A highly controllable, fast, and specific peptide stapling method for installation of both inter- and intra-peptide thioether crosslinks

Existing solutions for installing peptide crosslinks currently require multiple steps and can typically only include a limited range of side chains. These crosslinks are commonly disulfide bonds that are redox sensitive and short lived in biological environments.

University of Utah researchers have developed an enzyme-based system to create more robust thioether crosslinks with a range of amino acids. This method allows for fast one-step peptide stapling and has been used to generate analogs of FDA-approved therapeutics.

This technology is related to U-8158 and U-8159.

Proof of Concept

• Highly controllable, fast, and specific one-step peptide stapling that occurs between native sidechains.
• Thioether crosslink is much more robust than the disulfide crosslink that is naturally found in many peptide compounds.
• Enzymatic crosslinks are fast and can be carried out at room temperature at approximately a neutral pH.

• Eastman K, Mifflin M, Oblad P, et al. (2023). A Promiscuous rSAM Enzyme Enables Diverse Peptide Cross-linking. ACS Bio Med Chem Au. 3(6): 480-493.
• Rush K, Eastman K, Kincannon W, et al. (2023). Peptide Selenocysteine Substitutions Reveal Direct Substrate-Enzyme Interactions at Auxiliary Clusters in Radical S-Adenosyl-l-methionine Maturases. J Am Chem Soc. 145(18): 10167-10177.
• Eastman K, Kincannon W, Bandarian V. (2022) Leveraging Substrate Promiscuity of a Radical S-Adenosyl-L-methionine RiPP Maturase toward Intramolecular Peptide Cross-Linking Applications. ACS Cent Sci. 8 (8): 1209-1217. 

Jason Young
(801) 587-0519
jason.r.young@utah.edu