Measurement and utilization of the proteomic reactivity by mass spectrometry

Abstract Chemical proteomics, which involves studying the covalent modifications of proteins by small molecules, has significantly contributed to our understanding of protein function and has become an essential tool in drug discovery. Mass spectrometry (MS) is the primary method for identifying and quantifying protein‐small molecule adducts. In this review, we discuss various methods for measuring proteomic reactivity using MS and covalent proteomics probes that engage through reactivity‐driven and proximity‐driven mechanisms. We highlight the applications of these methods and probes in live‐cell measurements, drug target identification and validation, and characterizing protein‐small molecule interactions. We conclude the review with current developments and future opportunities in the field, providing our perspectives on analytical considerations for MS‐based analysis of the proteomic reactivity landscape.