title: A two-component 'double-click' approach to peptide stapling.
abstract: Peptide cyclization is a useful strategy for the stabilization of short flexible peptides into well-defined bioactive conformations, thereby enhancing their ability to interact with proteins and other important biomolecules. We present an optimized procedure for the stabilization of linear diazido peptides in an α-helical conformation upon reaction with dialkynyl linkers under Cu(I) catalysis. As this procedure generates side chain-cyclized peptides bearing a bis-triazole linkage, it is referred to as 'double-click' stapling. Double-click stapling can enhance the binding affinity, proteolytic stability and cellular activity of a peptide inhibitor. A distinguishing feature of double-click stapling is the efficiency with which peptides bearing different staple linkages can be synthesized, thus allowing for modular control over peptide bioactivity. This protocol describes the double-click reaction between a 1,3-dialkynylbenzene linker and peptides that contain azidoornithine. Subsequent peptide purification and confirmation steps are also described. The entire double-click stapling protocol can be completed in ∼48 h, including two overnight lyophilization steps.
authors.affiliation: University Chemical Laboratory, University of Cambridge, Cambridge, UK.
authors.forename: Yu Heng
authors.initials: YH
authors.lastname: Lau
authors.affiliation: University Chemical Laboratory, University of Cambridge, Cambridge, UK.
authors.forename: Yuteng
authors.initials: Y
authors.lastname: Wu
authors.affiliation: University Chemical Laboratory, University of Cambridge, Cambridge, UK.
authors.forename: Peterson
authors.initials: P
authors.lastname: de Andrade
authors.affiliation: University Chemical Laboratory, University of Cambridge, Cambridge, UK.
authors.forename: Warren R J D
authors.initials: WR
authors.lastname: Galloway
authors.affiliation: University Chemical Laboratory, University of Cambridge, Cambridge, UK.
authors.forename: David R
authors.initials: DR
authors.lastname: Spring
mesh_terms: D060326:Chemistry Techniques, Synthetic / Q000295:instrumentation / Q000379:methods*; D002851:Chromatography, High Pressure Liquid / Q000379:methods; D003500:Cyclization; D010455:Peptides / Q000138:chemical synthesis / Q000737:chemistry*; D011487:Protein Conformation; D055550:Protein Stability; D012997:Solvents / Q000737:chemistry
chemical_list: D010455:Peptides; D012997:Solvents