Prokaryotic Expression of PEX: a C-terminal Fragment of Matrix Metalloproteinase 2 and Its Effect on The Inhibition of Angiogenesis
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This work is supported by grants from the National “973” Program (G2000057002), National Natural Science Foundation of China (39880016), Natural Science Foundation of Beijing (7002009),Beijing High-Technology Key Laboratory of Molecular Cancer Biology、Bei

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    Abstract:

    To express chick MMP-2 C-terminal hemopexin-like fragment in E.coli and to explore whether it has the biological effect on the inhibition of angiogenesis, a 630 bp of MMP-2 C-terminal coding fragment (PEX) was cloned into prokaryotic expression vector pCal-n from primary cultured day 10 chicken embryo fibroblasts using RT-PCR. The relatively pure inclusion bodies of CBP/PEX fusion protein were denatured and renatured by guandine·HCl method from PEX/pCal-n plasmid transformed log phase E.coli BL21 (Des) pLys induced with IPTG. Growth curve of human umbilical vein endothelial cells (HUVEC) and chick embryo chorioallantoic membrane (CAM) assay were performed to examine the effects on the inhibition of angiogenesis. The prokaryotic expressed CBP/PEX fusion protein has the biological activity and can inhibit the growth of HUVEC in vitro and the angiogenesis on chick CAM in vivo. These results suggest that the expressed PEX is to be a novel potential therapeutic inhibitor of diseases associated with angiogenesis.

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LI Jin-Ping, ZHANG Guang-Mou, KE Yang, LIN Ben-Yao, ZHAO Wei, HU Ying, NING Tao, LIN Zhong-Xiang, ZHANG Zhi-Qian. Prokaryotic Expression of PEX: a C-terminal Fragment of Matrix Metalloproteinase 2 and Its Effect on The Inhibition of Angiogenesis[J]. Progress in Biochemistry and Biophysics,2002,29(1):120-123

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  • Received:April 25,2001
  • Revised:May 24,2001
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