title: Production of bioactive rockfish (Sebastes schlegeli) myostatin-1 prodomain in an Escherichia coli system.
abstract: Myostatin (MSTN) is a potent negative regulator of skeletal muscle growth in mammalian species, and its activity is inhibited by MSTN prodomain, the N-terminal part of proMSTN cleaved during post-translational MSTN processing. In fish, MSTN also appears to suppress fish muscle growth with its activity being inhibited by prodomain. The objective of this study was to produce bioactive MSTN-1 prodomain of rockfish (S. schlegeli), a commercial aquaculture species in East Asia, in E. coli using maltose binding protein (MBP) as a fusion partner. Rockfish MSTN-1 prodomain (sMSTN1pro) cDNA was cloned into the pMALc2x vector, and proteins (MBP-sMSTN1pro) were expressed in Rosetta-gami 2(DE3)pLysS cells by IPTG induction. The MBP-sMSTN1pro was expressed in soluble forms, and affinity purified using amylose resin. The affinity purified MBP-sMSTN1pro suppressed MSTN activity in vitro. The results suggest that MBP is probably a useful fusion partner in producing bioactive MSTN prodomains of various animal species in E. coli.
authors.affiliation: Division of Marine Molecular Biotechnology, Faculty of Marine Bioscience and Technology, Gangneung-Wonju National University, Gangneung 210-702, Korea.
authors.forename: Sang Beum
authors.initials: SB
authors.lastname: Lee
authors.forename: Mi-Jin
authors.initials: MJ
authors.lastname: Cho
authors.forename: Jeong Hwan
authors.initials: JH
authors.lastname: Kim
authors.forename: Yong Soo
authors.initials: YS
authors.lastname: Kim
authors.forename: Hyung-Joo
authors.initials: HJ
authors.lastname: Jin
mesh_terms: D000818:Animals; D004926:Escherichia coli / Q000235:genetics* / Q000378:metabolism; D029941:Fish Proteins / Q000037:antagonists & inhibitors / Q000096:biosynthesis* / Q000235:genetics* / Q000378:metabolism*; D058619:Maltose-Binding Proteins / Q000378:metabolism; D018482:Muscle, Skeletal / Q000378:metabolism*; D055435:Myostatin / Q000096:biosynthesis*
chemical_list: D029941:Fish Proteins; D058619:Maltose-Binding Proteins; D055435:Myostatin