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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 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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Cites
NMRPipe: A multidimensional spectral processing system based on UNIX pipes
Delaglio, F., Grzesiek, S., Vuister, G. W., Zhu, G., Pfeifer, J., & Bax, A. (1995) · Journal of Biomolecular NMR · doi:10.1007/bf00197809 · Cited by 15,961
NMR with Proteins and Nucleic Acids
Wüthrich, K. (1986) · Europhysics news · doi:10.1051/epn/19861701011 · Cited by 8,028
NMR of Proteins and Nucleic Acids
(2006) · Royal Society of Chemistry eBooks · doi:10.1039/9781847555236-00336 · Cited by 5,874
Investigation of exchange processes by two-dimensional NMR spectroscopy
Jeener, J., Meier, B. H., Bachmann, P., & Ernst, R. R. (1979) · The Journal of Chemical Physics · doi:10.1063/1.438208 · Cited by 4,750
Mitochondria, Oxidants, and Aging
Balaban, R. S., Nemoto, S., & Finkel, T. (2005) · Cell · doi:10.1016/j.cell.2005.02.001 · Cited by 4,388
Intrinsically unstructured proteins and their functions
Dyson, H. J., & Wright, P. E. (2005) · Nature Reviews Molecular Cell Biology · doi:10.1038/nrm1589 · Cited by 3,871
Itoh, K., Wakabayashi, N., Katoh, Y., Ishii, T., Igarashi, K., Engel, J. D., & Yamamoto, M. (1999) · Genes & Development · doi:10.1101/gad.13.1.76 · Cited by 3,634
Self-citationHayes, J. D., Flanagan, J. U., & Jowsey, I. R. (2004) · The Annual Review of Pharmacology and Toxicology · doi:10.1146/annurev.pharmtox.45.120403.095857 · Cited by 3,565
Pervushin, K. V., Riek, R., Wider, G., & Kurt Wüthrich, A. (1997) · Proceedings of the National Academy of Sciences · doi:10.1073/pnas.94.23.12366 · Cited by 2,450
Kay, L. E., Keifer, P. A., & Saarinen, T. (1992) · Journal of the American Chemical Society · doi:10.1021/ja00052a088 · Cited by 2,261
Kobayashi, A., Kang, M.-I., Okawa, H., Ohtsuji, M., Zenke, Y., Chiba, T., Igarashi, K., & Yamamoto, M. (2004) · Molecular and Cellular Biology · doi:10.1128/mcb.24.16.7130-7139.2004 · Cited by 2,224
Function and regulation of cullin–RING ubiquitin ligases
Petroski, M. D., & Deshaies, R. J. (2005) · Nature Reviews Molecular Cell Biology · doi:10.1038/nrm1547 · Cited by 2,148
Farrow, N. A., Muhandiram, R., Singer, A. U., Pascal, S. M., Kay, C. M., Gish, G., Shoelson, S. E., Pawson, T. J., Forman-Kay, J. D., & Kay, L. E. (1994) · Biochemistry · doi:10.1021/bi00185a040 · Cited by 2,094
Dinkova-Kostova, A. T., Holtzclaw, W. D., Cole, R. N., Itoh, K., Wakabayashi, N., Katoh, Y., Yamamoto, M., & Talalay, P. (2002) · Proceedings of the National Academy of Sciences · doi:10.1073/pnas.172398899 · Cited by 1,947
Self-citationKay, L. E., Torchia, D. A., & Bax, A. (1989) · Biochemistry · doi:10.1021/bi00449a003 · Cited by 1,787
Nrf2–Keap1 defines a physiologically important stress response mechanism
Motohashi, H., & Yamamoto, M. (2004) · Trends in Molecular Medicine · doi:10.1016/j.molmed.2004.09.003 · Cited by 1,725 · cited 1×
“The induction of these cytoprotective genes is regulated at the transcriptional level by a specific cis-acting element, the antioxidant/electrophile response element (ARE/EpRE) found in the promoter regions of these genes (21, 39, 40).”
