ArticleYeast (Chichester, England)2014
A protocol for the subcellular fractionation of Saccharomyces cerevisiae using nitrogen cavitation and density gradient centrifugation.
Article in Yeast (Chichester, England), 2014. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 12 papers.
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Who cites it
12 citing papers in PubMed.
- Ribosomal protein Rps29/uS14 contributes to 18S rRNA maturation and its abundance regulates osmotic stress response in S. cerevisiae.Nucleic acids research · 2025Article
- Fine-tuning the yeast GAL10 promoter and growth conditions for efficient recombinant membrane protein production and purification.Protein science : a publication of the Protein Society · 2025Article
- Optimization of Cell Membrane Purification for the Preparation and Characterization of Cell Membrane Liposomes.Small methods · 2024Article
- Comprehensive Analysis of Proteasomal Complexes in Mouse Brain Regions Detects ENO2 as a Potential Partner of the Proteasome in the Striatum.Cellular and molecular neurobiology · 2022Article
- Review
- Investigating molecular crowding during cell division and hyperosmotic stress in budding yeast with FRET.Current topics in membranes · 2021Article
- A Comprehensive Subcellular Atlas of the Toxoplasma Proteome via hyperLOPIT Provides Spatial Context for Protein Functions.Cell host & microbe · 2020Article
- Mitochondrial cysteinyl-tRNA synthetase is expressed via alternative transcriptional initiation regulated by energy metabolism in yeast cells.The Journal of biological chemistry · 2019Article
- A Protocol to Map the Spatial Proteome Using HyperLOPIT inBio-protocol · 2019Article
- Mapping the Saccharomyces cerevisiae Spatial Proteome with High Resolution Using hyperLOPIT.Methods in molecular biology (Clifton, N.J.) · 2019Article
- Investigation of an optimal cell lysis method for the study of the zinc metalloproteome of Histoplasma capsulatum.Analytical and bioanalytical chemistry · 2017Article
- Using hyperLOPIT to perform high-resolution mapping of the spatial proteome.Nature protocols · 2017Article
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Authors and funding
3 authors.
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Abstract
Most protocols for yeast subcellular fractionation involve the use of mechanical shear forces to lyse the spheroplasts produced by the enzymatic digestion of the Saccharomyces cerevisiae cell wall. These mechanical homogenization procedures often involve the manual use of devices such as the Dounce homogenizer, and so are very operator-dependent and, in consequence, lack reproducibility. Here, we report a highly reproducible method of homogenizing yeast cells based on nitrogen cavitation. This has been optimized to allow efficient release of subcellular compartments that show a high degree of integrity. The protocol remains effective and reproducible across a range of sample volumes and buffer environments. The subsequent separation method, which employs both sucrose and iodixanol density gradients, has been developed to resolve the major membrane-bound compartments of S. cerevisiae. We present an integrated protocol that is fast, facile, robust and efficient and that will enable 'omics' studies of the subcellular compartments of S. cerevisiae and other yeasts.
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Read under generation 80e0d062 · epoch 390. Bibliography from PubMed, PubMed Central and OpenAlex; grants from NIH RePORTER; trial links from ClinicalTrials.gov; estimates, votes and beliefs from the OpenQuestion graph.