Selection and identification of human gonadotropin-releasing hormone promoter binding peptides by phage display-CEMSA.
Specific interactions between transcription factors and cis-acting DNA sequences form the molecular basis of gene expression regulation. Here, we applied phage display technology to DNA-protein interaction studies. A phage-displayed peptide library was used to select Gonadotropin-releasing hormone promoter (GP) binding peptides. After four sequential rounds of biopanning on GP-conjugated magnetic beads, phage clones encoding GQPTPRNAGLPL (B6), SRLNVEPLTTYS (B3), and TTLHWASLTTGR (B11) were enriched. Phages bearing these peptides showed specific binding to GP in solution by capillary electrophoresis mobility shift assay (CEMSA). In addition, some human transcription factors were speculated as the potential transcription factors or co-activators of GnRH gene by bioinformatic analysis. These results suggest that phage display-CEMSA methodology should be a powerful tool to screen and identify site-specific DNA-binding peptides.
Xiao
Yichuan
Y
State Key Laboratory of Bioreactor Engineering, East China University of Science and Technology, 130 Meilong Road, Shanghai 200237, People's Republic of China.
Zhou
Yuxun
Y
Wang
Jianhui
J
Yu
Minghui
M
Wang
Gang
G
Jin
Jian
J
Xiao
Junhua
J
D000595:Amino Acid Sequence; D019921:Biotinylation; D002999:Clone Cells; D019295:Computational Biology; D019075:Electrophoresis, Capillary; D024202:Electrophoretic Mobility Shift Assay / Q000379:methods*; D004797:Enzyme-Linked Immunosorbent Assay; D007987:Gonadotropin-Releasing Hormone / Q000235:genetics*; D006801:Humans; D008969:Molecular Sequence Data; D019151:Peptide Library; D010455:Peptides / Q000032:analysis / Q000737:chemistry / Q000302:isolation & purification* / Q000378:metabolism*; D011401:Promoter Regions, Genetic / Q000235:genetics*; D011485:Protein Binding
D019151:Peptide Library; D010455:Peptides; D007987:Gonadotropin-Releasing Hormone