Mapping the Saccharomyces cerevisiae Spatial Proteome with High Resolution Using hyperLOPIT.

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Cited by: 6

Abstract

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.

Keywords

Spatial Proteomics

Organelle

Protein localization

Saccharomyces cerevisiae

Spatial proteomics

Subcellular fractionation

hyperLOPIT

MeSH terms

Cell Fractionation

Mass Spectrometry

Proteome

Proteomics

Saccharomyces cerevisiae

Saccharomyces cerevisiae Proteins

Authors

Nightingale, Daniel J H

Oliver, Stephen G

Lilley, Kathryn S

Recommend literature

1. The subcellular organisation of Saccharomyces cerevisiae.

2. Dynamic Organellar Maps for Spatial Proteomics.

3. Organellar Maps Through Proteomic Profiling - A Conceptual Guide.

4. Using hyperLOPIT to perform high-resolution mapping of the spatial proteome.

5. A Protocol to Map the Spatial Proteome Using HyperLOPIT in Saccharomyces cerevisiae.

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