Palmitoylation, the attachment of fatty acids (predominantly palmitate) onto cellular proteins, has emerged as a widespread post-translational modification of both transmembrane and peripheral membrane proteins. Recent breakthroughs in the palmitoylation field came with the discovery of the “DHHC” protein family that collectively regulate cellular palmitoylation dynamics; at least 24 of these DHHC proteins are encoded in the human genome. I will discuss how the interaction of specific DHHC isoforms with peripheral membrane protein substrates is encoded. In addition, I will discuss how multiple palmitoylation of individual peripheral membrane proteins affects their intracellular trafficking and dynamics in normal and disease states.