Protocol for clickable photoaffinity labeling and quantitative chemical proteomics

Abstract

Summary: Here, we describe a protocol for a photoaffinity labeling probe strategy for target deconvolution in live cells. We made a chemical probe by incorporation of a photoreactive group to covalently cross-link with adjacent amino acid residues upon UV irradiation. Click chemistry-based enrichment captures labeled proteins for proteomic analysis. Here, we detail specifics for finding targets of LXRβ, but the protocol has potential for application to other targets.For complete details on the use and execution of this protocol, please refer to Seneviratne et al. (2020).

Keywords

• Cell-based Assays
• Molecular/Chemical Probes
• Protein Biochemistry
• Proteomics
• Mass Spectrometry