Building and Breaking Bonds via a Compact S‐Propargyl‐Cysteine to Chemically Control Enzymes and Modify Proteins. Issue 39 (5th September 2018)

• Record Type: Journal Article
• Authors: Liu, Jun; Cheng, Rujin; Wu, Haifan; Li, Shanshan; Wang, Peng G.; DeGrado, William F.; Rozovsky, Sharon; Wang, Lei
• Is Part Of: Angewandte Chemie international edition. Volume 57:Issue 39(2018)
• Page Start: 12702
• Page End: 12706
• Publication Date: 2018-09-05
• Subjects: palladium-mediated cleavage -- propargyl cysteine -- reversible protein modification -- Sonogashira coupling -- thiol–yne; Chemistry -- Periodicals; 540
• DOI: 10.1002/anie.201806197
• Languages: English
• ISSNs: 1433-7851

Abstract

Abstract: Analogous to reversible post‐translational protein modifications, the ability to attach and subsequently remove modifications on proteins would be valuable for protein and biological research. Although bioorthogonal functionalities have been developed to conjugate or cleave protein modifications, they are introduced into proteins on separate residues and often with bulky side chains, limiting their use to one type of control and primarily protein surface. Here we achieved dual control on one residue by genetically encoding S‐propargyl‐cysteine (SprC), which has bioorthogonal alkyne and propargyl groups in a compact structure, permitting usage in protein interior in addition to surface. We demonstrated its incorporation at the dimer interface of glutathione transferase for in vivo crosslinking via thiol–yne click chemistry, and at the active site of human rhinovirus 3C protease for masking and then turning on enzyme activity via Pd‐cleavage of SprC into Cys. In addition, we installed biotin onto EGFP via Sonogashira coupling of SprC and then tracelessly removed it via Pd cleavage. SprC is small in size, commercially available, nontoxic, and allows for bond building and breaking on a single residue. Genetically encoded SprC will be valuable for chemically controlling proteins with an essential Cys and for reversible protein modifications. Abstract : S‐propargyl‐cysteine (SprC) was genetically encoded into proteins at the interface to cross‐link proteins and at the active site to mask an enzyme, which was chemically activated by Pd(TPPTS)4 . In addition, molecular modifications were installed onto proteins via Sonogashira cross‐coupling on SprC by Pd(NO3 )2, and then tracelessly removed with Pd(TPPTS)4 .