Guiding bar motif of thioredoxin reductase 1 modulates enzymatic activity and inhibitor binding by communicating with the co-factor FAD and regulating the flexible C-terminal redox motif
Thioredoxin reductase (TXNRD) is a selenoprotein that plays a crucial role in cellular antioxidant defense. Previously, a distinctive guiding bar motif was identified in TXNRD1, which influences the transfer of electrons. In this study, utilizing single amino acid substitution and Excitation-Emissio...
| Main Authors: | , , , , , , , , , , , , , , |
|---|---|
| 格式: | 文件 |
| 语言: | English |
| 出版: |
Elsevier
2024-04-01
|
| 丛编: | Redox Biology |
| 主题: | |
| 在线阅读: | http://www.sciencedirect.com/science/article/pii/S2213231724000260 |
| _version_ | 1827345414681001984 |
|---|---|
| author | Wuyang Shi Shibo Sun Haowen Liu Yao Meng Kangshuai Ren Guoying Wang Minghui Liu Jiaqi Wu Yue Zhang Huang Huang Meiyun Shi Weiping Xu Qiang Ma Bingbing Sun Jianqiang Xu |
| author_facet | Wuyang Shi Shibo Sun Haowen Liu Yao Meng Kangshuai Ren Guoying Wang Minghui Liu Jiaqi Wu Yue Zhang Huang Huang Meiyun Shi Weiping Xu Qiang Ma Bingbing Sun Jianqiang Xu |
| author_sort | Wuyang Shi |
| collection | DOAJ |
| description | Thioredoxin reductase (TXNRD) is a selenoprotein that plays a crucial role in cellular antioxidant defense. Previously, a distinctive guiding bar motif was identified in TXNRD1, which influences the transfer of electrons. In this study, utilizing single amino acid substitution and Excitation-Emission Matrix (EEM) fluorescence spectrum analysis, we discovered that the guiding bar communicates with the FAD and modulates the electron flow of the enzyme. Differential Scanning Fluorimetry (DSF) analysis demonstrated that the aromatic amino acid in guiding bar is a stabilizer for TXNRD1. Kinetic analysis revealed that the guiding bar is vital for the disulfide reductase activity but hinders the selenocysteine-independent reduction activity of TXNRD1. Meanwhile, the guiding bar shields the selenocysteine residue of TXNRD1 from the attack of electrophilic reagents. We also found that the inhibition of TXNRD1 by caveolin-1 scaffolding domain (CSD) peptides and compound LCS3 did not bind to the guiding bar motif. In summary, the obtained results highlight new aspects of the guiding bar that restrict the flexibility of the C-terminal redox motif and govern the transition from antioxidant to pro-oxidant. |
| first_indexed | 2024-03-07T23:06:21Z |
| format | Article |
| id | doaj.art-a794bd0d20ef4b188abb9772e7ef29ce |
| institution | Directory Open Access Journal |
| issn | 2213-2317 |
| language | English |
| last_indexed | 2024-03-07T23:06:21Z |
| publishDate | 2024-04-01 |
| publisher | Elsevier |
| record_format | Article |
| series | Redox Biology |
| spelling | doaj.art-a794bd0d20ef4b188abb9772e7ef29ce2024-02-22T04:52:17ZengElsevierRedox Biology2213-23172024-04-0170103050Guiding bar motif of thioredoxin reductase 1 modulates enzymatic activity and inhibitor binding by communicating with the co-factor FAD and regulating the flexible C-terminal redox motifWuyang Shi0Shibo Sun1Haowen Liu2Yao Meng3Kangshuai Ren4Guoying Wang5Minghui Liu6Jiaqi Wu7Yue Zhang8Huang Huang9Meiyun Shi10Weiping Xu11Qiang Ma12Bingbing Sun13Jianqiang Xu14School of Life and Pharmaceutical Sciences (LPS) & Panjin Institute of Industrial Technology (PIIT), Dalian University of Technology, Panjin, 124221, ChinaSchool of Life and Pharmaceutical Sciences (LPS) & Panjin Institute of Industrial Technology (PIIT), Dalian University of Technology, Panjin, 124221, ChinaSchool of Life and Pharmaceutical Sciences (LPS) & Panjin Institute of Industrial Technology (PIIT), Dalian University of Technology, Panjin, 124221, ChinaSchool of Life and Pharmaceutical Sciences (LPS) & Panjin Institute of