Abstract
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.
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Acknowledgments
We thank Maria-Eugenia Zaballa for her help in finalizing the manuscript. This work benefited from funding from the European Research Council under the European Union’s Seventh Framework Programme (FP/2007-2013)/ERC Grant Agreement no. 340260—PalmERa. This work was also supported by grants from the Swiss National Science Foundation (to G.v.d.G), and the Swiss SystemsX.ch initiative evaluated by the Swiss National Science Foundation (LipidX) (to G.v.d.G).
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Abrami, L., Denhardt-Eriksson, R.A., Hatzimanikatis, V., van der Goot, F.G. (2019). Dynamic Radiolabeling of S-Palmitoylated Proteins. In: Linder, M. (eds) Protein Lipidation. Methods in Molecular Biology, vol 2009. Humana, New York, NY. https://doi.org/10.1007/978-1-4939-9532-5_9
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DOI: https://doi.org/10.1007/978-1-4939-9532-5_9
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