BackgroundSelf-citationSattler, M. (1999) · Progress in Nuclear Magnetic Resonance Spectroscopy · doi:10.1016/s0079-6565(98)00025-9 · Cited by 1,721
Wishart, D. S., Bigam, C. G., Holm, A., Hodges, R. S., & Sykes, B. D. (1995) · Journal of Biomolecular NMR · doi:10.1007/bf00227471 · Cited by 1,635
Moi, P., Chan, K., Asunis, I., Cao, A., & Kan, Y. W. (1994) · Proceedings of the National Academy of Sciences · doi:10.1073/pnas.91.21.9926 · Cited by 1,607
Structure of the Cul1–Rbx1–Skp1–F boxSkp2 SCF ubiquitin ligase complex
Zheng, N., Schulman, B. A., Song, L., Miller, J. J., Jeffrey, P. D., Wang, P., Chu, C., Koepp, D. M., Elledge, S. J., Pagano, M., et al. (2002) · Nature · doi:10.1038/416703a · Cited by 1,507
Keap1 Is a Redox-Regulated Substrate Adaptor Protein for a Cul3-Dependent Ubiquitin Ligase Complex
Zhang, D. D., Lo, S., Cross, J. V., Templeton, D. J., & Hannink, M. (2004) · Molecular and Cellular Biology · doi:10.1128/mcb.24.24.10941-10953.2004 · Cited by 1,284
Regulatory Mechanisms Controlling Gene Expression Mediated by the Antioxidant Response Element
Nguyen, T., Sherratt, P. J., & Pickett, C. B. (2003) · The Annual Review of Pharmacology and Toxicology · doi:10.1146/annurev.pharmtox.43.100901.140229 · Cited by 1,232
Wishart, D. S., Bigam, C. G., Holm, A., Hodges, R. S., & Sykes, B. D. (1995) · Journal of Biomolecular NMR · doi:10.1007/bf00211764 · Cited by 1,012
Wakabayashi, N., Dinkova-Kostova, A. T., Holtzclaw, W. D., Kang, M.-I., Kobayashi, A., Yamamoto, M., Kensler, T. W., & Talalay, P. (2004) · Proceedings of the National Academy of Sciences · doi:10.1073/pnas.0307301101 · Cited by 986
Cullinan, S. B., Gordan, J. D., Jin, J., Harper, J. W., & Diehl, J. A. (2004) · Molecular and Cellular Biology · doi:10.1128/mcb.24.19.8477-8486.2004 · Cited by 986
Pervushin, K. V., Riek, R., Wider, G., & Kurt Wüthrich, A. (2021) · WORLD SCIENTIFIC eBooks · doi:10.1142/9789811235795_0006 · Cited by 951
Kobayashi, A., Kang, M.-I., Watai, Y., Tong, K. I., Shibata, T., Uchida, K., & Yamamoto, M. P. (2005) · Molecular and Cellular Biology · doi:10.1128/mcb.26.1.221-229.2006 · Cited by 947
Self-citationGradient-Enhanced Triple-Resonance Three-Dimensional NMR Experiments with Improved Sensitivity
Muhandiram, D. R., & Kay, L. E. (1994) · Journal of Magnetic Resonance Series B · doi:10.1006/jmrb.1994.1032 · Cited by 901
Itoh, K., Tong, K. I., & Yamamoto, M. P. (2004) · Free Radical Biology and Medicine · doi:10.1016/j.freeradbiomed.2004.02.075 · Cited by 873
Self-citationAnalysis of data from the analytical ultracentrifuge by nonlinear least-squares techniques
Johnson, M. L., Correia, J. J., Yphantis, D. A., & Halvorson, H. R. (1981) · Biophysical Journal · doi:10.1016/s0006-3495(81)84753-4 · Cited by 844
Furukawa, M., & Xiong, Y. (2004) · Molecular and Cellular Biology · doi:10.1128/mcb.25.1.162-171.2005 · Cited by 759
The POZ domain: a conserved protein-protein interaction motif.Preprint
Bardwell, V. J., & Treisman, R. (1994) · Genes & Development · doi:10.1101/gad.8.14.1664 · Cited by 749
Structural basis for defects of Keap1 activity provoked by its point mutations in lung cancer
Padmanabhan, B., Tong, K. I., Ohta, T., Nakamura, Y., Scharlock, M., Ohtsuji, M., Kang, M.-I., Kobayashi, A., Yokoyama, S., & Yamamoto, M. (2006) · Molecular Cell · doi:10.1016/j.molcel.2006.01.013 · Cited by 735
Self-citationThe interplay between structure and function in intrinsically unstructured proteins
