title: Application of Thiol-yne/Thiol-ene Reactions for Peptide and Protein Macrocyclizations.
abstract: The application of thiol-yne/thiol-ene reactions to synthesize mono- and bicyclic-stapled peptides and proteins is reported. First, a thiol-ene-based peptide-stapling method in aqueous conditions was developed. This method enabled the efficient stapling of recombinantly expressed coil-coiled proteins. The resulting stapled protein demonstrated higher stability in its secondary structure than the unstapled version. Furthermore, a thiol-yne coupling was performed by using an α,ω-diyne to react with two cysteine residues to synthesize a stapled peptide with two vinyl sulfide groups. The stapled peptide could further react with another biscysteine peptide to yield a bicyclic stapled peptide with enhanced properties. For example, the cell permeability of a stapled peptide was further increased by appending an oligoarginine cell-penetrating peptide. The robustness and versatility of thiol-yne/thiol-ene reactions that can be applied to both synthetic and expressed peptides and proteins were demonstrated.
authors.affiliation: Department of Biochemistry, University of Utah, 15 N, Medical Drive East 4100, Salt Lake City, UT, 84112, USA.
authors.forename: Yuanxiang
authors.initials: Y
authors.lastname: Wang
authors.affiliation: Department of Pharmaceutics and Pharmaceutical Chemistry, University of Utah, 30 S 2000 E, Rm 2916, Salt Lake City, UT, 84112, USA.
authors.forename: Benjamin J
authors.initials: BJ
authors.lastname: Bruno
authors.affiliation: Department of Medicinal Chemistry, University of Utah, 30 S 2000 E, Rm 4914, Salt Lake City, UT, 84112, USA.
authors.forename: Sean
authors.initials: S
authors.lastname: Cornillie
authors.affiliation: Department of Biochemistry, University of Utah, 15 N, Medical Drive East 4100, Salt Lake City, UT, 84112, USA.
authors.forename: Jason M
authors.initials: JM
authors.lastname: Nogieira
authors.affiliation: Department of Biochemistry, University of Utah, 15 N, Medical Drive East 4100, Salt Lake City, UT, 84112, USA.
authors.forename: Diao
authors.initials: D
authors.lastname: Chen
authors.affiliation: Department of Medicinal Chemistry, University of Utah, 30 S 2000 E, Rm 4914, Salt Lake City, UT, 84112, USA.
authors.forename: Thomas E
authors.initials: TE
authors.lastname: Cheatham
authors.affiliation: Department of Pharmaceutics and Pharmaceutical Chemistry, University of Utah, 30 S 2000 E, Rm 2916, Salt Lake City, UT, 84112, USA.
authors.forename: Carol S
authors.identifier: http://orcid.org/0000-0001-8409-0151
authors.initials: CS
authors.lastname: Lim
authors.affiliation: Department of Biochemistry, University of Utah, 15 N, Medical Drive East 4100, Salt Lake City, UT, 84112, USA.
authors.forename: Danny Hung-Chieh
authors.identifier: http://orcid.org/0000-0001-9110-614X
authors.initials: DH
authors.lastname: Chou
mesh_terms: D000595:Amino Acid Sequence; D057846:Cell-Penetrating Peptides / Q000737:chemistry*; D002850:Chromatography, Gel; D003500:Cyclization; D003545:Cysteine / Q000737:chemistry; D017433:Protein Structure, Secondary; D013438:Sulfhydryl Compounds / Q000737:chemistry*; D013440:Sulfides / Q000737:chemistry*
chemical_list: D057846:Cell-Penetrating Peptides; D013438:Sulfhydryl Compounds; D013440:Sulfides; D003545:Cysteine
keywords: peptide macrocyclization; peptide stapling; thiol-ene coupling; thiol-yne coupling