Research Article Open Access

Expression, Purification and Activity Assay of Two New Recombinant Antagonists of Fibrinogen Receptor

Jianbo Yang1, Jia Yao1, Kun Yang1, Zichun Hua1 and Jie Yang1
  • 1 Nanjing University, China

Abstract

The gene sequence of Decorsin which is extracted from a kind of North American leeches was synthesized. Two recombinant proteins, Annexin V plus Decorsin (AnnV-D39) and Annexin V plus the carboxyl terminal 27 amino acid residues of Decorsin(AnnV-D27), were constructed. And a 10 amino acids linker peptide of GGGGSGGGGS was inserted between Annexin V and Decorsin in AnnV-D39. Using pET-28(a+) as an expressing vector, both two recombinant proteins were expressed in E. Coli BL21(DE3) with high efficiency as inclusion bodies. The expression products were purified by DEAE-Cellulose 52 and Sepharose CL-4B chromatography under denaturing condition. Platelet Aggregation Assay (PAA) shows that AnnV-D39 has good anti-platelet aggregation activity. However, AnnV-D27 shows no such activities in any PAA test. AnnV-D39 shows good anti-platelet aggregation activity as a new antagonist of fibrinogen receptor, while Annv-D27 needs re-modification.

American Journal of Biochemistry and Biotechnology
Volume 1 No. 2, 2005, 69-73

DOI: https://doi.org/10.3844/ajbbsp.2005.69.73

Submitted On: 20 April 2005 Published On: 30 June 2005

How to Cite: Yang, J., Yao, J., Yang, K. & Hua, Z. (2005). Expression, Purification and Activity Assay of Two New Recombinant Antagonists of Fibrinogen Receptor. American Journal of Biochemistry and Biotechnology, 1(2), 69-73. https://doi.org/10.3844/ajbbsp.2005.69.73

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Keywords

  • Antagonist of fibrinogen receptor
  • anti-platelet aggregation activity
  • annv-Decorsin fusion