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SALVADOR, Dimitri; GLAVIER, Marie; TAVEAU, Jean-Christophe; LECOMTE, Sophie; LAMBERT, Olivier;
Minimal nanodisc without exogenous lipids for stabilizing membrane proteins in detergent-free buffer
Biochimica et biophysica acta. Biomembranes, 2019, 1861, 852-860
Membrane protein stabilization after detergent solubilization presents drawbacks for structural and biophysical studies, in particular that of a reduced stability in detergent micelles. Therefore, alternative methods are required for efficient stabilization. Lipid nanodisc made with the membrane scaffold protein MSP is a valuable system but requires a fine optimization of the lipid to protein ratio. We present here the use of the scaffold protein MSP without added lipids as a minimal system to stabilize membrane proteins. We show that this method is applicable to alpha-helical and beta-strands transmembrane proteins. This method allowed cryo-electron microscopy structural study of the bacterial transporter MexB. A protein quantification indicates that MexB is stabilized by two MSP proteins. This simplified and efficient method proposes a new advance in harnessing the MSP potential to stabilize membrane proteins.
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GERBOD-GIANNONE, Marie-Christine; DALLET, Laurence; NAUDIN, Grégoire; SAHIN, Annelise; POUSSARD, Sylvie; LAMBERT, Olivier;
Involvement of caveolin-1 and CD36 in native LDL endocytosis by endothelial cells
Biochimica et biophysica acta. General subjects, 2019, 1863, 830-838
Atherosclerosis is a lipid disease characterized by accumulation of low density lipoprotein (LDL) in the artery wall. The transport of LDL across the endothelium of coronary artery is an initiating event of atherosclerosis, whose mechanism remains poorly understood. In the last decade, it has been shown that in caveolin-1 (Cav-1) deficient mice, LDL infiltration in aorta wall is decreased and CD36 expression in aortas is down-regulated, leading to regression of atherosclerotic lesions. In the present study, we show that native LDL endocytosis is decreased in endothelial cells deficient in Cav-1 or CD36. We demonstrate that Cav-1 and CD36 interact in caveolae-rich domains by different biochemical approaches. In addition, confocal microscopy reveals some colocalization of Cav-1 with CD36. These findings indicate that caveolae and CD36 are involved in native LDL endocytosis and suggest that CD36 might be a good candidate for the transport of native LDL across the endothelium, an early event in atherosclerosis.
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