Tunable Heteroaromatic Sulfones Enhance in-Cell Cysteine Profiling
| Autor: | Brittany L Stinger, Bruce A. Palfey, Brent R. Martin, Yu‐hsuan Kuo, Hashim F. Motiwala |
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| Rok vydání: | 2019 |
| Předmět: |
chemistry.chemical_classification
Alkyne General Chemistry Alkylation 010402 general chemistry Mass spectrometry 01 natural sciences Biochemistry Combinatorial chemistry Catalysis 0104 chemical sciences Matrix-assisted laser desorption/ionization Colloid and Surface Chemistry chemistry Nucleophilic aromatic substitution Alkynes Molecular Probes Spectrometry Mass Matrix-Assisted Laser Desorption-Ionization Electrophile Thiol Indicators and Reagents Cysteine Sulfones Oxidation-Reduction |
| Zdroj: | Journal of the American Chemical Society. 142(4) |
| ISSN: | 1520-5126 |
| Popis: | Heteroaromatic sulfones react with cysteine via nucleophilic aromatic substitution, providing a mechanistically selective and irreversible scaffold for cysteine conjugation. Here we evaluate a library of heteroaromatic sulfides with different oxidation states, heteroatom substitutions, and a series of electron-donating and electron-withdrawing substituents. Select substitutions profoundly influence reactivity and stability compared to conventional cysteine conjugation reagents, increasing the reaction rate by >3 orders of magnitude. The findings establish a series of synthetically accessible electrophilic scaffolds tunable across multiple centers. New electrophiles and their corresponding alkyne conjugates were profiled directly in cultured cells, achieving thiol saturation in a few minutes at submillimolar concentrations. Direct addition of desthiobiotin-functionalized probes to cultured cells simplified enrichment and elution to enable the mass spectrometry discovery of >3000 reactive and/or accessible thiols labeled in their native cellular environments in a fraction of the standard analysis time. Surprisingly, only half of the annotated cysteines were identified by both iodoacetamide-desthiobiotin and methylsulfonylbenzothiazole-desthiobiotin in replicate experiments, demonstrating complementary detection by mass spectrometry analysis. These probes offer advantages over existing cysteine alkylation reagents, including accelerated reaction rates, improved stability, and robust ionization for mass spectrometry applications. Overall, heteroaromatic sulfones provide modular tunability, shifted chromatographic elution times, and superior in-cell cysteine profiling for in-depth proteome-wide analysis and covalent ligand discovery. |
| Databáze: | OpenAIRE |
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