High-level expression of a recombinant fragment of human fibronectin containing the Cell I–Hep II–IIICS71 domain in Escherichia coli as a soluble protein (original) (raw)

Abstract

Fibronectin (FN) is a major matrix protein that is involved in multiple processes. Its Cell I–Hep II domain is potentially useful in tumor therapy. Here, a recombinant fragment of FN with the Cell I–Hep II-IIICS71 domain, CH/71, was expressed in Escherichia coli. The CH/71 fusion protein consists of Cell I–Hep II domain and 19th to 89th amino acids of IIICS domain of FN. The expression level of CH/71 in E. coli was very high after induction with IPTG. Furthermore, CH/71 protein was largely found in the soluble fraction. It was readily purified by one-step heparin–agarose affinity chromatograph. The ability of CH/71 binding cells was about 8-fold of that of Cell I–Hep II domain FN.

Access this article

Log in via an institution

Subscribe and save

• Starting from 10 chapters or articles per month
• Access and download chapters and articles from more than 300k books and 2,500 journals
• Cancel anytime View plans

Buy Now

Price excludes VAT (USA)
Tax calculation will be finalised during checkout.

Instant access to the full article PDF.

Similar content being viewed by others

References

• Abe Y, Bui-Thanh NA, Ballantyne CM, Burns AR (2005) Extra domain A and type III connecting segment of fibronectinin assembly and cleavage. Biochem Biophys Res Commun 338:1640–1647
  Article PubMed CAS Google Scholar
• Chung CP, Jang JH (2005) Design and expression of oligomeric fibronectin fusion protein: a strategy for enhancing cell adhesion activity. Biotechnol Lett 27:811–816
  Article PubMed CAS Google Scholar
• Feng Z, Zhang G, Li D, Zhang H (1996) Construction of expressing plasmids of recombinant FN polypeptides with bifunctional-domain and the characterization of the products expressed in E. coli. J Tongji Med Univ 16:70–74, 86
  Google Scholar
• Hynes RO (1990) Fibronectins. Springer-Verlag, New York
  Google Scholar
• Hynes RO (2002) Integrins: bidirectional, allosteric signaling machines. Cell 110:673–687
  Article PubMed CAS Google Scholar
• Kimizuka F, Taguchi Y, Ohdate Y, Kawase Y, Shimojo T, Hashino K, Kato I, Sekiguchi K, Titani K (1991) Production and characterization of functional domains of human fibronectin expressed in Escherichia coli. J Biochem 110:284–291
  PubMed CAS Google Scholar
• Labat-Robert J (2002) Fibronectin in malignancy. Semin Cancer Biol 12:187–195
  Article PubMed CAS Google Scholar
• Liu X, Collodi P (2002) Novel form of fibronectin from zebrafish mediates infectious hematopoietic necrosis virus infection. J Virol 76:492–498
  Article PubMed CAS Google Scholar
• Mao Y, Schwarzbauer JE (2005) Fibronectin fibrillogenesis: a cell-mediated matrix assembly process. Matrix Biol 24:389–399
  Article PubMed CAS Google Scholar
• Saiki I, Makabe T, Yoneda J (1991) Inhibitory effect of fibronectin and its recombinant polypeptides on the adhesion of metastatic melanoma cells to laminin. Jpn J Cancer Res 82:1112–1119
  PubMed CAS Google Scholar
• Sakai T, Johnson KJ, Murozono M, Sakai K, Magnuson MA, Wieloch T, Cronberg T, Isshiki A, Erickson HP, Fassler R (2001) Plasma fibronectin supports neuronal survival and reduces brain injury following transient focal cerebral ischemia but is not essential for skin-wound healing and hemostasis. Nat Med 7:324–330
  Article PubMed CAS Google Scholar
• Sambrook J, Fritsch EF, Maniatis T (2001) Molecular cloning: a laboratory manual, 3rd ed. Cold Spring Harbor Laboratory Press, New York
  Google Scholar
• Schein CH (1993) Solubility and secretability. Curr Opin Biotechnol 4:456–461
  Article PubMed CAS Google Scholar
• Schwarzbauer JE (1991) Fibronectin: from gene to protein. Curr Opin Cell Biol 3:786–791
  Article PubMed CAS Google Scholar
• Yi M, Ruoslahti E (2001) A fibronectin fragment inhibits tumor growth, angiogenesis, and metastasis. Proc Natl Acad Sci USA 98:620–624
  Article PubMed CAS Google Scholar
• Yoneda J, Saiki I, Igarashi Y, Kobayashi H, Fujii H, Ishizaki Y, Kimizuka F, Kato I, Azuma I (1995) Role of the heparin-binding domain of chimeric peptides derived from fibronectin in cell spreading and motility. Exp Cell Res 217:169–179
  Article PubMed CAS Google Scholar
• Zhang G, Feng Z, Li D, Zhang H (2000) Comparison of effects of CH50 on macrophage activation and its anti-tumor activity with those of lipopolysaccharides. Acta Pharmacol Sin 21:567–570
  PubMed CAS Google Scholar
• Zhang G, Yang Y, Huang B, Xiao H, Li D, Feng Z (2003) Experimental study on therapeutic effect of in vivo expression of Cell I–Hep II recombinant polypeptide of fibronectin on murine H22 hepatocellular carcinoma. World J Gastroenterol 9:1940–1945
  PubMed CAS Google Scholar

Download references

Acknowledgements

This work was supported by grants from National Natural Science Foundation of China (39370783) and the Medical Science Foundation of Guangdong Province, China (No. A2003484).

Author information

Authors and Affiliations

1. Allergy and Inflammation Research Institute, Shantou University Medical College, Shantou, 515041, P.R. China
   Mingcai Li
2. Department of Biochemistry and Molecular Biology, Tongji Medical College, Huazhong University of Science and Technology, Wuhan, 430030, P.R. China
   Mingcai Li, Zuohua Feng, Guimei Zhang & Dong Li

Authors

1. Mingcai Li
2. Zuohua Feng
3. Guimei Zhang
4. Dong Li

Corresponding author

Correspondence toMingcai Li.

Rights and permissions

About this article

Cite this article

Li, M., Feng, Z., Zhang, G. et al. High-level expression of a recombinant fragment of human fibronectin containing the Cell I–Hep II–IIICS71 domain in Escherichia coli as a soluble protein.Biotechnol Lett 28, 1141–1146 (2006). https://doi.org/10.1007/s10529-006-9066-y

Download citation

• Received: 07 February 2006
• Accepted: 31 March 2006
• Published: 23 June 2006
• Issue date: July 2006
• DOI: https://doi.org/10.1007/s10529-006-9066-y

Key words