| Article ID | Journal | Published Year | Pages | File Type |
|---|---|---|---|---|
| 1947534 | Biochimica et Biophysica Acta (BBA) - General Subjects | 2014 | 8 Pages |
•ATR-FTIR allowed monitoring changes in DRM composition after the cell treatment with DHA.•Multidimensional scaling allowed disclosing the components responsible of the observed spectral variance.•A rearrangement of membrane lipid acyl chains was observed after the cell treatment with DHA.•A decrease of sphingomyelin and cholesterol was found after DHA incorporation.
BackgroundDetergent resistant membranes (DRMs) are a useful model system for the in vitro characterization of cell membrane domains. Indeed, DRMs provide a simple model to study the mechanisms underlying several key cell processes based on the interplay between specific cell membrane domains on one hand, and specific proteins and/or lipids on the other. Considering therefore their biological relevance, the development of methods capable to provide information on the composition and structure of membrane domains and to detect their modifications is highly desirable. In particular, Fourier transform infrared (FTIR) spectroscopy is a vibrational tool widely used for the study not only of isolated and purified biomolecules but also of complex biological systems, including intact cells and tissues. One of the main advantages of this non-invasive approach is that it allows obtaining a molecular fingerprint of the sample under investigation in a rapid and label-free way.MethodsHere we present an FTIR characterization of DRM fractions purified from the human breast cancer cells MCF-7, before and after treatment with the omega 3 fatty acid docosahexaenoic acid (DHA), which was found to promote membrane microdomain reorganization.Results and ConclusionsWe will show that FTIR spectroscopy coupled with multivariate analysis enables to monitor changes in the composition of DRMs, induced in particular by the incorporation of DHA in cell membrane phospholipids.General significanceThis study paves the way for a new label-free characterization of specific membrane domains within intact cells, which could provide complementary information to the fluorescence approaches presently used.
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