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Open AccessArticle

Patches and Blebs: A Comparative Study of the Composition and Biophysical Properties of Two Plasma Membrane Preparations from CHO Cells

1
Instituto Biofisika (CSIC, UPV/EHU), Campus Universitario, 48940 Leioa, Spain
2
Departamento de Bioquímica, Universidad del País Vasco, 48940 Leioa, Spain
3
NCCR Chemical Biology, Department of Biochemistry, University of Geneva, 1211 Geneva, Switzerland
*
Author to whom correspondence should be addressed.
Int. J. Mol. Sci. 2020, 21(7), 2643; https://doi.org/10.3390/ijms21072643
Received: 1 April 2020 / Revised: 6 April 2020 / Accepted: 8 April 2020 / Published: 10 April 2020
(This article belongs to the Special Issue Lipid-Protein and Protein-Protein Interactions in Membranes)
This study was aimed at preparing and characterizing plasma membranes (PM) from Chinese Hamster Ovary (CHO) cells. Two methods of PM preparation were applied, one based on adhering cells to a poly-lysine-coated surface, followed by hypotonic lysis and removal of intracellular components, so that PM patches remain adhered to each other, and a second one consisting of bleb induction in cells, followed by separation of giant plasma membrane vesicles (GPMV). Both methods gave rise to PM in sufficient amounts to allow biophysical and biochemical characterization. Laurdan generalized polarization was used to measure molecular order in membranes, PM preparations were clearly more ordered than the average cell membranes (GP ≈0.450 vs. ≈0.20 respectively). Atomic force microscopy was used in the force spectroscopy mode to measure breakthrough forces of PM, both PM preparations provided values in the 4–6 nN range, while the corresponding value for whole cell lipid extracts was ≈2 nN. Lipidomic analysis of the PM preparations revealed that, as compared to the average cell membranes, PM were enriched in phospholipids containing 30–32 C atoms in their acyl chains but were relatively poor in those containing 34–40 C atoms. PM contained more saturated and less polyunsaturated fatty acids than the average cell membranes. Blebs (GPMV) and patches were very similar in their lipid composition, except that blebs contained four-fold the amount of cholesterol of patches (≈23 vs. ≈6 mol% total membrane lipids) while the average cell lipids contained 3 mol%. The differences in lipid composition are in agreement with the observed variations in physical properties between PM and whole cell membranes. View Full-Text
Keywords: giant plasma membrane vesicles; blebs; plasma membrane; general polarization; Atomic Force Microscopy (AFM); lipidomics giant plasma membrane vesicles; blebs; plasma membrane; general polarization; Atomic Force Microscopy (AFM); lipidomics
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MDPI and ACS Style

Monasterio, B.G.; Jiménez-Rojo, N.; García-Arribas, A.B.; Riezman, H.; Goñi, F.M.; Alonso, A. Patches and Blebs: A Comparative Study of the Composition and Biophysical Properties of Two Plasma Membrane Preparations from CHO Cells. Int. J. Mol. Sci. 2020, 21, 2643. https://doi.org/10.3390/ijms21072643

AMA Style

Monasterio BG, Jiménez-Rojo N, García-Arribas AB, Riezman H, Goñi FM, Alonso A. Patches and Blebs: A Comparative Study of the Composition and Biophysical Properties of Two Plasma Membrane Preparations from CHO Cells. International Journal of Molecular Sciences. 2020; 21(7):2643. https://doi.org/10.3390/ijms21072643

Chicago/Turabian Style

Monasterio, Bingen G.; Jiménez-Rojo, Noemi; García-Arribas, Aritz B.; Riezman, Howard; Goñi, Félix M.; Alonso, Alicia. 2020. "Patches and Blebs: A Comparative Study of the Composition and Biophysical Properties of Two Plasma Membrane Preparations from CHO Cells" Int. J. Mol. Sci. 21, no. 7: 2643. https://doi.org/10.3390/ijms21072643

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