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Updated: Sep 9, 2025
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Chemoselective Modification of Viral Surfaces via Bioorthogonal Click Chemistry
Published on: August 19, 2012
24.8K
Non-Hydrolysable ADP-Ribosylated SUMO2 via Copper(I) Catalyzed Azide Alkyne Cycloaddition
Matthew D Heijne1, Gerbrand J van der Heden van Noort2
1Department of Cell and Chemical Biology, Leiden University Medical Centre, Leiden, The Netherlands.
Methods in Molecular Biology (Clifton, N.J.)
|August 28, 2025
Summary
Researchers synthesized a stable ADP-ribose linkage to study protein interactions. This method stabilizes ADP-ribosylated proteins, preventing degradation and aiding in cellular localization and pulldown-enrichment experiments.
Area of Science:
- Biochemistry
- Chemical Biology
- Molecular Biology
Background:
- ADP-ribosylation is a post-translational modification catalyzed by ADPr-transferases using β-NAD+.
- The α-glycosidic bond formed is labile and susceptible to cellular degradation, hindering study.
- Stabilizing this bond is crucial for investigating ADP-ribosylated protein localization, interactions, and identification.
Purpose of the Study:
- To develop a method for stabilizing the ADP-ribose linkage in proteins.
- To create a non-hydrolyzable analog of ADP-ribosylated proteins for biochemical studies.
- To facilitate the study of ADP-ribosylation dynamics and interactions within cellular systems.
Main Methods:
- Synthesis of a stable triazole linkage mimicking the ADP-ribose bond via copper(I) catalyzed azide-alkyne cycloaddition (CuAAC).
- Utilized 1″-α-azido-ADPribose and N-terminally biotinylated SUMO21-92 with a propargyl glycine substitution at His17.
- Employed click chemistry to covalently link the modified ADP-ribose analog to a protein.
Main Results:
- Successfully synthesized a stable, non-hydrolyzable triazole linkage at the ADP-ribose site.
- Demonstrated the feasibility of using CuAAC for creating stabilized ADP-ribosylated protein analogs.
- Established a method for generating biotinylated, stabilized ADP-ribosylated protein for pulldown assays.
Conclusions:
- The developed method provides a stable tool for studying ADP-ribosylation.
- This approach enables robust investigation of ADP-ribosylated protein localization and interactions.
- The stabilized analog is valuable for identifying interacting partners and understanding the biological roles of ADP-ribosylation.
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