Quantitative proteomics for Xenopus embryos II, data analysis

Matthew Sonnett, Meera Gupta, Thao Nguyen, Martin Wühr

Research output: Chapter in Book/Report/Conference proceedingChapter

4 Scopus citations

Abstract

The oocytes, embryos, and cell-free lysates of the frog Xenopus laevis have emerged as powerful models for quantitative proteomic experiments. In the accompanying paper (Chapter 13) we describe how to prepare samples and acquire multiplexed proteomics spectra from those. As an illustrative example we use a 10-stage developmental time series from the egg to stage 35 (just before hatching). Here, we outline how to convert the ~700,000 acquired mass spectra from this time series into protein expression dynamics for ~9000 proteins. We first outline a preliminary quality-control analysis to discover any errors that occurred during sample preparation. We discuss how peptide and protein identification error rates are controlled, and how peptide and protein species are quantified. Our analysis relies on the freely available MaxQuant proteomics pipeline. Finally, we demonstrate how to start interpreting this large dataset by clustering and gene-set enrichment analysis.

Original languageEnglish (US)
Title of host publicationMethods in Molecular Biology
PublisherHumana Press Inc.
Pages195-215
Number of pages21
DOIs
StatePublished - 2018

Publication series

NameMethods in Molecular Biology
Volume1865
ISSN (Print)1064-3745

All Science Journal Classification (ASJC) codes

  • Molecular Biology
  • Genetics

Keywords

  • Development
  • False discovery rate
  • Gene symbols
  • Gene-set enrichment
  • Mass spectrum
  • MaxQuant
  • Quantitative multiplexed proteomics
  • TMT
  • Xenopus laevis
  • k-means clustering

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