Mapping the Saccharomyces cerevisiae Spatial Proteome with High Resolution Using hyperLOPIT.
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The subcellular localization of proteins is a posttranslational modification of paramount importance. The ability to study subcellular and organelle proteomes improves our understanding of cellular homeostasis and cellular dynamics. In this chapter, we describe a protocol for the unbiased and high-throughput study of protein subcellular localization in the yeast Saccharomyces cerevisiae: hyperplexed localization of organelle proteins by isotope tagging (hyperLOPIT), which involves biochemical fractionation of Saccharomyces cerevisiae and high resolution mass spectrometry-based protein quantitation using TMT 10-plex isobaric tags. This protocol enables the determination of the subcellular localizations of thousands of proteins in parallel in a single experiment and thereby deep sampling and high-resolution mapping of the spatial proteome.


Spatial Proteomics
Protein localization
Saccharomyces cerevisiae
Spatial proteomics
Subcellular fractionation

MeSH terms

Cell Fractionation
Mass Spectrometry
Saccharomyces cerevisiae
Saccharomyces cerevisiae Proteins


Nightingale, Daniel J H
Oliver, Stephen G
Lilley, Kathryn S

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