The Inhibitory Effect of Magnolol on the Human TWIK1 Channel Is Related to G229 and T225 Sites
TWIK1 (K2P1.1/KCNK1) belongs to the potassium channels of the two-pore domain. Its current is very small and difficult to measure. In this work, we used a 100 mM NH<sub>4</sub><sup>+</sup> extracellular solution to increase TWIK1 current in its stable cell line expressed in H...
| Main Authors: | , , , , , , , , , |
|---|---|
| Format: | Article |
| Language: | English |
| Published: |
MDPI AG
2023-09-01
|
| Series: | Molecules |
| Subjects: | |
| Online Access: | https://www.mdpi.com/1420-3049/28/19/6815 |
| _version_ | 1797575448211750912 |
|---|---|
| author | Jintao Wang Huan Liu Zhuolin Sun Xinyi Zou Zixuan Zhang Xiaofeng Wei Lanying Pan Antony Stalin Wei Zhao Yuan Chen |
| author_facet | Jintao Wang Huan Liu Zhuolin Sun Xinyi Zou Zixuan Zhang Xiaofeng Wei Lanying Pan Antony Stalin Wei Zhao Yuan Chen |
| author_sort | Jintao Wang |
| collection | DOAJ |
| description | TWIK1 (K2P1.1/KCNK1) belongs to the potassium channels of the two-pore domain. Its current is very small and difficult to measure. In this work, we used a 100 mM NH<sub>4</sub><sup>+</sup> extracellular solution to increase TWIK1 current in its stable cell line expressed in HEK293. Then, the inhibition of magnolol on TWIK1 was observed via a whole-cell patch clamp experiment, and it was found that magnolol had a significant inhibitory effect on TWIK1 (IC<sub>50</sub> = 6.21 ± 0.13 μM). By molecular docking and alanine scanning mutagenesis, the IC<sub>50</sub> of TWIK1 mutants G229A, T225A, I140A, L223A, and S224A was 20.77 ± 3.20, 21.81 ± 7.93, 10.22 ± 1.07, 9.55 ± 1.62, and 7.43 ± 3.20 μM, respectively. Thus, we conclude that the inhibition of the TWIK1 channel by magnolol is related to G229 and T225 on the P2- pore helix. |
| first_indexed | 2024-03-10T21:39:48Z |
| format | Article |
| id | doaj.art-6cc33344d8cb4077a31880f82a816440 |
| institution | Directory Open Access Journal |
| issn | 1420-3049 |
| language | English |
| last_indexed | 2024-03-10T21:39:48Z |
| publishDate | 2023-09-01 |
| publisher | MDPI AG |
| record_format | Article |
| series | Molecules |
| spelling | doaj.art-6cc33344d8cb4077a31880f82a8164402023-11-19T14:45:57ZengMDPI AGMolecules1420-30492023-09-012819681510.3390/molecules28196815The Inhibitory Effect of Magnolol on the Human TWIK1 Channel Is Related to G229 and T225 SitesJintao Wang0Huan Liu1Zhuolin Sun2Xinyi Zou3Zixuan Zhang4Xiaofeng Wei5Lanying Pan6Antony Stalin7Wei Zhao8Yuan Chen9Zhejiang Provincial Key Laboratory of Resources Protection and Innovation of Traditional Chinese Medicine, College of Food and Health, Zhejiang Agriculture and Forestry University, Hangzhou 311300, ChinaZhejiang Provincial Key Laboratory of Resources Protection and Innovation of Traditional Chinese Medicine, College of Food and Health, Zhejiang Agriculture and Forestry University, Hangzhou 311300, ChinaZhejiang Provincial Key Laboratory of Resources Protection and Innovation of Traditional Chinese Medicine, College of Food and Health, Zhejiang Agriculture and Forestry University, Hangzhou 311300, ChinaZhejiang Provincial Key Laboratory of Resources Protection and Innovation of Traditional Chinese Medicine, College of Food and Health, Zhejiang Agriculture and Forestry University, Hangzhou 311300, ChinaZhejiang Provincial Key Laboratory of Resources Protection and Innovation of Traditional Chinese Medicine, College of Food and Health, Zhejiang Agriculture and Forestry University, Hangzhou 311300, ChinaZhejiang Provincial Key Laboratory of Resources Protection and Innovation of Traditional Chinese Medicine, College of Food and Health, Zhejiang Agriculture and Forestry University, Hangzhou 311300, ChinaShulan International Medical College, Zhejiang Shuren University, Hangzhou 310015, ChinaInstitute of Fundamental and Frontier Sciences, University of Electronic Science and Technology of China, Chengdu 610054, ChinaZhejiang Provincial Key Laboratory of Resources Protection and Innovation of Traditional Chinese Medicine, College of Food and Health, Zhejiang