Cargando…

Exploring the binding of rationally engineered tandem-repeat proteins to E3 ubiquitin ligase Keap1

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 sho...

Descripción completa

Detalles Bibliográficos
Autores principales: Madden, Sarah K, Itzhaki, Laura S
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Oxford University Press 2021
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC8660007/
https://www.ncbi.nlm.nih.gov/pubmed/34882773
http://dx.doi.org/10.1093/protein/gzab027
Descripción
Sumario: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-NH(2,) with low nanomolar affinity for Keap1, highlighting the potential of the repeat-protein class for application in peptide display.