Protein palmitoylation is a pivotal lipid modification that dictates membrane trafficking specificity. Here, we find phosphatidylinositol 4-kinase IIα (PI4KIIα) is the essential for transport of palmitoylated cargoes from the trans-Golgi network (TGN) to the plasma membrane (PM). Here, we performed quantitative stable isotope labeling by amino acids in cell culture (SILAC) proteomics in wild-type (WT) and PI4K2A-/- cells. Isolated PM fractions were divided for two parallel analyses: one was directly subjected to mass spectrometry, identifying PI4KIIα-dependent PM proteins; the other was first enriched for palmitoylated proteins via Acyl-RAC analysis, identifying PI4KIIα-dependent palmitoylated PM proteins.