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Modeling the effects of cyclodextrin on intracellular membrane vesicles from Cos-7 cells prepared by sonication and carbonate treatment
- Source :
- PeerJ, Vol 3, p e1351 (2015), PeerJ
- Publication Year :
- 2015
- Publisher :
- PeerJ Inc., 2015.
-
Abstract
- Cholesterol has important functions in the organization of membrane structure and this may be mediated via the formation of cholesterol-rich, liquid-ordered membrane microdomains often referred to as lipid rafts. Methyl-beta-cyclodextrin (cyclodextrin) is commonly used in cell biology studies to extract cholesterol and therefore disrupt lipid rafts. However, in this study we reassessed this experimental strategy and investigated the effects of cyclodextrin on the physical properties of sonicated and carbonate-treated intracellular membrane vesicles isolated from Cos-7 fibroblasts. We treated these membranes, which mainly originate from thetrans-Golgi network and endosomes, with cyclodextrin and measured the effects on their equilibrium buoyant density, protein content, represented by the palmitoylated protein phosphatidylinositol 4-kinase type IIα, and cholesterol. Despite the reduction in mass stemming from cholesterol removal, the vesicles became denser, indicating a possible large volumetric decrease, and this was confirmed by measurements of hydrodynamic vesicle size. Subsequent mathematical analyses demonstrated that only half of this change in membrane size was attributable to cholesterol loss. Hence, the non-selective desorption properties of cyclodextrin are also involved in membrane size and density changes. These findings may have implications for preceding studies that interpreted cyclodextrin-induced changes to membrane biochemistry in the context of lipid raft disruption without taking into account our finding that cyclodextrin treatment also reduces membrane size.
- Subjects :
- Endosome
Biophysics
lcsh:Medicine
General Biochemistry, Genetics and Molecular Biology
PI 4-kinase
chemistry.chemical_compound
Cyclodextrin
Mathematical Biology
Lipid raft
chemistry.chemical_classification
Cholesterol
General Neuroscience
Vesicle
Endoplasmic reticulum
lcsh:R
Membrane structure
Membrane
Cell Biology
General Medicine
Biochemistry
chemistry
TGN
lipids (amino acids, peptides, and proteins)
General Agricultural and Biological Sciences
Subjects
Details
- Language :
- English
- ISSN :
- 21678359
- Volume :
- 3
- Database :
- OpenAIRE
- Journal :
- PeerJ
- Accession number :
- edsair.doi.dedup.....58435bb30c1836cc6ec5f15b6556f74f