title: Conversion of antennapedia homeodomain to zinc finger-like domain: Zn(II)-induced change in protein conformation and DNA binding.
abstract: From the standpoint of protein dynamics and metalloprotein design, it is interesting to create an artificial protein which induces structural change and regulates its function by metal-ion binding. We engineered a novel protein, "Antennafinger (Ant-F)", whose structure and function can be controlled with Zn(II), by introducing the consensus sequence of a Cys(2)His(2)-type zinc finger protein into a non-metalloprotein scaffold, an Antennapedia homeodomain mutant (Ant-wt), selected using a motif-searching system. The circular dichroism studies demonstrate that Ant-F has secondary structures similar to Ant-wt and also changes its conformation due to Zn(II)-binding. The optical absorption spectra of the Co(II) complexes of Ant-F and its derivative proteins suggest that the geometry of the metal center of holo-Ant-F is tetrahedral and that the mutated Cys(2)His(2) residues are involved in the complex formation. In addition, the gel mobility shift assay reveals that the DNA binding activity of Ant-F can be regulated through Zn(II)-induced structural alteration. These results provide valuable information about the dynamic properties of proteins and a novel concept for metalloprotein design.
authors.affiliation: Institute for Chemical Research, Kyoto University, Uji, Japan.
authors.forename: Yuichiro
authors.initials: Y
authors.lastname: Hori
authors.forename: Yukio
authors.initials: Y
authors.lastname: Sugiura
mesh_terms: D000595:Amino Acid Sequence; D051943:Antennapedia Homeodomain Protein; D002942:Circular Dichroism; D004247:DNA / Q000378:metabolism*; D018398:Homeodomain Proteins / Q000737:chemistry* / Q000378:metabolism; D008969:Molecular Sequence Data; D009687:Nuclear Proteins; D011487:Protein Conformation; D017386:Sequence Homology, Amino Acid; D014157:Transcription Factors; D015032:Zinc / Q000378:metabolism*; D016335:Zinc Fingers
chemical_list: D051943:Antennapedia Homeodomain Protein; D018398:Homeodomain Proteins; D009687:Nuclear Proteins; D014157:Transcription Factors; D004247:DNA; D015032:Zinc