期刊
ACS PHARMACOLOGY & TRANSLATIONAL SCIENCE
卷 5, 期 10, 页码 859-871出版社
AMER CHEMICAL SOC
DOI: 10.1021/acsptsci.2c00155
关键词
selectivity; chemical probes; chemical proteomics; triple negative breast cancer; USP34; PKMYT1
资金
- Macquarie University [MQRDG9201601650]
- iMQRES Scholarship
The SADL approach utilizes two covalent modifiers under a common ligand directive for precise protein labeling, improving target selectivity and labeling precision, broadening protein identification opportunities, aiding in understanding biological or disease mechanisms, and accelerating drug target discovery.
Covalent modification of endogenous proteins by chemical probes is used for proteome-wide profiling of cellular protein function and drug discovery. However, probe selectivity in the complex cellular environment is a challenge, and new probes with better target selectivity are continuously needed. On the basis of the success of monocovalent activity-based and reactivity based probes, an approach of structurally aligned dual-modifier labeling (SADL) was investigated here on its potential in improving target precision. Two reactive groups, based on the acrylamide and NHS ester chemistry, were linked with structural alignment to be under the same anilinoquinazoline ligand-directive for targeting the epidermal growth factor receptor (EGFR) protein kinase as the model system for proteome-wide profiling. The SADL approach was compared with its monocovalent precursors in a label-free MaxLFQ workflow using MDA-MB-468 triple negative breast cancer cells. The dual-modifier probe consistently showed labeling of EGFR with improved precision over both monocovalent precursors under various controls. The workflow also labeled endogenous USP34 and PKMYT1 with high selectivity. Precision labeling with two covalent modifiers under a common ligand directive may broaden protein identification opportunities in the native environment to complement genetic and antibody-based approaches for elucidating biological or disease mechanisms, as well as accelerating drug target discovery.
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