Expression, Purification and Activity Assay of Two New Recombinant Antagonists of Fibrinogen Receptor
Jianbo Yang, Jia Yao, Kun Yang, Zichun Hua and Jie Yang
DOI : 10.3844/ajbbsp.2005.69.73
American Journal of Biochemistry and Biotechnology
Volume 1, Issue 2
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.
© 2005 Jianbo Yang, Jia Yao, Kun Yang, Zichun Hua and Jie Yang. This is an open access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited.