| Literature DB >> 31602611 |
Daniel J H Nightingale1,2, Stephen G Oliver2, Kathryn S Lilley3,4.
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.Entities:
Keywords: Organelle; Protein localization; Saccharomyces cerevisiae; Spatial proteomics; Subcellular fractionation; hyperLOPIT
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Year: 2019 PMID： 31602611 DOI： 10.1007/978-1-4939-9736-7_10
Source DB: PubMed Journal: Methods Mol Biol ISSN： 1064-3745