Kinetic Assay of the Michael Addition‐Like Thiol–Ene Reaction and Insight into Protein Bioconjugation |
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| Authors: | Fei‐He Ma Jia‐Liang Chen Qing‐Feng Li Hui‐Hui Zuo Feng Huang Prof Xun‐Cheng Su |
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| Affiliation: | State Key Laboratory of Elemento‐organic Chemistry, Collaborative Innovation Center of Chemical Science and Engineering (Tianjin), Nankai University, Weijin Road 94, Tianjin 300071 (China), Fax: (+86)?22‐23500623 |
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| Abstract: | The chemical modification of proteins is a valuable technique in understanding the functions, interactions, and dynamics of proteins. Reactivity and selectivity are key issues in current chemical modification of proteins. The Michael addition‐like thiol–ene reaction is a useful tool that can be used to tag proteins with high selectivity for the solvent‐exposed thiol groups of proteins. To obtain insight into the bioconjugation of proteins with this method, a kinetic analysis was performed. New vinyl‐substituted pyridine derivatives were designed and synthesized. The reactivity of these vinyl tags with L ‐cysteine was evaluated by UV absorption and high‐resolution NMR spectroscopy. The results show that protonation of pyridine plays a key role in the overall reaction rates. The kinetic parameters were assessed in protein modification. The different reactivities of these vinyl tags with solvent‐exposed cysteine is valuable information in the selective labeling of proteins with multiple functional groups. |
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| Keywords: | conjugation NMR spectroscopy proteins thiol chemistry thiol– ene reaction |
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