Dynamic Radiolabeling of S-Palmitoylated Proteins

Proteins can be radiolabeled either during synthesis, typically using ³⁵S-cysteine/methionine (³⁵S-Cys/Met), or after synthesis, by adding a radiolabeled posttranslational modification. Here we describe how protein S-palmitoylation, and its dynamics, can be monitored by ³H-palmitate labeling and how the importance of S-palmitoylation in protein biogenesis and turnover can be investigated using ³⁵S-Cys/Met pulse-chase metabolic labeling. Proteins frequently have multiple palmitoylation sites. The importance thereof on the design and interpretation of metabolic labeling experiments is discussed.