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| Location of EGFP Tag Affects the Binding Activity of Peptide Fusions |
| WANG Hai, SHI Bi-zhi, YANG Sheng-li, LI Zong-hai, LUO Xiao-ying |
| State Key Laboratory of Oncogenes and Related Genes, Renji Hospital, Shanghai Jiaotong University School of Medicine, Shanghai Cancer Institute, Shanghai 200032, China |
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Abstract Purpose: To investigate whether the location between targeting peptide and tag protein (eGFP) would influence the binding capacity of fusing protein to cell. Methods: To get Ge11 and LyP1 fusing eGFP recombination protein by expressing system in E coli. respectively, and then put the recombination protein into the medium of serum starving SMMC-7721 liver cancer cell line, after treating 3 hours, to detect the binding capacity by Fluorescence microscopy. Results: when eGFP at the Carboxyl terminal of targeting peptides, the recombination protein could bind to the cell; But when eGFP at the Nitro terminal, the recombination protein loss the binding capacity. On the basis of these researches, the other 3 targeting peptides were detected. Conclusion: These results indicate that an inappropriate fusion strategy would lead to decreased or even totally loss of function of targeting peptides. Thus, it should be cautious when used EGFP as a tracer to study the targeting activities of peptides.
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Received: 15 February 2012
Published: 25 June 2012
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