Small molecule deoxynyboquinone triggers alkylation and ubiquitination of Keap1 at Cys489 on Kelch domain for Nrf2 activation and inflammatory therapy

Ke-Gang Linghu; Tian Zhang; Guang-Tao Zhang; Peng Lv; Wen-Jun Zhang; Guan-Ding Zhao; Shi-Hang Xiong; Qiu-Shuo Ma; Ming-Ming Zhao; Meiwan Chen; Yuan-Jia Hu; Chang-Sheng Zhang; Hua Yu

doi:10.1016/j.jpha.2023.07.009

Volume 14 Issue 3

Mar. 2024

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Article Navigation > Journal of Pharmaceutical Analysis > 2024 > 14(3): 401-415

Ke-Gang Linghu, Tian Zhang, Guang-Tao Zhang, Peng Lv, Wen-Jun Zhang, Guan-Ding Zhao, Shi-Hang Xiong, Qiu-Shuo Ma, Ming-Ming Zhao, Meiwan Chen, Yuan-Jia Hu, Chang-Sheng Zhang, Hua Yu. Small molecule deoxynyboquinone triggers alkylation and ubiquitination of Keap1 at Cys489 on Kelch domain for Nrf2 activation and inflammatory therapy[J]. Journal of Pharmaceutical Analysis, 2024, 14(3): 401-415. doi: 10.1016/j.jpha.2023.07.009

Citation:

Ke-Gang Linghu, Tian Zhang, Guang-Tao Zhang, Peng Lv, Wen-Jun Zhang, Guan-Ding Zhao, Shi-Hang Xiong, Qiu-Shuo Ma, Ming-Ming Zhao, Meiwan Chen, Yuan-Jia Hu, Chang-Sheng Zhang, Hua Yu. Small molecule deoxynyboquinone triggers alkylation and ubiquitination of Keap1 at Cys489 on Kelch domain for Nrf2 activation and inflammatory therapy[J]. Journal of Pharmaceutical Analysis, 2024, 14(3): 401-415. doi: 10.1016/j.jpha.2023.07.009

Citation:

Ke-Gang Linghu, Tian Zhang, Guang-Tao Zhang, Peng Lv, Wen-Jun Zhang, Guan-Ding Zhao, Shi-Hang Xiong, Qiu-Shuo Ma, Ming-Ming Zhao, Meiwan Chen, Yuan-Jia Hu, Chang-Sheng Zhang, Hua Yu. Small molecule deoxynyboquinone triggers alkylation and ubiquitination of Keap1 at Cys489 on Kelch domain for Nrf2 activation and inflammatory therapy[J]. Journal of Pharmaceutical Analysis, 2024, 14(3): 401-415. doi: 10.1016/j.jpha.2023.07.009

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Small molecule deoxynyboquinone triggers alkylation and ubiquitination of Keap1 at Cys489 on Kelch domain for Nrf2 activation and inflammatory therapy

doi: 10.1016/j.jpha.2023.07.009

a. State Key Laboratory of Quality Research in Chinese Medicine, Institute of Chinese Medical Sciences, University of Macau, Macao SAR, China;
b. Macao Centre for Research and Development in Chinese Medicine, Institute of Chinese Medical Sciences, University of Macau, Macao SAR, China;
c. Key Laboratory of Tropical Marine Bio-resources and Ecology, Guangdong Key Laboratory of Marine Materia Medica, RNAM Center for Marine Microbiology, South China Sea Institute of Oceanology, Chinese Academy of Sciences, Guangzhou, 510301, China;
d. State Key Laboratory of Functions and Applications of Medicinal Plants & School of Pharmaceutical Sciences, Guizhou Medical University, Guiyang, 550025, China

Funds:

This study was supported by the Science and Technology Development Fund, Macao SAR (Grant Nos.: No.0159/2020/A3, No.0058/2020/AMJ, No.0096/2019/A2 and SKL-QRCM(UM)-2023-2025), the Research Committee of the University of Macau (Grant No.: MYRG2022-00189-ICMS), the Guangdong Provincial Special Fund for Marine Economic Development Project (Project No.: GDNRC[2021]48), National Natural Science Foundation of China (Grant No.: 82260801), and K.C. Wong Education Foundation (Grant No.: GJTD-2020-12). We greatly appreciate Ms. Parsa Dar for her language help.

Received Date: Apr. 18, 2023
Accepted Date: Jul. 13, 2023
Rev Recd Date: Jul. 13, 2023
Publish Date: Jul. 17, 2023

Abstract

Abstract

Activation of nuclear factor erythroid 2-related factor 2 (Nrf2) by Kelch-like ECH-associated protein 1 (Keap1) alkylation plays a central role in anti-inflammatory therapy. However, activators of Nrf2 through alkylation of Keap1-Kelch domain have not been identified. Deoxynyboquinone (DNQ) is a natural small molecule discovered from marine actinomycetes. The current study was designed to investigate the anti-inflammatory effects and molecular mechanisms of DNQ via alkylation of Keap1. DNQ exhibited significant anti-inflammatory properties both in vitro and in vivo. The pharmacophore responsible for the anti-inflammatory properties of DNQ was determined to be the α, β-unsaturated amides moieties by a chemical reaction between DNQ and N-acetylcysteine. DNQ exerted anti-inflammatory effects through activation of Nrf2/ARE pathway. Keap1 was demonstrated to be the direct target of DNQ and bound with DNQ through conjugate addition reaction involving alkylation. The specific alkylation site of DNQ on Keap1 for Nrf2 activation was elucidated with a synthesized probe in conjunction with liquid chromatography-tandem mass spectrometry. DNQ triggered the ubiquitination and subsequent degradation of Keap1 by alkylation of the cysteine residue 489 (Cys489) on Keap1-Kelch domain, ultimately enabling the activation of Nrf2. Our findings revealed that DNQ exhibited potent anti-inflammatory capacity through α, β-unsaturated amides moieties active group which specifically activated Nrf2 signal pathway via alkylation/ubiquitination of Keap1-Kelch domain, suggesting the potential values of targeting Cys489 on Keap1-Kelch domain by DNQ-like small molecules in inflammatory therapies.
- Deoxynyboquinone,
- Anti-inflammation,
- Target,
- Keap1/Nrf2,
- Alkylation,
- Ubiquitination

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References(42)

References

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