Abstract
The shark troponin I gene (TnI) was found for the first time in this study to inhibit endothelial cell proliferation and angiogenesis. This shark TnI had 68.9% amino acid homology with human TnI, whereas the polypeptide from Lys91 to Leu123, which is thought to be the active site of TnI, had 78.8% homology with the corresponding fragment of human TnI. However, the polypeptide of shark had higher activity to inhibit the proliferation of HUVEC and tumor cell lines than that of human TnI. To investigate the anti-angiogenesis and anti-tumor effect of the shark TnI polypeptide, the DNA sequence of polypeptide (Lys91-Leu123) of white-spot catshark TnI(psTnI) was cloned and fused with the His-SUMO cDNA, followed by expression in Escherichia coli. After its purification by Ni2+ affinity chromatography, the fusion His-SUMO-psTnI protein was digested with the SUMO enzyme to release psTnI. The inhibitory ability of this recombinant shark TnI polypeptide for angiogenesis was confirmed by chicken embryo allantoic membrane (CAM) test and IHC analysis. It was also found by breast carcinoma xenograft study in Balb/c mice that this polypeptide could inhibit tumor growth in vivo.
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Acknowledgments
The authors would like to thank Dr. Huang Yadong to offer us the gene of SUMO. We are very grateful for the help of Dr. Su Zhijian on the purification of psTnI and for the revision of this article by Prof. Cai Dongqing and Prof. Wei Xing. This study was supported by 211 Grant of MOE of China.
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Xie, Q., Yao, S., Chen, X. et al. A polypeptide from shark troponin I can inhibit angiogenesis and tumor growth. Mol Biol Rep 39, 1493–1501 (2012). https://doi.org/10.1007/s11033-011-0887-y
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DOI: https://doi.org/10.1007/s11033-011-0887-y