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Related Experiment Video

Updated: May 13, 2026

Sequence-specific Labeling of Nucleic Acids and Proteins with Methyltransferases and Cofactor Analogues

Sequence-specific Labeling of Nucleic Acids and Proteins with Methyltransferases and Cofactor Analogues

Published on: November 22, 2014

Reversible protein affinity-labelling using bromomaleimide-based reagents.

Ramiz I Nathani¹, Vijay Chudasama, Chris P Ryan

¹Department of Chemistry, University College London, 20 Gordon Street, London, WC1H OAJ, UK.

Organic & Biomolecular Chemistry

|March 7, 2013

Summary

This summary is machine-generated.

This study introduces a novel method for reversible protein biotinylation using bromomaleimide reagents and reductive cleavage. This technique allows for stable biotinylated protein intermediates and quantitative removal under mild conditions, benefiting chemical biology and proteomics research.

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Synthesis and Bioconjugation of Thiol-Reactive Reagents for the Creation of Site-Selectively Modified Immunoconjugates

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Published on: March 6, 2019

Area of Science:

Chemical Biology
Proteomics
Bioconjugation Chemistry

Background:

Protein labeling is crucial for studying protein function.
Existing biotinylation methods may lack reversibility or require harsh conditions.
Developing reversible protein modification strategies is essential for advanced biological research.

Purpose of the Study:

To develop a novel, mild, and efficient method for reversible protein biotinylation.
To demonstrate the stability of intermediate biotinylated proteins.
To enable quantitative removal of biotin tags under physiological conditions.

Main Methods:

Conjugation of proteins with a bromomaleimide-based reagent.
Reductive cleavage of the biotinylation linkage.
Characterization of biotinylated protein stability at physiological temperature and pH 8.0.
Quantitative assessment of biotin removal using a stoichiometric amount of a bis-thiol.

Main Results:

Successfully achieved reversible protein biotinylation.
Biotinylated protein intermediates demonstrated stability under physiological conditions (temperature and pH 8.0).
Quantitative and mild cleavage of biotin was achieved using a bis-thiol reagent.

Conclusions:

The developed method offers a robust approach for reversible protein biotinylation.
This technique is suitable for applications in chemical biology and proteomics.
The mild and quantitative nature of the method makes it broadly applicable.