Efficient ASK‐assisted system for expression and purification of plant F‐box proteins |
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| Authors: | Haiou Li Ruifeng Yao Sui Ma Shuai Hu Suhua Li Yupei Wang Chun Yan Daoxin Xie Jianbin Yan |
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| Institution: | 1. MOE Key Laboratory of Bioinformatics, Tsinghua‐Peking Joint Center for Life Sciences, School of Life Sciences, Tsinghua University, Beijing, China;2. Hunan Provincial Key Laboratory of Phytohormones and Growth Development, Hunan Provincial Key Laboratory for Crop Germplasm Innovation and Utilization, Hunan Agricultural University, Changsha, China;3. Center for Synthetic and Systems Biology, Tsinghua University, Beijing, China |
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| Abstract: | Ubiquitin‐mediated protein degradation plays an essential role in plant growth and development as well as responses to environmental and endogenous signals. F‐box protein is one of the key components of the SCF (SKP1‐CUL1‐F‐box protein) E3 ubiquitin ligase complex, which recruit specific substrate proteins for subsequent ubiquitination and 26S proteasome‐mediated degradation to regulate developmental processes and signaling networks. However, it is not easy to obtain purified F‐box proteins with high activity due to their unstable protein structures. Here, we found that Arabidopsis SKP‐like proteins (ASKs) can significantly improve soluble expression of F‐box proteins and maintain their bioactivity. We established an efficient ASK‐assisted method to express and purify plant F‐box proteins. The method meets a broad range of criteria required for the biochemical analysis or protein crystallization of plant F‐box proteins. |
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| Keywords: | COI1 SLY1 TIR1
ASK
F‐box protein Technical Advance |
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