J., Józkowicz, A., & Grochot‐Przeczek, A. (2020) · Free Radical Biology and Medicine · doi:10.1016/j.freeradbiomed.2020.03.023 · Cited by 262 · see citing works
“Many methods were implemented to study this interaction and to establish the minimal interacting fragment, including surface plasmon resonance [105], fluorescence polarisation assay [106], FRET [107], and ITC [108].”
MethodFukutomi, T., Takagi, K., Mizushima, T., Ohuchi, N., & Yamamoto, M. P. (2013) · Molecular and Cellular Biology · doi:10.1128/mcb.01191-13 · Cited by 265 · see citing works
“(B) A GST pulldown assay with 6 His-GST-tagged WT Neh2(1-56) or mutant forms thereof with purified recombinant Keap1-His.”
BackgroundMethodInfluentialSelf-citationStructure of the BTB Domain of Keap1 and Its Interaction with the Triterpenoid Antagonist CDDO
Cleasby, A., Yon, J. R., Day, P. J., Richardson, C. J., Tickle, I. J., Williams, P. A., Callahan, J. F., Carr, R. A. E., Concha, N. O., Kerns, J. K., et al. (2014) · PLoS ONE · doi:10.1371/journal.pone.0098896 · Cited by 263 · see citing works
Cullin-RING Ubiquitin Ligase Regulatory Circuits: A Quarter Century Beyond the F-Box Hypothesis
Harper, J. W., & Schulman, B. A. (2021) · Annual Review of Biochemistry · doi:10.1146/annurev-biochem-090120-013613 · Cited by 252 · see citing works
Hast, B. E., Goldfarb, D., Mulvaney, K. M., Hast, M. A., Siesser, P. F., Yan, F., Hayes, D. N., & Major, M. B. (2013) · Cancer Research · doi:10.1158/0008-5472.can-12-4400 · Cited by 259 · see citing works
“Crystallographic modeling revealed that NRF2 binds KEAP1 near the central pore of the KELCHβ-propeller (Figure 4E) (10, 13, 32).”
NRF2, a Superstar of Ferroptosis
Yan, R., Lin, B., Jin, W., Tang, L., Hu, S., & Cai, R. (2023) · Antioxidants · doi:10.3390/antiox12091739 · Cited by 260 · see citing works
“As mentioned above, under normal circumstances, two molecules of Keap1 bind to the ETGE and DLG motifs on the Neh2 domain of NRF2 via its Kelch repeat domain [44,59].”
Giudice, A., Arra, C., & Turco, M. C. (2010) · Methods in molecular biology · doi:10.1007/978-1-60761-738-9_3 · Cited by 254 · see citing works
“With regards to the importance of Nrf2 phosphorylation, two Nrf2 phosphorylation events have been recognized to modulate Nrf2 nuclear import/export in response to various chemopreventive agents (51).”
BackgroundInfluentialKeap1 facilitates p62-mediated ubiquitin aggregate clearance via autophagy
Fan, W., Tang, Z., Chen, D., Moughon, D., Ding, X., Chen, S., Zhu, M., & Zhong, Q. (2010) · Autophagy · doi:10.4161/auto.6.5.12189 · Cited by 251 · see citing works
Marcotte, D. J., Zeng, W., Hus, J., McKenzie, A., Hession, C., Jin, P., Bergeron, C., Lugovskoy, A. A., Enyedy, I. J., Cuervo, H., et al. (2013) · Bioorganic & Medicinal Chemistry · doi:10.1016/j.bmc.2013.04.019 · Cited by 245 · see citing works
Galea, C. A., Wang, Y., Sivakolundu, S. G., & Kriwacki, R. W. (2008) · Biochemistry · doi:10.1021/bi8006803 · Cited by 243 · see citing works
Rojo, A. I., de Sagarra, M. R., & Cuadrado, A. (2007) · Journal of Neurochemistry · doi:10.1111/j.1471-4159.2007.05124.x · Cited by 241 · see citing works
Hammad, M., Raftari, M., Cesário, R., Salma, R., Godoy, P. R. D. V., Emami, S. N., & Haghdoost, S. (2023) · Antioxidants · doi:10.3390/antiox12071371 · Cited by 245 · see citing works
“One of the most-known hyper-reactive cysteines of Keap1 is Cys151, which is located at the BTB domain and is flanked by the basic residues His129, Lys131, Arg135, Lys150, and His154 [51].”
