Assessing Budding Yeast Phosphoproteome Dynamics in a Time-Resolved Manner using TMT10plex Mass Tag Labeling

STAR Protoc. 2020 Jun 19;1(1):100022. doi: 10.1016/j.xpro.2020.100022.

Abstract

Amine-reactive Tandem Mass Tag 10plex (TMT10plex) labeling permits multiplexed protein identification and quantitative analysis by tandem mass spectrometry (MS/MS). We have used this technology to label 20 Saccharomyces cerevisiae samples collected in a time-resolved manner from a wild-type and phosphatase mutant background to characterize phosphoproteome dynamics. Here, we provide a detailed protocol for biological and mass spectrometry sample preparation and analysis. For complete details on the use and execution of this protocol, please refer to Touati et al. (2019).

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Mutation
  • Phosphoric Monoester Hydrolases / genetics
  • Proteome / analysis*
  • Saccharomyces cerevisiae / chemistry*
  • Saccharomyces cerevisiae / genetics
  • Saccharomyces cerevisiae Proteins / analysis*
  • Tandem Mass Spectrometry / methods*

Substances

  • Proteome
  • Saccharomyces cerevisiae Proteins
  • Phosphoric Monoester Hydrolases