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.
CITATION STYLE
Nightingale, D. J. H., Oliver, S. G., & Lilley, K. S. (2019). Mapping the Saccharomyces cerevisiae Spatial Proteome with High Resolution Using hyperLOPIT. In Methods in Molecular Biology (Vol. 2049, pp. 165–190). Humana Press Inc. https://doi.org/10.1007/978-1-4939-9736-7_10
Mendeley helps you to discover research relevant for your work.