Fast and Cysteine-Specific Modification of Peptides,Proteins and Bacteriophage Using Chlorooximes |
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Authors: | Dr Fa-Jie Chen Dr Mengmeng Zheng Vincent Nobile Dr Jianmin Gao |
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Institution: | 1. Department of Chemistry Boston College, Merkert Chemistry Center, 2609 Beacon Street, Chestnut Hill, MA 02467 USA;2. Department of Chemistry Boston College, Merkert Chemistry Center, 2609 Beacon Street, Chestnut Hill, MA 02467 USA
These authors contributed equally to this work. |
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Abstract: | This work reports a novel chlorooxime mediated modification of native peptides and proteins under physiologic conditions. This method features fast reaction kinetics (apparent k2=306±4 M?1s?1 for GSH) and exquisite selectivity for cysteine residues. This cysteine conjugation reaction can be carried out with just single-digit micromolar concentrations of the labeling reagent. The conjugates show high stability towards acid, base, and external thiol nucleophiles. A nitrile oxide species generated in situ is likely involved as the key intermediate. Furthermore, a bis-chlorooxime reagent is synthesized to enable facile Cys-Cys stapling in native peptides and proteins. This highly efficient cysteine conjugation and stapling was further implemented on bacteriophage to construct chemically modified phage libraries. |
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Keywords: | chlorooxime cysteine modification peptide stapling phage modification protein modification |
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