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《China Biotechnology》 2009-08
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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-ping3CHEN Chang-you1 LIU Jin-song1 XU Ai-min1,4 WU Dong-hai1,2(1 Guangzhou Institute of Biomedicine and Health,Chinese Academy of Science,Guangzhou 510663,China)(2 Department of Life Science,University of Science and Technology of China,Hefei 230026,China)(3 Liver Disease Center of PLA,PLA 458 Hospital,Guangzhou 510602,China)(4 Department of Medicine,University of Hong Kong,Hong Kong 999077,China)  
APPL proteins,including APPL1 and APPL2,are important messengers between cell membrane and cell nucleus and are essential to cell proliferation.The PTB domain of human APPL2(hAPPL2)in Escherichia coli was expressed and purified.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.It shed light on structural interactions between the PTB domain of APPL2 and the binding proteins.
【Fund】: 国家“973”计划资助项目(2007CB914301 2006CB910202 2004CB720102)
【CateGory Index】: R341
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