Cysteine-reactive ligand discovery in proteomes
US10782295B2 · kind B2 · utility
Assignee
Inventors
Key dates
| Filing date | Aug 13, 2014 |
| Grant date | Sep 22, 2020 |
| Priority date | — |
| Expiry date | Dec 17, 2034 |
Classification
- Technology area (CPC G)Physics
- CPC primaryG01N2500/10
- WIPO fieldMeasurement
- WIPO sectorInstruments
Abstract
Cells produce electrophilic products with the potential to modify and affect the function of proteins. Chemoproteomic methods have provided a means to qualitatively inventory proteins targeted by endogenous electrophiles; however, ascertaining the potency and specificity of these reactions to identify the most sensitive sites in the proteome to electrophilic modification requires more quantitative methods. Here, we describe a competitive activity-based profiling method for quantifying the reactivity of electrophilic compounds against 1000+ cysteines in parallel in the human proteome. Using this approach, we identify a select set of proteins that constitute hot spots for modification by various lipid-derived electrophiles, including the oxidative stress product 4-hydroxnonenal (HNE). We show that one of these proteins, ZAK kinase, is labeled by HNE on a conserved, active site-proximal cysteine, resulting in enzyme inhibition to create a negative feedback mechanism that can suppress the activation of JNK pathways by oxidative stress.
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