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Updated: Jun 27, 2026
09:57
A Facile and Efficient Approach for the Production of Reversible Disulfide Cross-linked Micelles
Published on: December 23, 2016
Reprogramming the lipid peroxidation product 4-ONE as a chemoselective cleavable crosslinker.
Zachary E Paikin1, John M Talbott1, Anthony M Ciancone2
1Department of Chemistry, Emory University, Atlanta, GA, USA.
Nature Communications
|June 25, 2026
Summary
This study repurposes 4-oxo-2-nonenal (4-ONE) from a toxic byproduct into a selective crosslinker for cysteine and lysine residues. This novel chemistry enables precise protein mapping and structural analysis, advancing proteomics research.
Area of Science:
- Biochemistry
- Chemical Biology
- Proteomics
Background:
- Oxidative stress generates lipid peroxidation products that modify proteins.
- Defining residue-pair connectivity and structural changes from these modifications is challenging.
Purpose of the Study:
- To repurpose the lipid peroxidation product 4-oxo-2-nonenal (4-ONE) as a chemoselective crosslinking reagent.
- To develop a method for mapping Cys-Lys crosslinks and determining protein structures.
Main Methods:
- A two-step chemoselective pathway involving Michael addition to cysteine followed by cyclization with lysine.
- Development of a workflow named COSMIc (Crosslink Oxidation to Sulfoxide for Mass-Cleavable Interactomics) for MS-cleavable crosslink identification.
- Application of COSMIc to map crosslinks in the human 26S proteasome.
Main Results:
- 4-ONE acts as a stable Cys-Lys covalent crosslinker under physiological conditions.
- The chemistry supports peptide functionalization, macrocyclization, and selective protein modification.
- COSMIc enables site-resolved identification of linked residues and improves confidence in residue-pair assignment.
- Structurally informative crosslinks were detected in the human 26S proteasome.
Conclusions:
- 4-ONE is a versatile metabolite-derived crosslinker for covalent mapping and structural proteomics.
- COSMIc provides a powerful tool for identifying and characterizing protein crosslinks.
- This approach enhances our understanding of protein structure and function in biological systems.
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