title: Engineering a protein scaffold from a PHD finger.
abstract: The design of proteins with tailored functions remains a relatively elusive goal. Small size, a well-defined structure, and the ability to maintain structural integrity despite multiple mutations are all desirable properties for such designer proteins. Many zinc binding domains fit this description. We determined the structure of a PHD finger from the transcriptional cofactor Mi2beta and investigated the suitability of this domain as a scaffold for presenting selected binding functions. The two flexible loops in the structure were mutated extensively by either substitution or expansion, without affecting the overall fold of the domain. A binding site for the corepressor CtBP2 was also grafted onto the domain, creating a new PHD domain that can specifically bind CtBP2 both in vitro and in the context of a eukaryotic cell nucleus. These results represent a step toward designing new regulatory proteins for modulating aberrant gene expression in vivo.
authors.affiliation: School of Molecular and Microbial Biosciences, University of Sydney, Sydney, New South Wales 2006 Australia.
authors.forename: Ann H Y
authors.initials: AH
authors.lastname: Kwan
authors.forename: David A
authors.initials: DA
authors.lastname: Gell
authors.forename: Alexis
authors.initials: A
authors.lastname: Verger
authors.forename: Merlin
authors.initials: M
authors.lastname: Crossley
authors.forename: Jacqueline M
authors.initials: JM
authors.lastname: Matthews
authors.forename: Joel P
authors.initials: JP
authors.lastname: Mackay
mesh_terms: D000595:Amino Acid Sequence; D002942:Circular Dichroism; D008969:Molecular Sequence Data; D009154:Mutation; D011487:Protein Conformation; D015202:Protein Engineering; D017386:Sequence Homology, Amino Acid; D013054:Spectrophotometry, Atomic; D014157:Transcription Factors / Q000737:chemistry*
chemical_list: D014157:Transcription Factors