title: Redesign of a WW domain peptide for selective recognition of single-stranded DNA.
abstract: A β-sheet miniprotein based on the FBP11 WW1 domain sequence has been redesigned for the molecular recognition of ssDNA. A previous report showed that a β-hairpin peptide dimer, (WKWK)(2), binds ssDNA with low micromolar affinity but with little selectivity over duplex DNA. This report extends those studies to a three-stranded β-sheet miniprotein designed to mimic the OB-fold. The new peptide binds ssDNA with low micromolar affinity and shows about 10-fold selectivity for ssDNA over duplex DNA. The redesigned peptide no longer binds its native ligand, the polyproline helix, confirming that the peptide has been redesigned for the function of binding ssDNA. Structural studies provide evidence that this peptide consists of a well-structured β-hairpin made of strands 2 and 3 with a less structured first strand that provides affinity for ssDNA but does not improve the stability of the full peptide. These studies provide insight into protein-DNA interactions as well as a novel example of protein redesign.
authors.affiliation: Department of Chemistry, CB 3290, University of North Carolina, Chapel Hill, North Carolina 27599, United States.
authors.forename: Amanda L
authors.initials: AL
authors.lastname: Stewart
authors.forename: Jessica H
authors.initials: JH
authors.lastname: Park
authors.forename: Marcey L
authors.initials: ML
authors.lastname: Waters
mesh_terms: D002352:Carrier Proteins / Q000737:chemistry* / Q000235:genetics; D002942:Circular Dichroism; D004277:DNA, Single-Stranded / Q000378:metabolism*; D004268:DNA-Binding Proteins / Q000737:chemistry / Q000378:metabolism*; D019281:Dimerization; D005454:Fluorescence Polarization; D006801:Humans; D007700:Kinetics; D008024:Ligands; D050505:Mutant Proteins / Q000737:chemistry / Q000378:metabolism; D019906:Nuclear Magnetic Resonance, Biomolecular; D010446:Peptide Fragments / Q000737:chemistry / Q000378:metabolism*; D010455:Peptides / Q000378:metabolism; D011489:Protein Denaturation; D015202:Protein Engineering; D054730:Protein Interaction Domains and Motifs; D058849:Protein Refolding; D055550:Protein Stability; D013696:Temperature
chemical_list: D002352:Carrier Proteins; D004277:DNA, Single-Stranded; D004268:DNA-Binding Proteins; D008024:Ligands; D050505:Mutant Proteins; C491404:PRPF40A protein, human; D010446:Peptide Fragments; D010455:Peptides; C011083:polyproline