Dynamic Radiolabeling of S-Palmitoylated Proteins

Laurence Abrami; Robin A Denhardt-Eriksson; Vassily Hatzimanikatis; F Gisou van der Goot

doi:10.1007/978-1-4939-9532-5_9

Dynamic Radiolabeling of S-Palmitoylated Proteins

Methods Mol Biol. 2019:2009:111-127. doi: 10.1007/978-1-4939-9532-5_9.

Authors

Laurence Abrami¹, Robin A Denhardt-Eriksson², Vassily Hatzimanikatis³, F Gisou van der Goot⁴

Affiliations

¹ Global Health Institute, School of Life Sciences, EPFL, Lausanne, Switzerland.
² Global Health Institute, Laboratory of Computational Systems Biotechnology, EPFL, Lausanne, Switzerland.
³ Laboratory of Computational Systems Biotechnology, EPFL, Lausanne, Switzerland.
⁴ Global Health Institute, School of Life Sciences, EPFL, Lausanne, Switzerland. [email protected].

PMID: 31152399
DOI: 10.1007/978-1-4939-9532-5_9

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.

Keywords: Data-driven mathematical modeling; Metabolic labeling; Protein S-acyltransferase (PAT); Pulse–chase; Radiolabeling; S-palmitoylation; Turnover.

Publication types

Research Support, Non-U.S. Gov't

MeSH terms

Cell Line, Transformed
Humans
Isotope Labeling / methods*
Lipoylation*
Palmitic Acid / metabolism*
Protein Processing, Post-Translational*
Sulfur Radioisotopes*

Substances

Sulfur Radioisotopes
Palmitic Acid