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OaAEP1-Mediated Enzymatic Synthesis and Immobilization of Polymerized Protein for Single-Molecule Force Spectroscopy
Published on: February 5, 2020
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Modular Diazo Compound for the Bioreversible Late-Stage Modification of Proteins
Journal of the American Chemical Society
|March 15, 2023
Summary
Researchers developed a novel bioreversible protein modification strategy. This method enables efficient, traceless protein delivery into mammalian cells, offering new tools for chemical biology applications.
Area of Science:
- Bioconjugation Chemistry
- Chemical Biology
- Protein Engineering
Background:
- Protein modification is crucial for drug delivery and diagnostics.
- Existing methods often result in irreversible changes or lack efficiency.
- A need exists for versatile, bioreversible protein conjugation strategies.
Purpose of the Study:
- To develop a versatile strategy for the bioreversible modification of proteins.
- To enable chemoselective conjugation and esterase-mediated cleavage for traceless delivery.
- To demonstrate the utility of this strategy for cellular delivery of functional proteins.
Main Methods:
- Synthesis of a tricomponent molecule with diazo, pyridyl disulfide, and self-immolative carbonate groups.
- Chemoselective esterification of protein carboxyl groups.
- Late-stage functionalization with thiolated ligands.
- Esterase-mediated cleavage for bioreversible conjugation.
Main Results:
- Successful generation of protein conjugates (cytochrome c and GFP) with diverse domains.
- Demonstrated cellular delivery of proteins into the cytosol of live mammalian cells, even in serum.
- Showcased functional delivery of cytochrome c, inducing apoptosis, highlighting advantages over irreversible conjugation.
Conclusions:
- The developed strategy offers a versatile and traceless method for bioreversible protein modification.
- This approach facilitates efficient protein delivery into live cells.
- The strategy opens new avenues for chemical biologists in protein engineering and therapeutics.
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