Industrial Technology (PIIT), Dalian University of Technology, Panjin, 124221, ChinaSchool of Life and Pharmaceutical Sciences (LPS) & Panjin Institute of Industrial Technology (PIIT), Dalian University of Technology, Panjin, 124221, ChinaSchool of Life and Pharmaceutical Sciences (LPS) & Panjin Institute of Industrial Technology (PIIT), Dalian University of Technology, Panjin, 124221, ChinaSchool of Life and Pharmaceutical Sciences (LPS) & Panjin Institute of Industrial Technology (PIIT), Dalian University of Technology, Panjin, 124221, ChinaSchool of Life and Pharmaceutical Sciences (LPS) & Panjin Institute of Industrial Technology (PIIT), Dalian University of Technology, Panjin, 124221, ChinaSchool of Life and Pharmaceutical Sciences (LPS) & Panjin Institute of Industrial Technology (PIIT), Dalian University of Technology, Panjin, 124221, ChinaSchool of Life and Pharmaceutical Sciences (LPS) & Panjin Institute of Industrial Technology (PIIT), Dalian University of Technology, Panjin, 124221, ChinaSchool of Life and Pharmaceutical Sciences (LPS) & Panjin Institute of Industrial Technology (PIIT), Dalian University of Technology, Panjin, 124221, ChinaSchool of Ocean Science and Technology (OST) & Key Laboratory of Industrial Ecology and Environmental Engineering (Ministry of Education), Dalian University of Technology, Panjin, 124221, ChinaChinese Academy of Inspection and Quarantine, Beijing, 100176, ChinaState Key Laboratory of Fine Chemicals, School of Chemical Engineering (CE), Dalian University of Technology, Dalian, 116023, ChinaSchool of Life and Pharmaceutical Sciences (LPS) & Panjin Institute of Industrial Technology (PIIT), Dalian University of Technology, Panjin, 124221, China; Corresponding author.Thioredoxin reductase (TXNRD) is a selenoprotein that plays a crucial role in cellular antioxidant defense. Previously, a distinctive guiding bar motif was identified in TXNRD1, which influences the transfer of electrons. In this study, utilizing single amino acid substitution and Excitation-Emission Matrix (EEM) fluorescence spectrum analysis, we discovered that the guiding bar communicates with the FAD and modulates the electron flow of the enzyme. Differential Scanning Fluorimetry (DSF) analysis demonstrated that the aromatic amino acid in guiding bar is a stabilizer for TXNRD1. Kinetic analysis revealed that the guiding bar is vital for the disulfide reductase activity but hinders the selenocysteine-independent reduction activity of TXNRD1. Meanwhile, the guiding bar shields the selenocysteine residue of TXNRD1 from the attack of electrophilic reagents. We also found that the inhibition of TXNRD1 by caveolin-1 scaffolding domain (CSD) peptides and compound LCS3 did not bind to the guiding bar motif. In summary, the obtained results highlight new aspects of the guiding bar that restrict the flexibility of the C-terminal redox motif and govern the transition from antioxidant to pro-oxidant.http://www.sciencedirect.com/science/article/pii/S2213231724000260Thioredoxin reductaseThioredoxinSelenoproteinGuiding bar motifCaveolin-1LCS3 |
| spellingShingle | Wuyang Shi Shibo Sun Haowen Liu Yao Meng Kangshuai Ren Guoying Wang Minghui Liu Jiaqi Wu Yue Zhang Huang Huang Meiyun Shi Weiping Xu Qiang Ma Bingbing Sun Jianqiang Xu Guiding bar motif of thioredoxin reductase 1 modulates enzymatic activity and inhibitor binding by communicating with the co-factor FAD and regulating the flexible C-terminal redox motif Redox Biology Thioredoxin reductase Thioredoxin Selenoprotein Guiding bar motif Caveolin-1 LCS3 |
| title | Guiding bar motif of thioredoxin reductase 1 modulates enzymatic activity and inhibitor binding by communicating with the co-factor FAD and regulating the flexible C-terminal redox motif |
