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Related Experiment Video

Updated: Jan 5, 2026

Identification of protein complexes with quantitative proteomics in S. cerevisiae
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Mapping the Saccharomyces cerevisiae Spatial Proteome with High Resolution Using hyperLOPIT.

Daniel J H Nightingale1,2, Stephen G Oliver2, Kathryn S Lilley3,4

  • 1Cambridge Centre for Proteomics, Department of Biochemistry, University of Cambridge, Cambridge, UK.

Methods in Molecular Biology (Clifton, N.J.)
|October 12, 2019
PubMed
Summary

This study introduces hyperplexed localization of organelle proteins by isotope tagging (hyperLOPIT) for high-throughput protein localization. This method maps the spatial proteome in yeast with high resolution and throughput.

Keywords:
OrganelleProtein localizationSaccharomyces cerevisiaeSpatial proteomicsSubcellular fractionationhyperLOPIT

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Area of Science:

  • Cell Biology
  • Proteomics
  • Biochemistry

Background:

  • Subcellular protein localization is crucial for cellular function.
  • Understanding organelle proteomes enhances knowledge of cellular homeostasis and dynamics.
  • High-throughput methods are needed for comprehensive spatial proteome mapping.

Purpose of the Study:

  • To present a protocol for unbiased, high-throughput study of protein subcellular localization.
  • To enable deep sampling and high-resolution mapping of the spatial proteome in Saccharomyces cerevisiae.
  • To detail the hyperplexed localization of organelle proteins by isotope tagging (hyperLOPIT) method.

Main Methods:

  • Biochemical fractionation of Saccharomyces cerevisiae.
  • High-resolution mass spectrometry-based protein quantitation.
  • Utilizing TMT 10-plex isobaric tags for hyperLOPIT.

Main Results:

  • Determination of subcellular localizations for thousands of proteins simultaneously.
  • High-throughput and high-resolution mapping of the spatial proteome.
  • Unbiased assessment of protein distribution within cellular compartments.

Conclusions:

  • The hyperLOPIT protocol provides a powerful tool for spatial proteome analysis.
  • This method significantly advances the study of cellular homeostasis and dynamics.
  • Enables parallel determination of protein localizations for deep proteome sampling.