title: Expression of the pro-domain-deleted active form of caspase-6 in Escherichia coli.
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.
authors.affiliation: Medical Proteomics Research Center, KRIBB, Yusung, Daejeon 305-806, Republic of Korea.
authors.forename: Phil Young
authors.initials: PY
authors.lastname: Lee
authors.forename: Jin Hwa
authors.initials: JH
authors.lastname: Cho
authors.forename: Seung Wook
authors.initials: SW
authors.lastname: Chi
authors.forename: Kwang-Hee
authors.initials: KH
authors.lastname: Bae
authors.forename: Sayeon
authors.initials: S
authors.lastname: Cho
authors.forename: Byoung Chul
authors.initials: BC
authors.lastname: Park
authors.forename: Jeong-Hoon
authors.initials: JH
authors.lastname: Kim
authors.forename: Sung Goo
authors.initials: SG
authors.lastname: Park
mesh_terms: D053178:Caspase 6 / Q000737:chemistry / Q000235:genetics* / Q000302:isolation & purification / Q000378:metabolism*; D004789:Enzyme Activation; D004926:Escherichia coli / Q000235:genetics* / Q000378:metabolism*; D015870:Gene Expression; D054730:Protein Interaction Domains and Motifs / Q000235:genetics*; D011994:Recombinant Proteins; D017384:Sequence Deletion
chemical_list: D011994:Recombinant Proteins; D053178:Caspase 6