Isolation and Characterization of Lipid Particles from Yeast
Lipid particles (lipid droplets) are cell compartments destined for the storage of neutral lipids. These particles are present in all eukaryotic organisms as well as in few prokaryotes. Here, a procedure for lipid particle isolation from the budding yeast
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Introduction . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 4224
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Cell Growth . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 4225
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Preparation of Spheroplasts . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 4225
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Preparation of Lipid Particles . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 4225
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Note for Protein Analyses . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 4226
6 Solutions and Material . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 4226 6.1 Solutions and Specific Chemicals . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 4226 6.2 Special Equipment . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 4226 7
Troubleshooting . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 4227
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Research Needs . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 4227
K. N. Timmis (ed.), Handbook of Hydrocarbon and Lipid Microbiology, DOI 10.1007/978-3-540-77587-4_330, # Springer-Verlag Berlin Heidelberg, 2010
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Isolation and Characterization of Lipid Particles from Yeast
Abstract: Lipid particles (lipid droplets) are cell compartments destined for the storage of neutral lipids. These particles are present in all eukaryotic organisms as well as in few prokaryotes. Here, a procedure for lipid particle isolation from the budding yeast Saccharomyces cerevisiae is described. A simple protein pattern and huge amounts of neutral lipids are a characteristic of an isolated lipid particle fraction. Advices to overcome problems during lipid particle preparation are included.
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Introduction
Lipid particles (lipid droplets) are intracellular storage compartments for neutral lipids. These cell compartments are formed in all eukaryotic organisms, e.g., mammals, plants and fungi, as well as in few prokaryotes of the gram-positive genera (reviewed in Zweytick et al., 2000). Neutral lipids–mainly triacylglycerols and steryl esters–are forming the hydrophobic core of a lipid particle. This core is surrounded by a phospholipid monolayer with few characteristic proteins embedded (See also > Chapter 35, Vol. 1, Part 5). The neutral lipid composition of the hydrophobic core of lipid particles is highly dependent on the source of this compartment. As an example, lipid particles of adipocytes consist mainly of triacylglycerols, steryl esters are the main component of a particle of
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