<i>O</i>-Cyanobenzaldehydes Irreversibly Modify Both Buried and Exposed Lysine Residues in Live Cells

Lysine residue represents an attractive site for covalent drug development due to its high abundance (5.6%) and critical functions. However, very few lysines have been characterized to be accessible to covalent ligands and perturb the protein functions, owing to their protonation state and adjacent steric hindrance. Herein, we report a new lysine bioconjugation chemistry, <i>O</i>-cyanobenzaldehyde (CNBA), that enables selective modification of the lysine ε-amine to form iso-indolinones under physiological conditions. Activity-based proteome profiling enabled the mapping of 3451 lysine residues and 85 endogenous kinases in live cells, highlighting its potential for modifying hyper-reactive lysines within the proteome or buried catalytic lysines within the kinome. Further protein crystallography and mass spectrometry confirmed that K271_ABL1 and K162_AURKA are covalently targetable sites in kinases. Leveraging a structure-based drug design, we incorporated CNBA into the core structure of Nutlin-3 to irreversibly inhibit the MDM2-p53 interaction by targeting an exposed lysine K94 on the surface of murine double minute 2. Importantly, we have demonstrated the potential application of CNBA as a lysine-recognized bioconjugation agent for developing new antibody-drug conjugates. The results collectively validate CNBA as a new selective and efficient modifying agent with broad applications for both buried and exposed lysine residues in live cells.

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Publication Details

Journal
Journal of the American Chemical Society
Published
2025-03-28
DOI
https://doi.org/10.1021/jacs.4c18006
Citations
11
Primary Topic
Click Chemistry and Applications
Type
article
Field-Weighted Citation Impact
10.30

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article

O-Cyanobenzaldehydes Irreversibly Modify Both Buried and Exposed Lysine Residues in Live Cells

Ke Ding, Liping Duan, Yang Zhou, Nan Ma et al.
11 citations
Journal of the American Chemical Society
Click Chemistry and Applications
10.30
article

O-Cyanobenzaldehydes Irreversibly Modify Both Buried and Exposed Lysine Residues in Live Cells

Ke Ding, Liping Duan, Yang Zhou, Nan Ma, Xiaorong Qiu, Xiaomei Ren, Ruijun Tian, Jiangnan Zheng, Yujun Zhao, Jinwei Zhang, Zhimin Zhang, Lin Li, Zhen Wang, Shijie Zhang, Xinling Li, Weixue Huang, Huan Ling
article en
11 citations

Abstract

Lysine residue represents an attractive site for covalent drug development due to its high abundance (5.6%) and critical functions. However, very few lysines have been characterized to be accessible to covalent ligands and perturb the protein functions, owing to their protonation state and adjacent steric hindrance. Herein, we report a new lysine bioconjugation chemistry, <i>O</i>-cyanobenzaldehyde (CNBA), that enables selective modification of the lysine ε-amine to form iso-indolinones under physiological conditions. Activity-based proteome profiling enabled the mapping of 3451 lysine residues and 85 endogenous kinases in live cells, highlighting its potential for modifying hyper-reactive lysines within the proteome or buried catalytic lysines within the kinome. Further protein crystallography and mass spectrometry confirmed that K271_ABL1 and K162_AURKA are covalently targetable sites in kinases. Leveraging a structure-based drug design, we incorporated CNBA into the core structure of Nutlin-3 to irreversibly inhibit the MDM2-p53 interaction by targeting an exposed lysine K94 on the surface of murine double minute 2. Importantly, we have demonstrated the potential application of CNBA as a lysine-recognized bioconjugation agent for developing new antibody-drug conjugates. The results collectively validate CNBA as a new selective and efficient modifying agent with broad applications for both buried and exposed lysine residues in live cells.

Journal of the American Chemical SocietyVol. 147(14)
Jinan University (CN), Chinese Academy of Sciences (CN), Southern University of Science and Technology (CN), Laboratoire de Chémo-biologie synthétique et thérapeutique (FR), Shanghai Institute of Materia Medica (CN), Shanghai Institute of Organic Chemistry (CN), University of Chinese Academy of Sciences (CN)
National Natural Science Foundation of China, Chinese Academy of Sciences, National Key Research and Development Program of China, Basic and Applied Basic Research Foundation of Guangdong Province
Openalex Percentile: Top 2%
Click Chemistry and Applications
10.30
DOISourceOpenAlex
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