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.
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October 2015
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Review Article|
October 09 2015
A designed repeat protein as an affinity capture reagent
Elizabeth B. Speltz;
Elizabeth B. Speltz
*Department of Molecular Biophysics and Biochemistry, Yale University, 266 Whitney Avenue, New Haven, CT 06511, U.S.A.
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Rebecca S.H. Brown;
Rebecca S.H. Brown
*Department of Molecular Biophysics and Biochemistry, Yale University, 266 Whitney Avenue, New Haven, CT 06511, U.S.A.
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Holly S. Hajare;
Holly S. Hajare
*Department of Molecular Biophysics and Biochemistry, Yale University, 266 Whitney Avenue, New Haven, CT 06511, U.S.A.
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Christian Schlieker;
Christian Schlieker
*Department of Molecular Biophysics and Biochemistry, Yale University, 266 Whitney Avenue, New Haven, CT 06511, U.S.A.
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Lynne Regan
Lynne Regan
1
*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.
1To whom correspondence should be addressed (emaillynne.regan@yale.edu).
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Publisher: Portland Press Ltd
Received:
April 30 2015
Online ISSN: 1470-8752
Print ISSN: 0300-5127
© 2015 Authors; published by Portland Press Limited
2015
Biochem Soc Trans (2015) 43 (5): 874–880.
Article history
Received:
April 30 2015
Citation
Elizabeth B. Speltz, Rebecca S.H. Brown, Holly S. Hajare, Christian Schlieker, Lynne Regan; A designed repeat protein as an affinity capture reagent. Biochem Soc Trans 1 October 2015; 43 (5): 874–880. doi: https://doi.org/10.1042/BST20150091
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