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[Hepatocellular carcinoma associated antigen HCA520, a novel Ca2+-binding protein]
Mei-xiang Yang1, Ke-jun Han, Xun Qu
1Basic Research Institute of Clinic Medicine, Qilu Hospital, Shandong University, Jinan 250012, China. ymxwjq@163.com
Aim:
To analyze the binding activity of the hepatocellular carcinoma associated antigen HCA520 to Ca2+.
Methods:
The HCA520 gene was gained by PCR. Then, it was cloned into the prokaryotic expression vector pGEX-4T-3. The GST-HCA520 fusion gene was induced to express in E. coli and the expressed product was purified via GST-agarose affinity resin. The fusion protein was confirmed by Western blot analysis. The Ca2+ binding capability of the fusion protein was analyzed by dot blot.
Results:
The GST-HCA520 fusion gene was constructed and the protein was successfully expressed and purified, which was identified by DNA sequencing and Western blot respectively. The fusion protein could bind to Ca2+ depend on dosages.
Conclusion:
HCA520, a novel hepatocellular carcinoma associated antigen, is a novel Ca2+ binding protein.
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