Construction of expressing plasmids of recombinant FN polypeptides with bifunctional-domain and the characterization of the products expressed inE. coli

Abstract

Two expressing plasmids have been constructed and used to express two bifurictional-domain recombinant polypeptides of human fibronectin (FN) inE. coli. One was CH50 (Pro1239-Ser1515 of FN linked with Ala1690-Thr1960 of FN through Met) and the other was CH56 (Pro1239-Thr1960 of FN). Both of two polypeptides were capable of binding heparin and were purified by heparin-agarose affinity chromatography. The purified products were capable of binding cells. The production of CH50 and CH56 polypeptides provided a fundamental basis for further study of the anti-metastatic function of recombinant fibronectin polypeptides.

Keywords

fibronectin / recombinant polypeptide / metastasis

References

[1] KimizukaF, TaguchiY, OhaateY, et al.. Production and characterization of functional domains of human fibronection expressed in Escherichia coli. J Biochem, 1991, 110: 284-284

[2] SaikiI, MakabeT, YonedaJ, et al.. Inhibitory effect of fibronection and its recombinant polypeptides on the adhesion of metastatic melanoma cells to laminin. Jpn J Cancer Res, 1991, 82: 1-112

[3] MatsumotoY, SaikiY, MakabeT, et al.. Inhibitory effect of antimetastatic fusion polypeptide of human fibronectin on tumor cell adhesion to extracellular matrices. Jpn J Cancer Res, 1991, 82: 1-130

[4] YonedaJ, SaikiI, KobayashiH, et al.. Inhibitory effect of recombinant fibronectin polypeptides on the adhesion ofliver-metastatic lymphoma cells to hepatic sinusoidal endothelial cells and tumor invasion. Jpn J Cancer Res, 1994, 85: 723-723

[5] SaikiI, MurataJ, MakabeT, et al.. Inhibition of lung metastasis by synthetic and recombinant fragments of human fibronectin witn functional domains. Jpn J Cancer Res, 1990, 81: 1-003

[6] SaikiI, MatsumotoY, MurataJ, et al.. Recombinant fusion polypeptide with Cell- and heparin-binding domains of fibronectin inhibits liver metastasis of L5178Y-ML25 lymphoma cells. Jpn J Cancer Res, 1991, 82: 1-120

[7] KornblihttA, UmezawaK, Vibe-PedersenK, et al.. Primary structure of human fibronectin: differential splicing may generate at least 10 polypeptides from a single gene. The EMBO J, 1985, 4(7): 1-755

[8] SambrookJ, FritchEF, ManiatisT. Molecular Cloning, a laboratory manu, 19892nd edWuhanCold Spring Harbor Laboratory Press1.21-1.87

[9] HynesR O. Fibronectins, 1990WuhanHuazhong University of Science and Technology New York Inc113-115

[10] BeezholdDH, PersoniusC. Fibronectin fragments stimulate tumor necrosis factor secretion by human monocytes. J Leukoc Biol, 1992, 51(1): 59-59