A Chemoselective Rapid Azo-Coupling Reaction (CRACR) for Unclickable Bioconjugation

Chemoselective modification of complex biomolecules has become a cornerstone of chemical biology. Despite the exciting developments of the past two decades, the demand for new chemoselective reactions with unique abilities, and those compatible with existing chemistries for concurrent multisite-dire...

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Bibliographic Details
Published in:Journal of the American Chemical Society Vol. 139; no. 34; pp. 11670 - 11673
Main Authors: Addy, Partha Sarathi, Erickson, Sarah B., Italia, James S., Chatterjee, Abhishek
Format: Journal Article
Language:English
Published: WASHINGTON Amer Chemical Soc 30.08.2017
Subjects:
5-Hydroxytryptophan - chemistry
Animals
Azo Compounds - chemistry
Bacterial Proteins - chemistry
Cell Line
Chemistry
Chemistry, Multidisciplinary
Click Chemistry - methods
Escherichia coli - chemistry
Fluorescent Dyes - chemistry
Humans
Immunoconjugates - chemistry
Indoles - chemistry
Models, Molecular
Physical Sciences
Proteins - chemistry
Science & Technology
Staining and Labeling - methods
AZIDE-ALKYNE CYCLOADDITION
BIOORTHOGONAL REACTIONS
TOBACCO-MOSAIC-VIRUS
GENETIC-CODE
CLICK CHEMISTRY
SURFACE MODIFICATION
UNNATURAL AMINO-ACIDS
MAMMALIAN-CELLS
SELECTIVE MODIFICATION
TRANS-CYCLOOCTENES
ISSN:0002-7863, 1520-5126, 1520-5126
Online Access:Get more information
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Summary:Chemoselective modification of complex biomolecules has become a cornerstone of chemical biology. Despite the exciting developments of the past two decades, the demand for new chemoselective reactions with unique abilities, and those compatible with existing chemistries for concurrent multisite-directed labeling, remains high. Here we show that 5-hydroxyindoles exhibit remarkably high reactivity toward aromatic diazonium ions and this reaction can be used to chemoselectively label proteins. We have previously genetically encoded the noncanonical amino acid 5-hydroxytryptophan in both E. coli and eitkaryotes, enabling efficient site-specific incorporation of S-hydroxyindole into virtually any protein. The S-hydroxytryptophan residue was shown to allow rapid; chemoselective protein modification using the azocoupling reaction, and the utility of this bioconjugation strategy was further illustrated by generating a functional antibody fluorophore conjugate. Although the resulting azo-linkage is otherwise stable, we show that it can be efficiently cleaved upon treatment with dithionite. Our work establishes a unique chemoselective "unclickable" bioconjugation strategy to site-specifically modify proteins expressed in both bacteria and eukaryotes.
Bibliography:NIH RePORTER
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ISSN:0002-7863
1520-5126
1520-5126
DOI:10.1021/jacs.7b05125