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Efficient ASK ‐assisted system for expression and purification of plant F‐box proteins
Author(s) -
Li Haiou,
Yao Ruifeng,
Ma Sui,
Hu Shuai,
Li Suhua,
Wang Yupei,
Yan Chun,
Xie Daoxin,
Yan Jianbin
Publication year - 2017
Publication title -
the plant journal
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 3.058
H-Index - 269
eISSN - 1365-313X
pISSN - 0960-7412
DOI - 10.1111/tpj.13708
Subject(s) - f box protein , ubiquitin ligase , ubiquitin , proteasome , biochemistry , arabidopsis , biology , protein degradation , microbiology and biotechnology , chemistry , mutant , gene
Summary 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 ( SKP 1‐ CUL 1‐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 ( ASK s) 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.