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Monitoring the cellular dynamic distribution of mammalian Ste20‐like protein kinase 3 by peptide aptamer from phage display
Author(s) -
Chiu YiRong,
Yuan ChiunJye
Publication year - 2010
Publication title -
the faseb journal
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.709
H-Index - 277
eISSN - 1530-6860
pISSN - 0892-6638
DOI - 10.1096/fasebj.24.1_supplement.466.1
Subject(s) - phage display , fusion protein , microbiology and biotechnology , biology , peptide , subcellular localization , cytoplasm , biochemistry , gene , recombinant dna
Mammalian Ste20‐like protein kinase 3 (Mst3), a member of the GC‐III subfamily protein kinases, is found to regulate various cellular affairs in response to environmental cues. The localization of endogenous Mst3 is in cytoplasm and may translocate to nucleus upon stimulation. We are interested in screening Mst3‐binding peptide for future drug development. Two phage display libraries, hepta‐peptide and dodeca‐peptide phage libraries had been subjected to an in vitro selection process for Mst3‐binding ligand identification. Following six biopanning process of Mst3‐binding selection and DNA sequencing of selected phages, five Mst3‐binding phages were selected. The five types of Mst3‐binding phages had been confirmed that all could bind with Mst3 but not Mst4 by ELISA assay and Western blotting. The peptides were constructed into EGFP vector to form EGFP fusion proteins in vivo . The peptide‐EGFP fusion proteins will be used to real time monitor the subcellular redistribution of endogenous Mst3 in response to various environmental stimuli.