Peptide inhibitors of the Keap1-Nrf2 protein-protein interaction

TY - JOUR

T1 - Peptide inhibitors of the Keap1-Nrf2 protein-protein interaction

A1 - Hancock,Rowena

A1 - Bertrand,Helene C.

A1 - Tsujita,Tadayuki

A1 - Naz,Shama

A1 - El-Bakry,Ayman

A1 - Laoruchupong,Jitnueng

A1 - Hayes,John D.

A1 - Wells,Geoff

AU - Hancock,Rowena

AU - Bertrand,Helene C.

AU - Tsujita,Tadayuki

AU - Naz,Shama

AU - El-Bakry,Ayman

AU - Laoruchupong,Jitnueng

AU - Hayes,John D.

AU - Wells,Geoff

PY - 2012/1/15

Y1 - 2012/1/15

N2 - <p>Disruption of the interaction between the ubiquitination facilitator protein Keap1 and the cap'n'collar basic-region leucine-zipper transcription factor Nrf2 is a potential strategy to enhance expression of antioxidant and free radical detoxification gene products regulated by Nrf2. Agents that disrupt this protein-protein interaction may be useful pharmacological probes and future cancer-chemopreventive agents. We describe the structure-activity relationships for a series of peptides based upon regions of the Nrf2 Neh2 domain, of varying length and sequence, that interact with the Keap1 Kelch domain and disrupt the interaction with Nrf2. We have also investigated sequestosome-1 (p62) and prothymosin-a sequences that have been reported to interact with Keap1. To achieve this we have developed a high-throughput fluorescence polarization (FP) assay to screen inhibitors. In addition to screening synthetic peptides, we have used a phage display library approach to identify putative peptide ligands with non-native sequence motifs. Candidate peptides from the phage display library screening protocol were evaluated in the FP assay to quantify their binding activity. Hybrid peptides based upon the Nrf2 "ETGE" motif and the sequestosome-1 "Keap1-interaction region" have superior binding activity compared to either native peptide alone. (C) 2011 Elsevier Inc. All rights reserved.</p>

AB - <p>Disruption of the interaction between the ubiquitination facilitator protein Keap1 and the cap'n'collar basic-region leucine-zipper transcription factor Nrf2 is a potential strategy to enhance expression of antioxidant and free radical detoxification gene products regulated by Nrf2. Agents that disrupt this protein-protein interaction may be useful pharmacological probes and future cancer-chemopreventive agents. We describe the structure-activity relationships for a series of peptides based upon regions of the Nrf2 Neh2 domain, of varying length and sequence, that interact with the Keap1 Kelch domain and disrupt the interaction with Nrf2. We have also investigated sequestosome-1 (p62) and prothymosin-a sequences that have been reported to interact with Keap1. To achieve this we have developed a high-throughput fluorescence polarization (FP) assay to screen inhibitors. In addition to screening synthetic peptides, we have used a phage display library approach to identify putative peptide ligands with non-native sequence motifs. Candidate peptides from the phage display library screening protocol were evaluated in the FP assay to quantify their binding activity. Hybrid peptides based upon the Nrf2 "ETGE" motif and the sequestosome-1 "Keap1-interaction region" have superior binding activity compared to either native peptide alone. (C) 2011 Elsevier Inc. All rights reserved.</p>

KW - Fluorescence polarization

KW - Cancer chemoprevention

KW - Compound screening

KW - Nrf2

KW - Keap1

KW - Free radicals

KW - TRANSCRIPTION FACTOR NRF2

KW - OXIDATIVE STRESS

KW - PROTEASOMAL DEGRADATION

KW - GENE-EXPRESSION

KW - DLG MOTIFS

KW - ACTIVATION

KW - MECHANISM

KW - PATHWAY

KW - CANCER

KW - UBIQUITINATION

U2 - 10.1016/j.freeradbiomed.2011.10.486

DO - 10.1016/j.freeradbiomed.2011.10.486

M1 - Article

JO - Free Radical Biology and Medicine

JF - Free Radical Biology and Medicine

SN - 0891-5849

IS - 2

VL - 52

SP - 444

EP - 451

ER -