Updated project metadata. Chemoproteomics is a key technology to characterize the mode of action of drugs, as it directly identifies the protein targets of bioactive compounds and aids in developing optimized leads. We have developed a drug target deconvolution approach with an automated data analysis pipeline, based on limited proteolysis coupled with mass spectrometry that works across species including in human cells (LiP-Quant). Here we demonstrate drug target identification by LiP-Quant across compound classes, including with drugs targeting kinases and phosphatases . We demonstrate that LiP-Quant identifies drug binding sites, a unique feature of this approach, and that it can be used to estimate drug EC50s in whole cell lysates . LiP-Quant identifies targets of both selective and promiscuous drugs and correctly discriminates drug binding to homologous proteins. We finally show that the LiP-Quant technology identifies targets of a novel research compound of biotechnological interest.