N-myristoylation is a ubiquitous class of protein lipidation across eukaryotes and N-myristoyl transferase (NMT) has been proposed as an attractive drug target in several pathogens. Here we describe the first global chemoproteomic analysis of protein myristoylation in Toxoplasma gondii. Through quantitative mass spectrometry coupled with validated chemoproteomic tools (cleavable capture reagents (Broncel et al., 2015, Speers and Cravatt, 2005) and NMT inhibitor (Schlott et al., 2019)) that allow for experimental validation, we confidently identified 65 myristoylated proteins. This dataset represents a large fraction of the parasite’s myristoylated proteome and a prerequisite to investigate this modification in Toxoplasma.