A designed repeat protein as an affinity capture reagent.

Elizabeth B Speltz Rebecca S H Brown Holly S Hajare Christian Schlieker Lynne Regan

Biochem Soc Trans

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.

Published: October 2015

Repeat proteins, especially tetratricopeptide repeat (TPR) proteins, are prime candidates for protein engineering and design due to their unique structures and stability.* -
Previous studies have successfully manipulated TPR protein structures and created various peptide-binding TPR modules, which can be combined into larger assemblies.* -
The current focus demonstrates the practical application of a specific TPR-peptide interaction in affinity purification, showing how fundamental research can result in valuable tools.*

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.

Download full-text PDF	Source
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC5683849	PMC
http://dx.doi.org/10.1042/BST20150091	DOI Listing

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