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

FEBS Letters

Passananti, Claudio¹  Maione, Rossella²  Perez, Marie²  Corbi, Nicoletta² 

[1] Istituto di Tecnologie Biomediche, CNR, Via Morgagni 30/E, 00161 Rome, Italy;Dipartimento di Biotecnologie Cellulari ed Ematologia, Università di Roma La Sapienza, Viale Regina Elena 324, 00161 Rome, Italy

关键词: Zinc finger-DNA interaction;    Protein design;    Recognition code;    Casting selection;    Dissociation constant;   

DOI  :  10.1016/S0014-5793(97)01257-X

学科分类：生物化学/生物物理

来源: John Wiley & Sons Ltd.

【 摘 要 】

Using two synthetic oligonucleotides, we have constructed a new gene containing three zinc finger motifs of the Cys₂-His₂ 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 <sub>d</sub>) comparable to the K <sub>d</sub> 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.

【 授权许可】

Unknown