title: Production of recombinant human apoptosis signal-regulating kinase 1 (ASK1) in Escherichia coli.
abstract: Apoptosis signal-regulating kinase 1 (ASK1) is a mediator of the MAPK signaling cascade, which regulates different cellular processes including apoptosis, cell survival, and differentiation. The increased activity of ASK1 is associated with a number of human diseases and this protein kinase is considered as promising therapeutic target. In the present study, the kinase domain of human ASK1 was expressed in Escherichia coli (E. coli) in soluble form. The expression level of ASK1 was around 0.3-0.47 g per 1 L after using auto-induction protocol or IPTG induction. A one-step on column method for the efficient purification of recombinant ASK1 was performed. Our approach yields sufficient amount of recombinant ASK1, which can be used for inhibitor screening assays and different crystallographic studies.
authors.affiliation: Institute of Molecular Biology and Genetics, NAS of Ukraine, 150 Zabolotnogo St., 03680, Kyiv, Ukraine. Electronic address: galina.volinetc@gmail.com.
authors.forename: Galyna P
authors.initials: GP
authors.lastname: Volynets
authors.affiliation: Institute of Molecular Biology and Genetics, NAS of Ukraine, 150 Zabolotnogo St., 03680, Kyiv, Ukraine; The State Institute of Genetic and Regenerative Medicine, NAMS of Ukraine, Ukraine.
authors.forename: Oksana B
authors.initials: OB
authors.lastname: Gorbatiuk
authors.affiliation: Institute of Molecular Biology and Genetics, NAS of Ukraine, 150 Zabolotnogo St., 03680, Kyiv, Ukraine.
authors.forename: Oleksandr P
authors.initials: OP
authors.lastname: Kukharenko
authors.affiliation: Institute of Molecular Biology and Genetics, NAS of Ukraine, 150 Zabolotnogo St., 03680, Kyiv, Ukraine.
authors.forename: Mariya O
authors.initials: MO
authors.lastname: Usenko
authors.affiliation: Institute of Molecular Biology and Genetics, NAS of Ukraine, 150 Zabolotnogo St., 03680, Kyiv, Ukraine.
authors.forename: Sergiy M
authors.initials: SM
authors.lastname: Yarmoluk
mesh_terms: D004926:Escherichia coli / Q000235:genetics / Q000378:metabolism; D015870:Gene Expression; D006801:Humans; D048848:MAP Kinase Kinase Kinase 5 / Q000096:biosynthesis / Q000737:chemistry / Q000235:genetics / Q000302:isolation & purification; D000072417:Protein Domains; D011994:Recombinant Proteins / Q000096:biosynthesis / Q000737:chemistry / Q000235:genetics / Q000302:isolation & purification
chemical_list: D011994:Recombinant Proteins; D048848:MAP Kinase Kinase Kinase 5; C482858:MAP3K5 protein, human
keywords: ASK1; Apoptosis signal-regulating kinase 1; Auto-induction protocol; IPTG induction; Recombinant protein