| title_full | Guiding bar motif of thioredoxin reductase 1 modulates enzymatic activity and inhibitor binding by communicating with the co-factor FAD and regulating the flexible C-terminal redox motif |
| title_fullStr | Guiding bar motif of thioredoxin reductase 1 modulates enzymatic activity and inhibitor binding by communicating with the co-factor FAD and regulating the flexible C-terminal redox motif |
| title_full_unstemmed | Guiding bar motif of thioredoxin reductase 1 modulates enzymatic activity and inhibitor binding by communicating with the co-factor FAD and regulating the flexible C-terminal redox motif |
| title_short | Guiding bar motif of thioredoxin reductase 1 modulates enzymatic activity and inhibitor binding by communicating with the co-factor FAD and regulating the flexible C-terminal redox motif |
| title_sort | guiding bar motif of thioredoxin reductase 1 modulates enzymatic activity and inhibitor binding by communicating with the co factor fad and regulating the flexible c terminal redox motif |
| topic | Thioredoxin reductase Thioredoxin Selenoprotein Guiding bar motif Caveolin-1 LCS3 |
| url | http://www.sciencedirect.com/science/article/pii/S2213231724000260 |
| work_keys_str_mv | AT wuyangshi guidingbarmotifofthioredoxinreductase1modulatesenzymaticactivityandinhibitorbindingbycommunicatingwiththecofactorfadandregulatingtheflexiblecterminalredoxmotif AT shibosun guidingbarmotifofthioredoxinreductase1modulatesenzymaticactivityandinhibitorbindingbycommunicatingwiththecofactorfadandregulatingtheflexiblecterminalredoxmotif AT haowenliu guidingbarmotifofthioredoxinreductase1modulatesenzymaticactivityandinhibitorbindingbycommunicatingwiththecofactorfadandregulatingtheflexiblecterminalredoxmotif AT yaomeng guidingbarmotifofthioredoxinreductase1modulatesenzymaticactivityandinhibitorbindingbycommunicatingwiththecofactorfadandregulatingtheflexiblecterminalredoxmotif AT kangshuairen guidingbarmotifofthioredoxinreductase1modulatesenzymaticactivityandinhibitorbindingbycommunicatingwiththecofactorfadandregulatingtheflexiblecterminalredoxmotif AT guoyingwang guidingbarmotifofthioredoxinreductase1modulatesenzymaticactivityandinhibitorbindingbycommunicatingwiththecofactorfadandregulatingtheflexiblecterminalredoxmotif AT minghuiliu guidingbarmotifofthioredoxinreductase1modulatesenzymaticactivityandinhibitorbindingbycommunicatingwiththecofactorfadandregulatingtheflexiblecterminalredoxmotif AT jiaqiwu guidingbarmotifofthioredoxinreductase1modulatesenzymaticactivityandinhibitorbindingbycommunicatingwiththecofactorfadandregulatingtheflexiblecterminalredoxmotif AT yuezhang guidingbarmotifofthioredoxinreductase1modulatesenzymaticactivityandinhibitorbindingbycommunicatingwiththecofactorfadandregulatingtheflexiblecterminalredoxmotif AT huanghuang guidingbarmotifofthioredoxinreductase1modulatesenzymaticactivityandinhibitorbindingbycommunicatingwiththecofactorfadandregulatingtheflexiblecterminalredoxmotif AT meiyunshi guidingbarmotifofthioredoxinreductase1modulatesenzymaticactivityandinhibitorbindingbycommunicatingwiththecofactorfadandregulatingtheflexiblecterminalredoxmotif AT weipingxu guidingbarmotifofthioredoxinreductase1modulatesenzymaticactivityandinhibitorbindingbycommunicatingwiththecofactorfadandregulatingtheflexiblecterminalredoxmotif AT qiangma guidingbarmotifofthioredoxinreductase1modulatesenzymaticactivityandinhibitorbindingbycommunicatingwiththecofactorfadandregulatingtheflexiblecterminalredoxmotif AT bingbingsun guidingbarmotifofthioredoxinreductase1modulatesenzymaticactivityandinhibitorbindingbycommunicatingwiththecofactorfadandregulatingtheflexiblecterminalredoxmotif AT jianqiangxu guidingbarmotifofthioredoxinreductase1modulatesenzymaticactivityandinhibitorbindingbycommunicatingwiththecofactorfadandregulatingtheflexiblecterminalredoxmotif |