Dynamic SILAC to Determine Protein Turnover in Neurons and Glia

Methods Mol Biol. 2023:2603:1-17. doi: 10.1007/978-1-0716-2863-8_1.

Abstract

Cellular protein turnover-the net result of protein synthesis and degradation-is crucial to maintain protein homeostasis and cellular function under steady-state conditions and to enable cells to remodel their proteomes upon a perturbation. In brain cells, proteins are continuously turned over at different rates depending on various factors including cell type, subcellular localization, cellular environment, and neuronal activity. Here we describe a workflow for the analysis of protein synthesis, degradation, and turnover in primary cultured rat neurons and glia using dynamic/pulsed SILAC and mass spectrometry.

Keywords: Dynamic SILAC; Glia; Mass spectrometry; Neurons; Protein degradation; Protein synthesis; Protein turnover; SILAC.

MeSH terms

  • Animals
  • Isotope Labeling / methods
  • Mass Spectrometry
  • Neuroglia* / metabolism
  • Neurons / metabolism
  • Proteolysis
  • Proteome* / metabolism
  • Rats

Substances

  • Proteome