Synthesis of a new zinc finger peptide; comparison of its `code' deduced and `CASTing' derived binding sites

• Published in: FEBS letters 1997-11, Vol.417 (1), p.71-74
• Main Authors: Corbi, Nicoletta, Perez, Marie, Maione, Rossella, Passananti, Claudio
• Format: Article
• Language: English
• Subjects: Amino Acid Sequence; Base Sequence; Binding Sites; Casting selection; Dissociation constant; DNA - metabolism; DNA-Binding Proteins - chemical synthesis; DNA-Binding Proteins - chemistry; DNA-Binding Proteins - genetics; DNA-Binding Proteins - metabolism; Molecular Sequence Data; Peptides - chemical synthesis; Peptides - chemistry; Peptides - genetics; Peptides - metabolism; Protein design; Protein Engineering; Recognition code; Zinc finger-DNA interaction; Zinc Fingers
• ISSN: 0014-5793; 1873-3468
• DOI: 10.1016/S0014-5793(97)01257-X

Using two synthetic oligonucleotides, we have constructed a new gene containing three zinc finger motifs of the Cys 2-His 2 type. We named this artificial gene `Mago'. The Mago nucleotide triplets encoding the amino acid positions, described to be crucial for DNA binding specificity, have been chosen on the basis of the proposed recognition `code' that relates the zinc finger's primary structure to the DNA binding target. Here we demonstrate that Mago protein specifically binds the `code' DNA target, with a dissociation constant ( K d) comparable to the K d of the well known Zif268 protein with its binding site. Moreover, we show that the deduced Mago `code' and the `experimental' selected DNA binding sites are almost identical, differing only in two nucleotides at the side positions.