Pulse SILAC Approaches to the Measurement of Cellular Dynamics

Justin Snider, Daifeng Wang, Daniel F. Bogenhagen, John D. Haley

Research output: Contribution to journalReview articlepeer-review

6 Scopus citations

Abstract

The global measurement of assembly and turnover of protein containing complexes within cells has advanced with the development of pulse stable isotope labelled amino acid approaches. Stable isotope labeling with amino acids in cell culture (SILAC) allows the incorporation of "light" 12-carbon amino acids or "heavy" 13-carbon amino acids into cells or organisms and the quantitation of proteins and peptides containing these amino acid tags using mass spectrometry. The use of these labels in pulse or pulse-chase scenarios has enabled measurements of macromolecular dynamics in cells, on time scales of several hours. Here we review advances with this approach and alternative or parallel strategies. We also examine the statistical considerations impacting datasets detailing mitochondrial assembly, to highlight key parameters in establishing significance and reproducibility.

Original languageEnglish (US)
Pages (from-to)575-583
Number of pages9
JournalAdvances in experimental medicine and biology
Volume1140
DOIs
StatePublished - 2019
Externally publishedYes

Keywords

  • Protein complex assembly
  • Protein synthesis
  • Pulse SILAC

ASJC Scopus subject areas

  • Biochemistry, Genetics and Molecular Biology(all)

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