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Type: Artigo de periódico
Title: Resistance Of Human Erythrocyte Membranes To Triton X-100 And C12e8.
Author: Crepaldi Domingues, Cleyton
Ciana, Annarita
Buttafava, Armando
Balduini, Cesare
de Paula, Eneida
Minetti, Giampaolo
Abstract: Lipid rafts are microdomains enriched in cholesterol and sphingolipids that contain specific membrane proteins. The resistance of domains to extraction by nonionic detergents at 4 degrees C is the commonly used method to characterize these structures that are operationally defined as detergent-resistant membranes (DRMs). Because the selectivity of different detergents in defining membrane rafts has been questioned, we have compared DRMs from human erythrocytes prepared with two detergents: Triton X-100 and C12E8. The DRMs obtained presented a cholesterol/protein mass ratio three times higher than in the whole membrane. Flotillin-2 was revealed in trace amounts in DRMs obtained with C12E8, but it was almost completely confined within the DRM fraction with Triton X-100. Differently, stomatin was found distributed in DRM and non-DRM fractions for both detergents. We have also measured the order parameter (S) of nitroxide spin labels inserted into DRMs by means of electron paramagnetic resonance. The 5- and 16-stearic acid spin label revealed significantly higher S values for DRMs obtained with either Triton X-100 or C12E8 in comparison to intact cells, while the difference in the S values between Triton X-100 and C12E8 DRMs was not statistically significant. Our results suggest that although the acyl chain packing is similar in DRMs prepared with either Triton X-100 or C12E8 detergent, protein content is dissimilar, with flotillin-2 being selectively enriched in Triton X-100 DRMs.
Subject: Blotting, Western
Electron Spin Resonance Spectroscopy
Electrophoresis, Polyacrylamide Gel
Erythrocyte Membrane
Membrane Microdomains
Membrane Proteins
Citation: The Journal Of Membrane Biology. v. 227, n. 1, p. 39-48, 2009-Jan.
Rights: fechado
Identifier DOI: 10.1007/s00232-008-9142-4
Date Issue: 2009
Appears in Collections:Unicamp - Artigos e Outros Documentos

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