[Hepatocellular carcinoma associated antigen HCA520, a novel Ca2+-binding protein].
OBJECTIVE
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.


CONCLUSIONS
HCA520, a novel hepatocellular carcinoma associated antigen, is a novel Ca2+ binding protein.
Yang
Mei-xiang
MX
Basic Research Institute of Clinic Medicine, Qilu Hospital, Shandong University, Jinan 250012, China. ymxwjq@163.com
Han
Ke-jun
KJ
Qu
Xun
X
Ma
Dao-xin
DX
Chen
Wei-feng
WF
D002118:Calcium / Q000378:metabolism*; D002135:Calcium-Binding Proteins / Q000235:genetics / Q000302:isolation & purification / Q000378:metabolism*; D006528:Carcinoma, Hepatocellular / Q000737:chemistry; D003001:Cloning, Molecular; D004926:Escherichia coli / Q000378:metabolism*; D005822:Genetic Vectors; D005982:Glutathione Transferase / Q000235:genetics / Q000378:metabolism; D006801:Humans; D008113:Liver Neoplasms / Q000737:chemistry; D009363:Neoplasm Proteins / Q000235:genetics / Q000302:isolation & purification / Q000378:metabolism*; D009857:Oncogenes; D011485:Protein Binding; D011993:Recombinant Fusion Proteins / Q000235:genetics / Q000378:metabolism
C469743:CHP1 protein, human; D002135:Calcium-Binding Proteins; D009363:Neoplasm Proteins; D011993:Recombinant Fusion Proteins; D005982:Glutathione Transferase; D002118:Calcium