Affinity-Based Selectivity Profiling of an In-Class Selective Competitive Inhibitor of Acyl Protein Thioesterase 2
| Autor: | Sang Joon Won, Brandon T. Ruotolo, Sin Ye Hwang, Fei San Chong, Jaimeen D. Majmudar, Brent R. Martin, Joseph D. Eschweiler |
|---|---|
| Jazyk: | angličtina |
| Rok vydání: | 2016 |
| Předmět: |
chemistry.chemical_classification
010405 organic chemistry Kinase Metabolite Organic Chemistry Activity-based proteomics Serine hydrolase Biology 010402 general chemistry 01 natural sciences Biochemistry 0104 chemical sciences ACYL PROTEIN THIOESTERASE 2 chemistry.chemical_compound Enzyme chemistry Drug Discovery Selectivity Fluorescence anisotropy |
| Popis: | Activity-based protein profiling (ABPP) has revolutionized the discovery and optimization of active-site ligands across distinct enzyme families, providing a robust platform for in-class selectivity profiling. Nonetheless, this approach is less straightforward for profiling reversible inhibitors and does not access proteins outside the ABPP probe’s target profile. While the active-site competitive acyl protein thioesterase 2 inhibitor ML349 (Ki = 120 nM) is highly selective within the serine hydrolase enzyme family, it could still interact with other cellular targets. Here we present a chemoproteomic workflow to enrich and profile candidate ML349-binding proteins. In human cell lysates, biotinylated-ML349 enriches a recurring set of proteins, including metabolite kinases and flavin-dependent oxidoreductases that are potentially enhanced by avidity-driven multimeric interactions. Confirmatory assays by native mass spectrometry and fluorescence polarization quickly rank-ordered these weak off-targets, provi... |
| Databáze: | OpenAIRE |
| Externí odkaz: |
|