Fast and Cysteine-Specific Modification of Peptides, Proteins and Bacteriophage Using Chlorooximes
Fa-Jie Chen1, Mengmeng Zheng1, Vincent Nobile1
1Department of Chemistry Boston College, Merkert Chemistry Center, 2609 Beacon Street, Chestnut Hill, MA 02467, USA.
Chemistry (Weinheim an Der Bergstrasse, Germany)
|February 15, 2022
Summary
This study introduces a new chlorooxime method for modifying peptides and proteins, specifically targeting cysteine residues. This efficient and stable conjugation technique was applied to create modified phage libraries.
Area of Science:
- Bioconjugation Chemistry
- Protein Engineering
- Chemical Biology
Background:
- Native peptide and protein modification is crucial for various applications.
- Existing methods often require harsh conditions or lack specificity.
- Cysteine residues offer unique opportunities for targeted chemical modification.
Purpose of the Study:
- To develop a novel, mild, and selective method for cysteine conjugation.
- To enable efficient Cys-Cys stapling in peptides and proteins.
- To demonstrate the utility of this method in constructing modified phage libraries.
Main Methods:
- Utilizing a novel chlorooxime reagent for peptide and protein modification.
- Investigating reaction kinetics and selectivity for cysteine residues.
- Synthesizing a bis-chlorooxime reagent for Cys-Cys stapling.
- Applying the method to bacteriophage for library construction.
Main Results:
- Achieved fast reaction kinetics (k2 = 306±4 M−1 s−1 for GSH) and high selectivity for cysteine.
- Demonstrated high stability of conjugates against acid, base, and thiol nucleophiles.
- Successfully performed Cys-Cys stapling using a bis-chlorooxime reagent.
- Constructed chemically modified phage libraries using the developed method.
Conclusions:
- The chlorooxime-mediated modification is a robust and efficient technique for cysteine conjugation and stapling.
- This method offers a valuable tool for creating modified peptides, proteins, and phage libraries under physiological conditions.
- The developed chemistry provides enhanced stability and selectivity, broadening its applicability in chemical biology and protein engineering.
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