Conversion of antennapedia homeodomain to zinc finger-like domain: Zn(II)-induced change in protein conformation and DNA binding.
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.
Hori
Yuichiro
Y
Institute for Chemical Research, Kyoto University, Uji, Japan.
Sugiura
Yukio
Y
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
D051943:Antennapedia Homeodomain Protein; D018398:Homeodomain Proteins; D009687:Nuclear Proteins; D014157:Transcription Factors; D004247:DNA; D015032:Zinc