BackgroundEggler, A. L., Gay, K. A., & Mesecar, A. D. (2008) · Molecular Nutrition & Food Research · doi:10.1002/mnfr.200700249 · Cited by 234 · see citing works
“2A) that mediate binding to the Keap1 Kelch repeats [20, 21].”
Ogura, T., Tong, K. I., Mio, K., Maruyama, Y., Kurokawa, H., Sato, C., & Yamamoto, M. P. (2010) · Proceedings of the National Academy of Sciences · doi:10.1073/pnas.0914036107 · Cited by 232 · see citing works
“The NMR analysis further revealed that the α-helix is mostly rigid, whereas the region from the N-terminal to the α-helix is rather flexible (27).”
BackgroundMethodInfluentialSelf-citationThe rise of antioxidant signaling—The evolution and hormetic actions of Nrf2
Maher, J. M., & Yamamoto, M. P. (2010) · Toxicology and Applied Pharmacology · doi:10.1016/j.taap.2010.01.011 · Cited by 231 · see citing works
Self-citationThe Keap1-Nrf2 system and diabetes mellitus
Uruno, A., Yagishita, Y., & Yamamoto, M. (2015) · Archives of Biochemistry and Biophysics · doi:10.1016/j.abb.2014.12.012 · Cited by 231 · see citing works
Antioxidants in Translational Medicine
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“Adduct formation with electrophiles induces a conformational change in KEAP1 that prevents further degradation of NRF2 (145).”
BackgroundStructural assembly of cullin-RING ubiquitin ligase complexes
Zimmerman, E. S., Schulman, B. A., & Zheng, N. (2010) · Current Opinion in Structural Biology · doi:10.1016/j.sbi.2010.08.010 · Cited by 227 · see citing works
Mechanism of the Nrf2/Keap1/ARE signaling system
Tkachev, V., Брониславовна, М. Е., & Зенков, Н. К. (2011) · Biochemistry (Moscow) · doi:10.1134/s0006297911040031 · Cited by 228 · see citing works
- Authors
- Tong, K. I., Katoh, Y., Kusunoki, H., Itoh, K., Tanaka, T., & Yamamoto, M.
- Affiliations
- University of Tsukuba (JP)
- Corresponding author
- Tanaka, Toshiyuki
- Yamamoto, Masayuki
- Venue
- Molecular and Cellular Biology
- Publisher
- Informa UK Limited
- Published
- 2006-04-01
- Volume
- 26
- Issue
- 8
- Pages
- 2887-2900
- ISSN
- 0270-7306 (ISSN-L)
- 1098-5549
- PMC
- PMC1446969
- OpenAlex
- W2171218310
- Cited by
- 759
- References
- 71
- Citations by year
- 2026: 44 · 2025: 44 · 2024: 42 · 2023: 56 · 2022: 58 · 2021: 46 · 2020: 59 · 2019: 41 · 2018: 38 · 2017: 51 · 2016: 31 · 2015: 34 · 2014: 36 · 2013: 35 · 2012: 30
- FWCI
- 7.06
- Type
- journal article
- Language
- en
- Open access
- green
- europepmc.org
- Locations
- Molecular and Cellular Biology (publishedVersion)
- Europe PMC (PubMed Central) (publishedVersion)
- PubMed Central (publishedVersion)
- PubMed (publishedVersion)
- Topics
- Life Sciences › Biochemistry, Genetics and Molecular Biology › Molecular Biology › Genomics, phytochemicals, and oxidative stress
- Life Sciences › Biochemistry, Genetics and Molecular Biology › Molecular Biology › Protein Structure and Dynamics
- Life Sciences › Biochemistry, Genetics and Molecular Biology › Molecular Biology › Glutathione Transferases and Polymorphisms
- Keywords
- Keap1, Nrf2, protein-protein interaction, two-site model, beta-propeller, intrinsically disordered proteins, ubiquitin ligase, post-translational regulation, antioxidant proteins
- MeSH terms
- Kelch-Like ECH-Associated Protein 1, Amino Acid Sequence, Animals, Binding Sites, Calorimetry, Cytoskeletal Proteins, Escherichia coli, Models, Chemical, Models, Molecular, Molecular Sequence Data, Protein Binding, Protein Processing, Post-Translational, Recombinant Fusion Proteins, Thermodynamics, Ultracentrifugation, Conserved Sequence, Point Mutation, Sequence Homology, Amino Acid, Protein Structure, Secondary, Protein Structure, Tertiary, Nuclear Magnetic Resonance, Biomolecular, Amino Acid Motifs, Adaptor Proteins, Signal Transducing, NF-E2-Related Factor 2, Mice
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