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| Expression,Purification of PTB Domain of Human APPL2 and Screen for Its Peptide Ligands |
LIANG Ning1,2 SHI Zhu-liang1,2 WANG Dong-ye1 WEI Guo-wei1 KONG Xiang-ping3
CHEN Chang-you1 |LIU Jin-song1 XU Ai-min1,4 WU Dong-hai1,2 |
1 Guangzhou Institute of Biomedicine and Health,Chinese Academy of Science,Guangzhou510663,China
2 Department of Life Science,University of Science and Technology of China,Hefei230026,China
3 Liver Disease Center of PLA,PLA 458 Hospital,Guangzhou510602,China
4 Department of Medicine,University of Hong Kong,Hong Kong999077,China |
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Abstract APPL proteins, including APPL1 and APPL2, are important messengers between cell membrane and cell nucleus and are essential to cell proliferation. In this study, we expressed and purified the PTB domain of human APPL2 (hAPPL2) in Escherichia coli. Phage display technology was then used to screen for the interacting peptides with this domain. After 3 rounds of panning, 48 single phage plaques were randomly picked and ELISA analysis was performed. Two 7-mer peptides, ERLPFFY and YLTSPKH were found to bind to the PTB domain of hAPPL2 in a specific manner. Both wild type and mutant forms of P3-GST fusion protein, with each amino acid substituted by alanine, were prepared and evaluated for binding capacity with hAPPL2 PTB by ELISA. Results showed that each residue was indispensible in the binding between this peptide to hAPPL2 PTB domain. This study shed light on structural interactions between the PTB domain of APPL2 and the binding proteins.
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Received: 04 March 2009
Published: 28 July 2009
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Corresponding Authors:
Ning Liang
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