Tompa, P. (2005) · FEBS Letters · doi:10.1016/j.febslet.2005.03.072 · Cited by 717
Sequence and structural analysis of BTB domain proteins
Stogios, P. J., Downs, G. S., Jauhal, J. J., Nandra, S. K., & Privé, G. G. (2005) · Genome Biology · doi:10.1186/gb-2005-6-10-r82 · Cited by 694
Garrett, D. S., Powers, R., Gronenborn, A. M., & Clore, G. M. (1991) · Journal of Magnetic Resonance (1969) · doi:10.1016/0022-2364(91)90341-p · Cited by 682
Zhang, O., Kay, L. E., Olivier, J. P., & Forman-Kay, J. D. (1994) · Journal of Biomolecular NMR · doi:10.1007/bf00398413 · Cited by 648
The kelch repeat superfamily of proteins: propellers of cell function
Adams, J. C., Kelso, R., & Cooley, L. (2000) · Trends in Cell Biology · doi:10.1016/s0962-8924(99)01673-6 · Cited by 603
Katoh, Y., Itoh, K., Yoshida, E., Miyagishi, M., Fukamizu, A., & Yamamoto, M. P. (2001) · Genes to Cells · doi:10.1046/j.1365-2443.2001.00469.x · Cited by 533
Self-citationOrlicky, S. M., Tang, X., Willems, A. R., Tyers, M., & Sicheri, F. (2003) · Cell · doi:10.1016/s0092-8674(03)00034-5 · Cited by 515
The pathways and molecular mechanisms regulating Nrf2 activation in response to chemical stress
Nguyen, T., Yang, C. S., & Pickett, C. B. (2004) · Free Radical Biology and Medicine · doi:10.1016/j.freeradbiomed.2004.04.033 · Cited by 472
Motohashi, H., O'Connor, T., Katsuoka, F., Engel, J. D., & Yamamoto, M. P. (2002) · Gene · doi:10.1016/s0378-1119(02)00788-6 · Cited by 456
Self-citationKelch encodes a component of intercellular bridges in Drosophila egg chambers
Xue, F. (1993) · Cell · doi:10.1016/0092-8674(93)90397-9 · Cited by 440
Itoh, K., Igarashi, K., Hayashi, N., Nishizawa, M., & Yamamoto, M. P. (1995) · Molecular and Cellular Biology · doi:10.1128/mcb.15.8.4184 · Cited by 438
Self-citationCullin‐based ubiquitin ligases: Cul3–BTB complexes join the family
Pintard, L., Willems, A. R., & Peter, M. (2004) · The EMBO Journal · doi:10.1038/sj.emboj.7600186 · Cited by 422
McMahon, M. J., Thomas, N., Itoh, K., Yamamoto, M. P., & Hayes, J. D. (2004) · Journal of Biological Chemistry · doi:10.1074/jbc.m403061200 · Cited by 419
Self-citationThe Keap1 BTB/POZ dimerization function is required to sequester Nrf2 in cytoplasm
Zipper, L. M., & Mulcahy, R. T. (2002) · Journal of Biological Chemistry · doi:10.1074/jbc.m206530200 · Cited by 366
Nioi, P., & Hayes, J. D. (2004) · Mutation research. Fundamental and molecular mechanisms of mutagenesis · doi:10.1016/j.mrfmmm.2004.05.023 · Cited by 364
Targeted ubiquitination of CDT1 by the DDB1–CUL4A–ROC1 ligase in response to DNA damage
Hu, J., McCall, C. M., Ohta, T., & Xiong, Y. (2004) · Nature Cell Biology · doi:10.1038/ncb1172 · Cited by 352
Identification of the interactive interface and phylogenic conservation of the Nrf2‐Keap1 system
Kobayashi, M., Itoh, K., Suzuki, T., Osanai, H., Nishikawa, K., Katoh, Y., Takagi, Y., & Yamamoto, M. P. (2002) · Genes to Cells · doi:10.1046/j.1365-2443.2002.00561.x · Cited by 340
Self-citationBoundary analysis of sedimentation‐velocity experiments with monodisperse and paucidisperse solutes
Holde, K. E. V., & Weischet, W. O. (1978) · Biopolymers · doi:10.1002/bip.1978.360170602 · Cited by 314
Lacy, E. R., Filippov, I. V., Lewis, W. S., Otieno, S. B., Xiao, L., Weiß, S. B., Hengst, L., & Kriwacki, R. W. (2004) · Nature Structural & Molecular Biology · doi:10.1038/nsmb746 · Cited by 305
Kay, L. E. (1998) · Nature Structural Biology · doi:10.1038/755 · Cited by 284
