Localization and regulation of SR-BI in membrane rafts of HepG2 cells

D Rhainds, P Bourgeois, G Bourret… - Journal of Cell …, 2004 - journals.biologists.com
D Rhainds, P Bourgeois, G Bourret, K Huard, L Falstrault, L Brissette
Journal of Cell Science, 2004journals.biologists.com
The scavenger receptor class B, type I (SR-BI) mediates cholesteryl esters (CE) selective
uptake from low density lipoprotein (LDL) and high-density lipoprotein (HDL) particles. In a
number of tissues expressing caveolin, SR-BI is localized in caveolae. We show using
detergent-free sucrose gradients that SR-BI is found in membrane rafts devoid of caveolin-1
in the human hepatoma HepG2 cell. Perturbation of the structure of HepG2 cell membrane
rafts with cholesterol oxidase or sphingomyelinase decreased LDL-CE association due to …
The scavenger receptor class B, type I (SR-BI) mediates cholesteryl esters (CE) selective uptake from low density lipoprotein (LDL) and high-density lipoprotein (HDL) particles. In a number of tissues expressing caveolin, SR-BI is localized in caveolae. We show using detergent-free sucrose gradients that SR-BI is found in membrane rafts devoid of caveolin-1 in the human hepatoma HepG2 cell. Perturbation of the structure of HepG2 cell membrane rafts with cholesterol oxidase or sphingomyelinase decreased LDL-CE association due to selective uptake by 60%, while HDL3-CE selective uptake was increased 2.3-fold by cholesterol oxidase but was not affected by sphingomyelinase. Sequestration of membrane cholesterol with filipin III decreased LDL-CE selective uptake by 25%, while it had no effect on HDL3-CE selective uptake. Extraction of cell membrane cholesterol with β-cyclodextrin increased LDL- and HDL3-CE selective uptake by 1.6-fold and 3-fold, respectively. We found that CE-selective uptake from both HDL and LDL occurs by a pathway involving retro-endocytosis in HepG2 cells. An analysis of the effect of SR-BI level on the expression of critical lipid sensor and lipid binding proteins was conducted with stable transformants of HepG2 cell overexpressing SR-BI. We found that liver-type fatty acid binding protein expression level is higher in SR-BI-overexpressing cells and that caveolin-1 and sterol response element binding protein-2 levels are reduced. Thus, in this hepatic cell model, SR-BI is associated with membrane rafts devoid of caveolin and its expression affects intracellular lipid binding and lipid sensor proteins. SR-BI-dependent LDL- and HDL-CE selective uptake are affected differently by the integrity of membrane rafts, but both occur by a retroendocytic pathway in HepG2 cells.
journals.biologists.com