Academic Journal
Target protein identification in live cells and organisms with a non-diffusive proximity tagging system
| العنوان: | Target protein identification in live cells and organisms with a non-diffusive proximity tagging system |
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| المؤلفون: | Yingjie Sun, Changheng Li, Xiaofei Deng, Wenjie Li, Xiaoyi Deng, Weiqi Ge, Miaoyuan Shi, Ying Guo, Yanxun V Yu, Hai-bing Zhou, Youngnam N Jin |
| المصدر: | eLife, Vol 13 (2024) |
| بيانات النشر: | eLife Sciences Publications Ltd |
| سنة النشر: | 2024 |
| المجموعة: | Directory of Open Access Journals: DOAJ Articles |
| مصطلحات موضوعية: | proximity labeling, PafA, dasatinib, chloroquine, zebrafish, target identification, Medicine, Science, Biology (General), QH301-705.5 |
| الوصف: | Identifying target proteins for bioactive molecules is essential for understanding their mechanisms, developing improved derivatives, and minimizing off-target effects. Despite advances in target identification (target-ID) technologies, significant challenges remain, impeding drug development. Most target-ID methods use cell lysates, but maintaining an intact cellular context is vital for capturing specific drug–protein interactions, such as those with transient protein complexes and membrane-associated proteins. To address these limitations, we developed POST-IT (Pup-On-target for Small molecule Target Identification Technology), a non-diffusive proximity tagging system for live cells, orthogonal to the eukaryotic system. POST-IT utilizes an engineered fusion of proteasomal accessory factor A and HaloTag to transfer Pup to proximal proteins upon directly binding to the small molecule. After significant optimization to eliminate self-pupylation and polypupylation, minimize depupylation, and optimize chemical linkers, POST-IT successfully identified known targets and discovered a new binder, SEPHS2, for dasatinib, and VPS37C as a new target for hydroxychloroquine, enhancing our understanding these drugs’ mechanisms of action. Furthermore, we demonstrated the application of POST-IT in live zebrafish embryos, highlighting its potential for broad biological research and drug development. |
| نوع الوثيقة: | article in journal/newspaper |
| اللغة: | English |
| Relation: | https://elifesciences.org/articles/102667; https://doaj.org/toc/2050-084X; RP102667; https://doaj.org/article/e3f6f4fa770347f5a71bc78b3411499a |
| DOI: | 10.7554/eLife.102667 |
| الاتاحة: | https://doi.org/10.7554/eLife.102667 https://doaj.org/article/e3f6f4fa770347f5a71bc78b3411499a |
| رقم الانضمام: | edsbas.1B74305D |
| قاعدة البيانات: | BASE |
| DOI: | 10.7554/eLife.102667 |
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