Exploring the binding of rationally engineered tandem-repeat proteins to E3 ubiquitin ligase Keap1.
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Publication Date
2021-02-15Journal Title
Protein Eng Des Sel
ISSN
1741-0126
Publisher
Oxford University Press (OUP)
Language
eng
Type
Article
This Version
VoR
Metadata
Show full item recordCitation
Madden, S. K., & Itzhaki, L. S. (2021). Exploring the binding of rationally engineered tandem-repeat proteins to E3 ubiquitin ligase Keap1.. Protein Eng Des Sel https://doi.org/10.1093/protein/gzab027
Abstract
The process of displaying functional peptides by 'grafting' them onto loops of a stable protein scaffold can be used to impart binding affinity for a target, but it can be difficult to predict the affinity of the grafted peptide and the effect of grafting on scaffold stability. In this study, we show that a series of peptides that bind to the E3 ubiquitin ligase Keap1 can be grafted into the inter-repeat loop of a consensus-designed tetratricopeptide repeat (CTPR) protein resulting in proteins with high stability. We found that these CTPR-grafted peptides had similar affinities to their free peptide counterparts and achieved a low nanomolar range. This result is likely due to a good structural match between the inter-repeat loop of the CTPR and the Keap1-binding peptide. The grafting process led to the discovery of a new Keap1-binding peptide, Ac-LDPETGELL-NH2, with low nanomolar affinity for Keap1, highlighting the potential of the repeat-protein class for application in peptide display.
Keywords
NRF2, Biologics, Tetratricopeptide Repeat, Peptide Grafting, Tandem-repeat Protein
Sponsorship
CRUK Pioneer Award (C17838/A22676)
CRUK Biotherapeutic Drug Discovery Project Award (C17838/A27225). BBSRC Doctoral Training Scholarship (BB/J014540/1
Funder references
Cancer Research UK (22676)
Cancer Research UK (27225)
Biotechnology and Biological Sciences Research Council (BB/J014540/1)
Identifiers
34882773, PMC8660007
External DOI: https://doi.org/10.1093/protein/gzab027
This record's URL: https://www.repository.cam.ac.uk/handle/1810/333107
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