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Traceless Click-Assisted Native Chemical Ligation Enabled by Protecting Dibenzocyclooctyne from Acid-Mediated Rearrangement with Copper(I)
- Source :
- Bioconjug Chem
- Publication Year :
- 2021
-
Abstract
- The scope of proteins accessible to total chemical synthesis via native chemical ligation (NCL) is often limited by slow ligation kinetics. Here we describe Click-Assisted NCL (CAN), in which peptides are incorporated with traceless “helping hand” lysine linkers that enable addition of dibenzocyclooctyne (DBCO) and azide handles. The resulting strain-promoted alkyne-azide cycloaddition (SPAAC) increases their effective concentration to greatly accelerate ligations. We demonstrate that copper(I) protects DBCO from acid-mediated rearrangement during acidic peptide cleavage, enabling direct production of DBCO synthetic peptides. Excitingly, triazole-linked model peptides ligated rapidly and accumulated little side product due to the fast reaction time. Using the E. coli ribosomal subunit L32 as a model protein, we further demonstrate that SPAAC, ligation, desulfurization, and linker cleavage steps can be performed in one pot. CAN is a useful method for overcoming challenging ligations involving sterically hindered junctions. Additionally, CAN is anticipated to be an important stepping stone towards a multi-segment, one-pot, templated ligation system.
- Subjects :
- Pharmacology
Azides
Chemistry
Organic Chemistry
Kinetics
Biomedical Engineering
food and beverages
Pharmaceutical Science
Bioengineering
Cleavage (embryo)
Native chemical ligation
Chemical synthesis
Combinatorial chemistry
Cycloaddition
Article
chemistry.chemical_compound
Alkynes
Escherichia coli
Click Chemistry
Azide
Linker
Copper-free click chemistry
Copper
Biotechnology
Subjects
Details
- ISSN :
- 15204812
- Volume :
- 32
- Issue :
- 10
- Database :
- OpenAIRE
- Journal :
- Bioconjugate chemistry
- Accession number :
- edsair.doi.dedup.....a01809f0223fb33b0a28024e989f96fa