Identification of Protein Targets of Bioactive Small Molecules Using Randomly Photomodified Probes
Language English Country United States Media print-electronic
Document type Journal Article, Research Support, Non-U.S. Gov't
- MeSH
- Affinity Labels chemical synthesis chemistry radiation effects MeSH
- Aspartic Acid Endopeptidases antagonists & inhibitors chemistry MeSH
- Biotin chemistry MeSH
- Diazomethane analogs & derivatives chemical synthesis radiation effects MeSH
- Endopeptidases MeSH
- Fluoresceins chemistry MeSH
- Fluorescent Dyes chemistry MeSH
- Glutamate Carboxypeptidase II antagonists & inhibitors chemistry MeSH
- Mass Spectrometry methods MeSH
- Enzyme Inhibitors chemical synthesis chemistry radiation effects MeSH
- Microscopy, Confocal methods MeSH
- Polymethacrylic Acids chemistry MeSH
- Humans MeSH
- Membrane Proteins antagonists & inhibitors chemistry MeSH
- Cell Line, Tumor MeSH
- Proteomics methods MeSH
- Serine Endopeptidases chemistry MeSH
- Ultraviolet Rays MeSH
- Gelatinases antagonists & inhibitors chemistry MeSH
- Check Tag
- Humans MeSH
- Publication type
- Journal Article MeSH
- Research Support, Non-U.S. Gov't MeSH
- Names of Substances
- Affinity Labels MeSH
- Aspartic Acid Endopeptidases MeSH
- Biotin MeSH
- Diazomethane MeSH
- Duxon MeSH Browser
- Endopeptidases MeSH
- fibroblast activation protein alpha MeSH Browser
- Fluoresceins MeSH
- Fluorescent Dyes MeSH
- Glutamate Carboxypeptidase II MeSH
- Enzyme Inhibitors MeSH
- Polymethacrylic Acids MeSH
- Membrane Proteins MeSH
- Serine Endopeptidases MeSH
- Gelatinases MeSH
Identifying protein targets of bioactive small molecules often requires complex, lengthy development of affinity probes. We present a method for stochastic modification of small molecules of interest with a photoactivatable phenyldiazirine linker. The resulting isomeric mixture is conjugated to a hydrophilic copolymer decorated with biotin and a fluorophore. We validated this approach using known inhibitors of several medicinally relevant enzymes. At least a portion of the stochastic derivatives retained their binding to the target, enabling target visualization, isolation, and identification. Moreover, the mix of stochastic probes could be separated into fractions and tested for binding affinity. The structure of the active probe could be determined and the probe resynthesized to improve binding efficiency. Our approach can thus enable rapid target isolation, identification, and visualization, while providing information required for subsequent synthesis of an optimized probe.
References provided by Crossref.org
Polymer-Tethered Quenched Fluorescent Probes for Enhanced Imaging of Tumor-Associated Proteases
Polymer-tethered quenched fluorescent probes for enhanced imaging of tumor associated proteases
