Cholesteryl ester is transported from caveolae to internal membranes as part of a caveolin-annexin II lipid-protein complex.
We previously demonstrated that in Chinese hamster ovary cells scavenger receptor, class B, type I-dependent selective cholesteryl ester uptake occurs in caveolae. In the present study we hypothesized that cholesteryl ester is transported from caveolae through the cytosol to an internal membrane by a caveolin chaperone complex similar to the one we originally described for the transport of newly synthesized cholesterol. To test this hypothesis we incubated Chinese hamster ovary cells expressing scavenger receptor, class B, type I with [(3)H]cholesteryl ester-labeled high density lipoprotein, subfractionated the cells and looked for a cytosolic pool of [(3)H]cholesteryl ester. The radiolabeled sterol initially appeared in the caveolae fraction, then in the cytosol, and finally in the internal membrane fraction. Caveolin IgG precipitated all of the [(3)H]cholesteryl ester associated with the cytosol. Co-immunoprecipitation studies demonstrated that in the presence of high density lipoprotein, but not low density lipoprotein or lipoprotein-deficient serum, caveolin IgG precipitated four proteins: annexin II, cyclophilin 40, caveolin, and cyclophilin A. Caveolin acylation-deficient mutants were used to demonstrate that acylation of cysteine 133 but not cysteine 143 or 156 is required for annexin II association with caveolin and the rapid transport of cholesteryl esters out of caveolae. We conclude that a caveolin-annexin II lipid-protein complex facilitates the rapid internalization of cholesteryl esters from caveolae.
Uittenbogaard
Annette
A
Department of Physiology, University of Kentucky Medical School, Lexington, Kentucky 40536, USA.
Everson
William V
WV
Matveev
Sergey V
SV
Smart
Eric J
EJ
D000818:Animals; D017306:Annexin A2 / Q000378:metabolism*; D001692:Biological Transport; D016466:CHO Cells; D021941:Caveolae / Q000378:metabolism; D051242:Caveolin 1; D022461:Caveolins / Q000378:metabolism*; D002460:Cell Line; D002462:Cell Membrane / Q000378:metabolism*; D002788:Cholesterol Esters / Q000378:metabolism*; D006224:Cricetinae; D003545:Cysteine / Q000737:chemistry; D003600:Cytosol / Q000378:metabolism; D004591:Electrophoresis, Polyacrylamide Gel; D008401:Gas Chromatography-Mass Spectrometry; D006801:Humans; D015151:Immunoblotting; D007074:Immunoglobulin G / Q000378:metabolism; D050356:Lipid Metabolism; D008077:Lipoproteins, LDL / Q000378:metabolism; D051379:Mice; D009154:Mutation; D019308:Palmitic Acid / Q000378:metabolism; D011233:Precipitin Tests; D011485:Protein Binding; D016622:Silver Staining; D013696:Temperature; D013997:Time Factors; D014162:Transfection
D017306:Annexin A2; C495540:CAV1 protein, human; C495541:Cav1 protein, mouse; D051242:Caveolin 1; D022461:Caveolins; D002788:Cholesterol Esters; D007074:Immunoglobulin G; D008077:Lipoproteins, LDL; D019308:Palmitic Acid; D003545:Cysteine