Design of Functional Globular β‐Sheet Miniproteins

• Published in: Chembiochem : a European journal of chemical biology 2024-04, Vol.25 (7), p.e202300745-n/a
• Main Authors: Pham, Truc Lam, Thomas, Franziska
• Format: Article
• Language: English
• Subjects: Design; Folding; Metal ions; Nucleic acids; Peptides; Peptides - chemistry; Protein Conformation, beta-Strand; Protein design; Protein Folding; Proteins; Proteins - chemistry; Redox potential; Scaffolds; Temperature; Tryptophan zipper; WW domain; β-hairpin
• ISSN: 1439-4227; 1439-7633
• DOI: 10.1002/cbic.202300745

The design of discrete β‐sheet peptides is far less advanced than e. g. the design of α‐helical peptides. The reputation of β‐sheet peptides as being poorly soluble and aggregation‐prone often hinders active design efforts. Here, we show that this reputation is unfounded. We demonstrate this by looking at the β‐hairpin and WW domain. Their structure and folding have been extensively studied and they have long served as model systems to investigate protein folding and folding kinetics. The resulting fundamental understanding has led to the development of hyperstable β‐sheet scaffolds that fold at temperatures of 100 °C or high concentrations of denaturants. These have been used to design functional miniproteins with protein or nucleic acid binding properties, in some cases with such success that medical applications are conceivable. The β‐sheet scaffolds are not always completely rigid, but can be specifically designed to respond to changes in pH, redox potential or presence of metal ions. Some engineered β‐sheet peptides also exhibit catalytic properties, although not comparable to those of natural proteins. Previous reviews have focused on the design of stably folded and non‐aggregating β‐sheet sequences. In our review, we now also address design strategies to obtain functional miniproteins from β‐sheet folding motifs. Not just the beta versions. Miniprotein design has long been dominated by the well‐understood α‐helical coiled coil scaffolds. With increasing understanding, globular β‐sheet scaffolds, which are generally considered to be prone to aggregation, are also becoming accessible. We review the design of β‐hairpins and WW domains and discuss strategies for engineering these scaffolds into receptors, molecular switches and catalysts.