Efficient expression of soluble human FGF-21 and its glucose regulation activity.
- Author:
Gui-ping REN
1
;
Yu-ting HOU
;
Yuan-yuan JIANG
;
Jin-nan LI
;
Wei ZHANG
;
Liu DI
;
De-shan LI
Author Information
1. College of Life Science of Northeast Agricultural University, Provincial Academy of Agricultural Sciences of Heilongjiang Postdoctoral Station, Northeast Forestry University Postdoctoral Station, Harbin 150030, China.
- Publication Type:Journal Article
- MeSH:
3T3-L1 Cells;
Adipocytes;
metabolism;
Animals;
DNA, Complementary;
genetics;
Escherichia coli;
genetics;
metabolism;
Fibroblast Growth Factors;
genetics;
metabolism;
pharmacology;
Glucose;
metabolism;
Humans;
Mice;
Plasmids;
Recombinant Fusion Proteins;
genetics;
metabolism;
pharmacology;
Small Ubiquitin-Related Modifier Proteins;
genetics;
metabolism;
Solubility
- From:
Acta Pharmaceutica Sinica
2009;44(5):548-552
- CountryChina
- Language:Chinese
-
Abstract:
The cDNA of human FGF-21 was subcloned into the pSUMO expression vector and the fusion protein was induced to express in Escherichia coli Rosetta (DE3). The recombinant hFGF-21 was expressed in soluble form in the pSUMO expression system. The recombinant fusion protein was purified by Ni-NTA column. The purified recombinant protein was dialyzed against PBS for re-nature. To obtain pure and active recombinant protein, the fusion protein was subjected to cleavage with SUMO protease I. To examine glucose regulation activity of hFGF-21, 3T3-L1 pre-adipocytes were differentiated into adipocytes, glucose up-take activity of hFGF-21 was examined by glucose oxidase and peroxidase (GOD-POD) assay. Compared with no stimulation control, the recombinant hFGF-21 treatment led to a significant increase in glucose consumption of adipocytes and a significant decrease in concentration of glucose in the medium (P < 0.05, P < 0.001).
