Expression and purification of a biologically active basic fibroblast growth factor fusion protein.
Basic fibroblast growth factor (bFGF) is a potent mitogen of many cell types and plays an important role in angiogenesis. To help identify proteins that bind to bFGF and mediate its intracellular transport and signaling, we overexpressed and purified a bFGF fusion protein in Escherichia coli. The fusion protein consists of bFGF fused to the C-terminus of glutathione S-transferase (GST). The GST-bFGF fusion protein was purified using SP-Sepharose and glutathione-Sepharose affinity chromatography. The ability of the purified GST-bFGF to stimulate the growth of human umbilical vein endothelial cells (HUVECs) was equivalent to that of purified recombinant 18 kDa bFGF.
Sheng
Zhi
Z
Department of Anatomy and Cell Biology, State University of New York, Downstate Medical Center, Box 5, 450 Clarkson Avenue, Brooklyn, NY 11203, USA.
Chang
Shin-Bey
SB
Chirico
William J
WJ
D015153:Blotting, Western; D002455:Cell Division; D002478:Cells, Cultured; D002847:Chromatography, Agarose; D018076:DNA, Complementary / Q000378:metabolism; D004305:Dose-Response Relationship, Drug; D004586:Electrophoresis; D004730:Endothelium, Vascular / Q000166:cytology; D004926:Escherichia coli / Q000378:metabolism; D016222:Fibroblast Growth Factor 2 / Q000737:chemistry* / Q000378:metabolism; D005982:Glutathione Transferase / Q000378:metabolism; D006801:Humans; D010957:Plasmids / Q000378:metabolism; D011485:Protein Binding; D017434:Protein Structure, Tertiary; D011993:Recombinant Fusion Proteins / Q000737:chemistry* / Q000378:metabolism; D011994:Recombinant Proteins / Q000378:metabolism; D014471:Umbilical Veins / Q000166:cytology
D018076:DNA, Complementary; D011993:Recombinant Fusion Proteins; D011994:Recombinant Proteins; D016222:Fibroblast Growth Factor 2; D005982:Glutathione Transferase