Proton NMR visible mobile lipid signals in sensitive and multidrug-resistant K562 cells are modulated by rafts

BACKGROUND: Most cancer cells are characterized by mobile lipids visible on proton NMR ((1)H-NMR), these being comprised mainly of methyl and methylene signals from lipid acyl chains. Erythroleukemia K562 cells show narrow signals at 1.3 and 0.9 ppm, corresponding to mobile lipids (methylene and methyl, respectively), which are reduced when K562 cells are multidrug resistant (MDR). While the significance of the mobile lipids is unknown, their subcellular localization is still a matter of debate and may lie in the membrane or the cytoplasm. In this study, we investigate the role of cholesterol in the generation of mobile lipid signals. RESULTS: The proportion of esterified cholesterol was found to be higher in K562-sensitive cells than in resistant cells, while the total cholesterol content was identical in both cell lines. Cholesterol extraction in the K562 wild type (K562wt) cell line and its MDR counterpart (K562adr), using methyl-β-cyclodextrin, was accompanied by a rise of mobile lipids in K562wt cells only. The absence of caveolae was checked by searching for the caveolin-1 protein in K562wt and K562adr cells. However, cholesterol was enriched in another membrane microdomain designated as "detergent-insoluble glycosphingomyelin complexes" or rafts. These microdomains were studied after extraction with triton X-100, a mild non-ionic detergent, revealing mobile lipid signals preserved only in the K562wt spectra. Moreover, following perturbation/disruption of these microdomains using sphingomyelinase, mobile lipids increased only in K562wt cells. CONCLUSION: These results suggest that cholesterol and sphingomyelin are involved in mobile lipid generation via microdomains of detergent-insoluble glycosphingomyelin complexes such as rafts. Increasing our knowledge of membrane microdomains in sensitive and resistant cell lines may open up new possibilities in resistance reversion

beta-Dystroglycan can be revealed in microsomes from mdx mouse muscle by detergent treatment

International audiencebeta-Dystroglycan is the central member of a transmembrane protein complex of the skeletal muscle plasma membrane. Since it was not detected in dystrophin-deficient skeletal muscles, a disruption of the complex was thought to be involved in the dystrophic process. We report here that beta-dystroglycan is actually present at normal levels in mdx mouse muscle plasma membrane: treatment with cholate detergent is able to reveal its presence by SDS-PAGE and immunoblotting. This result shows that, in dystrophin-deficient muscles, beta-dystroglycan is indeed targeted to the plasma membrane but remains inaccessible to classical solubilizing treatments and to antibodies used for immunolocalizatio

L’autorégulation des apprentissages dans une formation pour adulte. L’exemple de la demande d’aide

International audienc

L’autorégulation des apprentissages dans une formation pour adulte. L’exemple de la demande d’aide

International audienc

L’autorégulation des apprentissages dans une formation pour adulte. L’exemple de la demande d’aide

International audienc