Agriculture and Forestry University, Hangzhou 311300, ChinaZhejiang Provincial Key Laboratory of Resources Protection and Innovation of Traditional Chinese Medicine, College of Food and Health, Zhejiang Agriculture and Forestry University, Hangzhou 311300, ChinaTWIK1 (K2P1.1/KCNK1) belongs to the potassium channels of the two-pore domain. Its current is very small and difficult to measure. In this work, we used a 100 mM NH<sub>4</sub><sup>+</sup> extracellular solution to increase TWIK1 current in its stable cell line expressed in HEK293. Then, the inhibition of magnolol on TWIK1 was observed via a whole-cell patch clamp experiment, and it was found that magnolol had a significant inhibitory effect on TWIK1 (IC<sub>50</sub> = 6.21 ± 0.13 μM). By molecular docking and alanine scanning mutagenesis, the IC<sub>50</sub> of TWIK1 mutants G229A, T225A, I140A, L223A, and S224A was 20.77 ± 3.20, 21.81 ± 7.93, 10.22 ± 1.07, 9.55 ± 1.62, and 7.43 ± 3.20 μM, respectively. Thus, we conclude that the inhibition of the TWIK1 channel by magnolol is related to G229 and T225 on the P2- pore helix.https://www.mdpi.com/1420-3049/28/19/6815magnololtwo-pore domain potassium channelTWIK1 |
| spellingShingle | Jintao Wang Huan Liu Zhuolin Sun Xinyi Zou Zixuan Zhang Xiaofeng Wei Lanying Pan Antony Stalin Wei Zhao Yuan Chen The Inhibitory Effect of Magnolol on the Human TWIK1 Channel Is Related to G229 and T225 Sites Molecules magnolol two-pore domain potassium channel TWIK1 |
| title | The Inhibitory Effect of Magnolol on the Human TWIK1 Channel Is Related to G229 and T225 Sites |
| title_full | The Inhibitory Effect of Magnolol on the Human TWIK1 Channel Is Related to G229 and T225 Sites |
| title_fullStr | The Inhibitory Effect of Magnolol on the Human TWIK1 Channel Is Related to G229 and T225 Sites |
| title_full_unstemmed | The Inhibitory Effect of Magnolol on the Human TWIK1 Channel Is Related to G229 and T225 Sites |
| title_short | The Inhibitory Effect of Magnolol on the Human TWIK1 Channel Is Related to G229 and T225 Sites |
| title_sort | inhibitory effect of magnolol on the human twik1 channel is related to g229 and t225 sites |
| topic | magnolol two-pore domain potassium channel TWIK1 |
| url | https://www.mdpi.com/1420-3049/28/19/6815 |
| work_keys_str_mv | AT jintaowang theinhibitoryeffectofmagnololonthehumantwik1channelisrelatedtog229andt225sites AT huanliu theinhibitoryeffectofmagnololonthehumantwik1channelisrelatedtog229andt225sites AT zhuolinsun theinhibitoryeffectofmagnololonthehumantwik1channelisrelatedtog229andt225sites AT xinyizou theinhibitoryeffectofmagnololonthehumantwik1channelisrelatedtog229andt225sites AT zixuanzhang theinhibitoryeffectofmagnololonthehumantwik1channelisrelatedtog229andt225sites AT xiaofengwei theinhibitoryeffectofmagnololonthehumantwik1channelisrelatedtog229andt225sites AT lanyingpan theinhibitoryeffectofmagnololonthehumantwik1channelisrelatedtog229andt225sites AT antonystalin theinhibitoryeffectofmagnololonthehumantwik1channelisrelatedtog229andt225sites AT weizhao theinhibitoryeffectofmagnololonthehumantwik1channelisrelatedtog229andt225sites AT yuanchen theinhibitoryeffectofmagnololonthehumantwik1channelisrelatedtog229andt225sites AT jintaowang inhibitoryeffectofmagnololonthehumantwik1channelisrelatedtog229andt225sites AT huanliu inhibitoryeffectofmagnololonthehumantwik1channelisrelatedtog229andt225sites AT zhuolinsun inhibitoryeffectofmagnololonthehumantwik1channelisrelatedtog229andt225sites AT xinyizou inhibitoryeffectofmagnololonthehumantwik1channelisrelatedtog229andt225sites AT zixuanzhang inhibitoryeffectofmagnololonthehumantwik1channelisrelatedtog229andt225sites AT xiaofengwei inhibitoryeffectofmagnololonthehumantwik1channelisrelatedtog229andt225sites AT lanyingpan inhibitoryeffectofmagnololonthehumantwik1channelisrelatedtog229andt225sites AT antonystalin inhibitoryeffectofmagnololonthehumantwik1channelisrelatedtog229andt225sites AT weizhao inhibitoryeffectofmagnololonthehumantwik1channelisrelatedtog229andt225sites AT yuanchen inhibitoryeffectofmagnololonthehumantwik1channelisrelatedtog229andt225sites |