Citation: Xiao Ping Wu, Xiao Kun Li. Effect of charge at an amino acid of basic fibroblast growth factor on its mitogenic activity[J]. Chinese Chemical Letters, ;2010, 21(4): 468-471. doi: 10.1016/j.cclet.2009.12.004 shu

Effect of charge at an amino acid of basic fibroblast growth factor on its mitogenic activity

Xiao Ping Wu ^a, ,
Xiao Kun Li ^b

a Institute of Tissue Transplantation and Immunology, Jinan University, Guangzhou 510632, China;
b National Engineering Research Center for Gene Medicine, Jinan University, Guangzhou 510632, China

Received Date: 26 August 2009

Fund Project: The work was supported by grants from the Natural Science Foundation of China (No. 30973671), the Natural Science Foundation of Guangdong Province of China (No. 9151064001000031), and Guangdong Provincial Key Discipline in Biochemistry and Molecular biology.

The amino acid at the 119th position of human basic fibroblast growth factor(hbFGF),lysine(K119),is a critical component for its mitogenic activity.However,little is known about the effects of the characteristics of this residue including charge on the mitogenic activity of hbFGE Herein,this basic residue was replaced with neutral glutamine residue and acidic glutamic acid residue to construct mutants hbFGFK^119Q and hbFGFK^119E,respectively.The mutants were produced by BL21(DE3)/pET3c expression system and purified to homogeneity by ionic exchange and heparin affinity chromatography from the supernatant of bacteria lysate.The mitogenic activity analysis showed that neutralization of charge at the 119th residue diminished the mitogenic activity of hbFGE whereas change of positive charge to negative charge at this residue had no significant effect on its mitogenic activity.Further MAP kinase activation assay revealed that the influence of different charge at the 119th position on the mitogenic activity of hbFGF was related to the signal molecular activation in MAPK pathway.It was deduced that the charge,either positive or negative,at the 119th position of hbFGF is crucial for its full mitogenic activity.
- Human basic fibroblast growth factor,
- Amino acid charge,
- Expression,
- Mitogenic activity,
- Erk1/2 activation
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