A Comparative Study of Bioorthogonal Reactions with Azides

Detection of metabolites and post-translational modifications can be achieved using the azide as a bioorthogonal chemical reporter. Once introduced into target biomolecules, either metabolically or through chemical modification, the azide can be tagged with probes using one of three highly selective...

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Bibliographic Details
Published in:ACS chemical biology 2006-11, Vol.1 (10), p.644-648
Main Authors: Agard, Nicholas J, Baskin, Jeremy M, Prescher, Jennifer A, Lo, Anderson, Bertozzi, Carolyn R
Format: Article
Language:English
Subjects:
Azides - chemistry
Blotting, Western
Catalysis
Chemistry, Pharmaceutical - methods
Copper - chemistry
Drug Design
Escherichia coli - metabolism
Kinetics
Models, Chemical
Peptides - chemistry
Protein Binding
Proteins - chemistry
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Summary:Detection of metabolites and post-translational modifications can be achieved using the azide as a bioorthogonal chemical reporter. Once introduced into target biomolecules, either metabolically or through chemical modification, the azide can be tagged with probes using one of three highly selective reactions: the Staudinger ligation, the Cu(I)-catalyzed azide-alkyne cycloaddition, or the strain-promoted [3 + 2] cycloaddition. Here, we compared these chemistries in the context of various biological applications, including labeling of biomolecules in complex lysates and on live cell surfaces. The Cu(I)-catalyzed reaction was found to be most efficient for detecting azides in protein samples but was not compatible with live cells due to the toxicity of the reagents. Both the Staudinger ligation and the strain-promoted [3 + 2] cycloaddition using optimized cyclooctynes were effective for tagging azides on live cells. The best reagent for this application was dependent upon the specific structure of the azide. These results provide a guide for biologists in choosing a suitable ligation chemistry.
ISSN:1554-8929
1554-8937
DOI:10.1021/cb6003228