title: Alpha-helix substitution: novel approach for the design of a new zinc finger peptides for AT-rich sequence.
abstract: A novel strategy for the design of a zinc finger peptide on the basis of alpha-helix substitution has been demonstrated. Sp1HM is a helix-substituted mutant for the wild-type Sp1(zf123) and its alpha-helix of each finger is replaced by that of fingers 4-6 of CF2-II. The circular dichroism spectrum of Sp1HM suggests that Sp1HM has an ordered secondary structure similar to Sp1(zf123). From the analyses of the DNA binding affinity and specificity by gel mobility shift assay, it is clearly indicated that Sp1HM specifically binds to the AT-rich sequence (5'-GTA TAT ATA-3') with 3 nM dissociation constants. Moreover, the zinc finger peptides for the sequence alternating between the AT- and GC-rich subsites can also be created by the alpha-helix substitution. This strategy is evidently effective and is also more convenient than the phage display method. Consequently, our design method is widely applicable to creating zinc finger peptides with novel binding specificities.
authors.affiliation: Institute for Chemical Research, Kyoto University, Gokasho, Uji 611-0011, Japan.
authors.forename: Makoto
authors.initials: M
authors.lastname: Nagaoka
authors.forename: Yoshihide
authors.initials: Y
authors.lastname: Doi
authors.forename: Jun
authors.initials: J
authors.lastname: Kuwahara
authors.forename: Yukio
authors.initials: Y
authors.lastname: Sugiura
mesh_terms: D000225:Adenine / Q000737:chemistry*; D000595:Amino Acid Sequence; D001483:Base Sequence; D017931:DNA Primers; D008969:Molecular Sequence Data; D017433:Protein Structure, Secondary; D017386:Sequence Homology, Amino Acid; D013941:Thymine / Q000737:chemistry*; D016335:Zinc Fingers
chemical_list: D017931:DNA Primers; D000225:Adenine; D013941:Thymine