Engineering Stable Miniproteins: The Journey from Computational Design to Tertiary Structure Stability

Recent advances in miniproteins containing -amino acid residues have changed the understanding and manipulation of helical structures, facilitated by the use of cycloalkane-based -building blocks.These /-peptides exhibit unique structural features and have demonstrated promising biological activities, such as antimicrobial properties and protein interaction inhibition 1 .Additionally, miniproteins containing -residues, have been studied extensively for their well-defined three-dimensional structures, making them attractive candidates for various applications due to their precise conformation control and resistance to enzymatic proteolysis 2 .While secondary structure studies have been successful, research on more extended systems is limited, especially of peptide foldamers composed of cis-aminocyclopentanecarboxylic acid.Initially, miniproteins were designed by naturally excreting proteins.Since then, however, the field has evolved into a rational design approach that respects not only biochemical and physicochemical principles but also empirical data.Nowadays, the use of computational methods that rely on fragment-based design is crucial in de novo design of most miniproteins 3 .