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KMID : 0545120140240050719
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Journal of Microbiology and Biotechnology
2014 Volume.24 No. 5 p.719 ~ p.723
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Expression of the Pro-Domain?Deleted Active Form of Caspase-6 in Escherichia coli
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Lee Phil-Young
Cho Jin-Hwa
Chi Seung-Wook
Bae Kwang-Hee
Cho Sa-Yeon
Park Byoung-Chul
Kim Jeong-Hoon
Park Sung-Goo
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Abstract
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Caspases are a family of cysteine proteases that play an important role in the apoptotic pathway. Caspase-6 is an apoptosis effector that cleaves a variety of cellular substrates. The active form of the enzyme is required for use in research. However, it has been difficult to obtain sufficient quantities of active caspase-6 from Escherichia coli. In the present study, we constructed a caspase-6 with a 23-amino-acid deletion in the pro-domain. This engineered enzyme was expressed as a soluble protein in E. coli and was purified using affinity resin. In vitro enzyme assay and cleavage analysis revealed that the engineered active caspase-6 protein had characteristics similar to those of wild-type caspase-6. This novel method can be a valuable tool for obtaining active caspase-6 that can be used for screening caspase-6-specific substrates, which in turn can be used to elucidate the function of caspase-6 in apoptosis.
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KEYWORD
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caspase-6, active form, E. coli, enzyme assay
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