Crystal Structure of the Kelch Domain of Human Keap1
Li, X., Zhang, D., Hannink, M., & Beamer, L. J. (2004) · Journal of Biological Chemistry · doi:10.1074/jbc.m410073200 · Cited by 256
CYTOCHROME P450 ACTIVATION OF ARYLAMINES AND HETEROCYCLIC AMINES
Kim, D., & Guengerich, F. P. (2004) · The Annual Review of Pharmacology and Toxicology · doi:10.1146/annurev.pharmtox.45.120403.100010 · Cited by 250
Dietary chemopreventive compounds and ARE/EpRE signaling
Chen, C., & Kong, A. (2004) · Free Radical Biology and Medicine · doi:10.1016/j.freeradbiomed.2004.03.015 · Cited by 241
β Propellers: structural rigidity and functional diversity
Fülöp, V., & Jones, D. T. (1999) · Current Opinion in Structural Biology · doi:10.1016/s0959-440x(99)00035-4 · Cited by 232
Katoh, Y., Iida, K., Kang, M.-I., Kobayashi, A., Mizukami, M., Tong, K. I., McMahon, M. J., Hayes, J. D., Itoh, K., & Yamamoto, M. P. (2004) · Archives of Biochemistry and Biophysics · doi:10.1016/j.abb.2004.10.012 · Cited by 217
Self-citationHoltzclaw, W. D., Dinkova‐Kostova, A. T., & Talalay, P. (2004) · Advances in Enzyme Regulation · doi:10.1016/j.advenzreg.2003.11.013 · Cited by 145
Garrett, D. S., Powers, R., Gronenborn, A. M., & Clore, G. M. (2011) · Journal of Magnetic Resonance · doi:10.1016/j.jmr.2011.09.007 · Cited by 139
Metzler, W. J., Constantine, K. L., Friedrichs, M. S., Bell, A., Ernst, E., Lavoie, T., & Mueller, L. (1993) · Biochemistry · doi:10.1021/bi00213a010 · Cited by 123
F2-isoprostanes as indices of lipid peroxidation in inflammatory diseases
Praticò, D., Rokach, J., Lawson, J., & FitzGerald, G. A. (2004) · Chemistry and Physics of Lipids · doi:10.1016/j.chemphyslip.2003.09.012 · Cited by 120
Metzler, W. J., Constantine, K. L., Friedrichs, M. S., Bell, A., Ernst, E., Lavoie, T., & Mueller, L. (2001) · Europe PMC (PubMed Central) · doi:10.13018/bmr4082 · Cited by 118
Oxidative stress and atherosclerosis
Schulze, P. C., & Lee, R. T. (2005) · Current Atherosclerosis Reports · doi:10.1007/s11883-005-0013-5 · Cited by 112
Smad3–ATF3 signaling mediates TGF-β suppression of genes encoding Phase II detoxifying proteins
Bakin, A. V., Stourman, N. V., Sekhar, K. R., Rinehart, C., Yan, X. S., Meredith, M. J., Arteaga, C. L., & Freeman, M. L. (2004) · Free Radical Biology and Medicine · doi:10.1016/j.freeradbiomed.2004.10.033 · Cited by 97
Lee, A. L., & Wand, A. J. (1999) · Journal of Biomolecular NMR · doi:10.1023/a:1008304220445 · Cited by 77
Duggan, B. M., Dyson, H. J., & Wright, P. E. (1999) · European Journal of Biochemistry · doi:10.1046/j.1432-1327.1999.00781.x · Cited by 47
Ortega, Á., & de la Torre, J. G. (2005) · Journal of the American Chemical Society · doi:10.1021/ja053080l · Cited by 29
Padmanabhan, B., Scharlock, M., Tong, K. I., Nakamura, Y., Kang, M.-I., Kobayashi, A., Matsumoto, T., Tanaka, A., Yamamoto, M., & Yokoyama, S. (2004) · Acta Crystallographica Section F Structural Biology and Crystallization Communications · doi:10.1107/s1744309104032506 · Cited by 16
Self-citationIntroduction to serial reviews on EpRE and its signaling pathway
Forman, H. J., & Ruden, D. M. (2004) · Free Radical Biology and Medicine · doi:10.1016/j.freeradbiomed.2004.03.004 · Cited by 7
Research works printed but not linked: 1
- Garrett D. S., 1991, J. Magn. Reson., 95, 214
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Metadata
- 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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