Production of Bioactive Rockfish (Sebastes schlegeli) Myostatin-1 Prodomain in an Escherichia coli System

被引：4
作者：
Lee, Sang Beum [1 ]
Cho, Mi-Jin [1 ]
Kim, Jeong Hwan [1 ]
Kim, Yong Soo [2 ]
Jin, Hyung-Joo [1 ]
机构：
[1] Gangneung Wonju Natl Univ, Div Marine Mol Biotechnol, Fac Marine Biosci & Technol, Kangnung 210702, South Korea
[2] Univ Hawaii, Dept Human Nutr Food & Anim Sci, Honolulu, HI 96822 USA
来源：
PROTEIN JOURNAL | 2011年 / 30卷 / 01期
关键词：
Myostatin; GDF-8; Prodomain; Rockfish; Sebastes schlegeli; SKELETAL-MUSCLE MASS; PROPEPTIDE GENE; GROWTH; PROTEINS; MUTATION; FOLLISTATIN; EXPRESSION; DELIVERY; VECTORS; CATTLE;
D O I：
10.1007/s10930-010-9301-1
中图分类号：
Q5 [生物化学]; Q7 [分子生物学];
学科分类号：
071010 ; 081704 ;
摘要：
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.
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页码：52 / 58
页数：7
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