Selection and identification of human Gonadotropin‐releasing hormone promoter binding peptides by phage display‐CEMSA

Abstract

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.

Recent developments in CE and CEC of peptides

“…CAE mobility shift assay has been found to be an efficient and sensitive method also for both qualitative and quantitative studies of the interactions between the peptides and oligo-and polynucleotides, it has been employed, e.g., for selection and identification of human GnRH promoter binding peptides using -813/-1081 region of hGnRH promoter and a phage-displayed peptide library [253] and for selection of aptamers with high affinity to neuropeptide Y from random-sequence nucleic acid libraries [254].…”

Selection of staphylococcal enterotoxin B (SEB)-binding peptide using phage display technology

“…Phage display technology has been used for selection and production of molecules that have antibody characteristics. Recombinant antibodies have been selected against various analytes like staphylococcal enterotoxin B [6], spores of Bacillus anthracis [7], Art v1 glycoprotein [8], rotavirus NSP4 enterotoxin [9], coronavirus [10], Clostridium botulinum neurotoxin serotype A [11], Salmonella typhimurium [12], and human Gonadotropin-releasing hormone promoter [13] by phage display technology. It also has been used to design high affinity super antigens for immunochemical application and immunotherapy [14] and to choose specific molecules to inorganic materials [2].…”