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Expression of the Pro-Domain?Deleted Active Form of Caspase-6 in Escherichia coli
Expression of the Pro-Domain?Deleted Active Form of Caspase-6 in Escherichia coli
Journal of Microbiology and Biotechnology. 2014. May, 24(5): 719-723
Copyright © 2014, The Korean Society For Microbiology And Biotechnology
  • Received : December 13, 2013
  • Accepted : February 26, 2014
  • Published : May 30, 2014
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About the Authors
Phil Young Lee
Medical Proteomics Research Center, KRIBB, Yusung, Daejeon 305-806, Republic of Korea
Jin Hwa Cho
Medical Proteomics Research Center, KRIBB, Yusung, Daejeon 305-806, Republic of Korea
Seung Wook Chi
Medical Proteomics Research Center, KRIBB, Yusung, Daejeon 305-806, Republic of Korea
Kwang-Hee Bae
Cell Function Regulation Research Center, KRIBB, Yusung, Daejeon 305-806, Republic of Korea
Sayeon Cho
College of Pharmacy, Chung-Ang University, Seoul 156-756, Republic of Korea
Byoung Chul Park
Medical Proteomics Research Center, KRIBB, Yusung, Daejeon 305-806, Republic of Korea
Jeong-Hoon Kim
Medical Proteomics Research Center, KRIBB, Yusung, Daejeon 305-806, Republic of Korea
sgpark@kribb.re.kr
Sung Goo Park
Medical Proteomics Research Center, KRIBB, Yusung, Daejeon 305-806, Republic of Korea
sgpark@kribb.re.kr

Abstract
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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Acknowledgements
This work was supported by KRIBB and research grants from the National Research Foundation of Korea (NRF-2011-0028172).
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