Escherichia coli proteome microarrays identified the substrates of clpyq protease

Chih Hsuan Tsai, Yu Hsuan Ho, Tzu Cheng Sung, Whei Fen Wu, Chien Sheng Chen

研究成果: Article同行評審

15 引文 斯高帕斯(Scopus)


Proteolysis is a vital mechanism to regulate the cellular proteome in all kingdoms of life, and ATP-dependent proteases play a crucial role within this process. In Escherichia coli, ClpYQ is one of the primary ATP-dependent proteases. In addition to function with removals of abnormal peptides in the cells, ClpYQ degrades regulatory proteins if necessary and thus let cells adjust to various environmental conditions. In E. coli, SulA, RcsA, RpoH and TraJ as well as RNase R, have been identified as natural protein substrates of ClpYQ. ClpYQ contains ClpY and ClpQ. The ATPase ClpY is responsible for protein recognition, unfolding, and translocation into the catalytic core of ClpQ. In this study, we use an indirect identification strategy to screen possible ClpY targets with E. coli K12 proteome chips. The chip assay results showed that YbaB strongly bound to ClpY. We used yeast two-hybrid assay to confirm the interactions between ClpY and YbaB protein and determined the Kd between ClpY and YbaB by quartz crystal microbalance. Furthermore, we validated that YbaB was successfully degraded by ClpYQ protease activity using ClpYQ in vitro and in vivo degradation assay. These findings demonstrated the YbaB is a novel substrate of ClpYQ protease. This work also successfully demonstrated that with the use of recognition element of a protease can successfully screen its substrates by indirect proteome chip screening assay.

頁(從 - 到)113-120
期刊Molecular and Cellular Proteomics
出版狀態Published - 2017 1月

All Science Journal Classification (ASJC) codes

  • 分析化學
  • 生物化學
  • 分子生物學


深入研究「Escherichia coli proteome microarrays identified the substrates of clpyq protease」主題。共同形成了獨特的指紋。