title: A designed repeat protein as an affinity capture reagent.
abstract: Repeat proteins are an attractive target for protein engineering and design. We have focused our attention on the design and engineering of one particular class: tetratricopeptide repeat (TPR) proteins. In previous work, we have shown that the structure and stability of TPR proteins can be manipulated in a rational fashion [Cortajarena (2011) Prot. Sci. 20: , 1042-1047; Main (2003) Structure 11: , 497-508]. Building on those studies, we have designed and characterized a number of different peptide-binding TPR modules and we have also assembled these modules into supramolecular arrays [Cortajarena (2009) ACS Chem. Biol. 5: , 545-552; Cortajarena (2008) ACS Chem. Biol. 3: , 161-166; Jackrel (2009) Prot. Sci. 18: , 762-774; Kajander (2007) Acta Crystallogr. D Biol. Crystallogr. 63: , 800-811]. Here we focus on the development of one such TPR-peptide interaction for a practical application, affinity purification. We illustrate the general utility of our designed protein interaction. Furthermore, this example highlights how basic research on protein-peptide interactions can lead to the development of novel reagents with important practical applications.
authors.affiliation: Department of Molecular Biophysics and Biochemistry, Yale University, 266 Whitney Avenue, New Haven, CT 06511, U.S.A.
authors.forename: Elizabeth B
authors.initials: EB
authors.lastname: Speltz
authors.affiliation: Department of Molecular Biophysics and Biochemistry, Yale University, 266 Whitney Avenue, New Haven, CT 06511, U.S.A.
authors.forename: Rebecca S H
authors.initials: RS
authors.lastname: Brown
authors.affiliation: Department of Molecular Biophysics and Biochemistry, Yale University, 266 Whitney Avenue, New Haven, CT 06511, U.S.A.
authors.forename: Holly S
authors.initials: HS
authors.lastname: Hajare
authors.affiliation: Department of Molecular Biophysics and Biochemistry, Yale University, 266 Whitney Avenue, New Haven, CT 06511, U.S.A.
authors.forename: Christian
authors.initials: C
authors.lastname: Schlieker
authors.affiliation: Department of Molecular Biophysics and Biochemistry, Yale University, 266 Whitney Avenue, New Haven, CT 06511, U.S.A. Integrated Graduate Program in Physical and Engineering Biology, Yale University, 266 Whitney Avenue, New Haven, CT 06511, U.S.A. Department of Chemistry, Yale University, 266 Whitney Avenue, New Haven, CT 06511, U.S.A. lynne.regan@yale.edu.
authors.forename: Lynne
authors.initials: L
authors.lastname: Regan
mesh_terms: D000818:Animals; D002460:Cell Line; D002846:Chromatography, Affinity; D006801:Humans; D055767:Immobilized Proteins / Q000737:chemistry / Q000235:genetics / Q000378:metabolism; D007202:Indicators and Reagents / Q000737:chemistry* / Q000378:metabolism; D008024:Ligands; D010446:Peptide Fragments / Q000737:chemistry / Q000235:genetics / Q000378:metabolism; D015202:Protein Engineering; D054730:Protein Interaction Domains and Motifs; D011993:Recombinant Fusion Proteins / Q000737:chemistry* / Q000235:genetics / Q000378:metabolism; D020449:Repetitive Sequences, Amino Acid
chemical_list: D055767:Immobilized Proteins; D007202:Indicators and Reagents; D008024:Ligands; D010446:Peptide Fragments; D011993:Recombinant Fusion Proteins
keywords: affinity reagent; antibody; protein design; protein purification; protein–protein interactions; repeat protein