title: Expression and Efficient One-Step Chromatographic Purification of a Soluble Antagonist for Human Leukemia Inhibitory Factor Receptor in Escherichia coli.
abstract: Leukemia inhibitory factor (LIF) is a member of the IL-6 cytokine family, having pleiotropic actions such as maintaining stem cell pluripotency and enabling blastocyst implantation. Because the action of LIF is mediated by a ligand-receptor interaction with the LIF receptor (LIF-R), an antagonist for LIF-R has been developed to inhibit LIF-induced signaling. In this study, we present a novel method for the production and purification of an antagonist to human LIF-R (hLA). His-tagged hLA was expressed in E. coli, and simple purification methods without any endopeptidase cleavage were designed. In addition, we determined the optimal temperature conditions for enhancing the production of soluble hLA. Finally, the bioactivity of His-tagged hLA was examined using STAT3 phosphorylation and receptivity of human endometrial ECC-1 cells. Our strategy provides a rapid and efficient method to produce biologically active recombinant hLA.
authors: Department of Korean Medical Science, School of Korean Medicine and Korean Medicine Research Center for Healthy Aging, Pusan National University, Yangsan 626-870, Republic of Korea.
authors: Eun-Yeong
authors: EY
authors: Kim
authors: Department of Korean Medical Science, School of Korean Medicine and Korean Medicine Research Center for Healthy Aging, Pusan National University, Yangsan 626-870, Republic of Korea.
authors: Hee-Jung
authors: HJ
authors: Choi
authors: Department of Korean Medical Science, School of Korean Medicine and Korean Medicine Research Center for Healthy Aging, Pusan National University, Yangsan 626-870, Republic of Korea.
authors: Tae-Wook
authors: TW
authors: Chung
authors: Department of Molecular Biology, College of Natural Sciences, Pusan National University, Busan 609-735, Republic of Korea.
authors: Se Bok
authors: SB
authors: Jang
authors: 2nd Division of Clinical Medicine, School of Korean Medicine, Pusan National University and Pediatric, Korean Medicine Hospital, Pusan National University Hospital, Yangsan 626-870, Republic of Korea.
authors: Kibong
authors: K
authors: Kim
authors: Department of Korean Medical Science, School of Korean Medicine and Korean Medicine Research Center for Healthy Aging, Pusan National University, Yangsan 626-870, Republic of Korea.
authors: Ki-Tae
authors: KT
authors: Ha
mesh_terms: D002460:Cell Line; D002845:Chromatography / Q000379:methods*; D004926:Escherichia coli / Q000235:genetics* / Q000378:metabolism; D015870:Gene Expression; D006801:Humans; D053606:Receptors, OSM-LIF / Q000037:antagonists & inhibitors*; D011994:Recombinant Proteins / Q000096:biosynthesis* / Q000302:isolation & purification* / Q000378:metabolism
chemical_list: D053606:Receptors, OSM-LIF; D011994:Recombinant Proteins
keywords: Antagonist; Endometrial cells; Escherichia coli; His-tagged; LIF-05; Leukemia inhibitory factor