title: The human multidrug resistance protein MRP1 translocates sphingolipid analogs across the plasma membrane.
abstract: Recently, we have provided evidence that the ABC-transporter MDR1 P-glycoprotein translocates analogs of various lipid classes across the apical plasma membrane of polarized LLC-PK1 cells transfected with MDR1 cDNA. Here, we show that expression of the basolateral ABC-transporter MRP1 (the multidrug resistance protein) induced lipid transport to the exoplasmic leaflet of the basolateral plasma membrane of LLC-PK1 cells at 15 degreesC. C6-NBD-glucosylceramide synthesized on the cytosolic side of the Golgi complex, but not C6-NBD-sphingomyelin synthesized in the Golgi lumen, became accessible to depletion by BSA in the basal culture medium. This suggests the absence of vesicular traffic and direct translocation of C6-NBD-glucosylceramide by MRP1 across the basolateral membrane. In line with this, transport of the lipid to the exoplasmic leaflet depended on the intracellular glutathione concentration and was inhibited by the MRP1-inhibitors sulfinpyrazone and indomethacin, but not by the MDR1 P-glycoprotein inhibitor PSC 833. In contrast to the broad substrate specificity of the MDR1 P-glycoprotein, MRP1 selectively transported C6-NBD-glucosylceramide and C6-NBD-sphingomyelin, the latter only when it was released from the Golgi lumen by brefeldin A. This shows the specific nature of the lipid translocation. We conclude that the transport activity of MDR1 P-glycoprotein and MRP1 must be taken into account in studies on the transport of lipids to the cell surface.
authors.affiliation: Department of Cell Biology and Histology, Academic Medical Center, University of Amsterdam, PO Box 22700, The Netherlands.
authors.forename: R J
authors.initials: RJ
authors.lastname: Raggers
authors.forename: A
authors.initials: A
authors.lastname: van Helvoort
authors.forename: R
authors.initials: R
authors.lastname: Evers
authors.forename: G
authors.initials: G
authors.lastname: van Meer
mesh_terms: D000818:Animals; D002462:Cell Membrane / Q000378:metabolism*; D002478:Cells, Cultured; D004268:DNA-Binding Proteins / Q000378:metabolism*; D005963:Glucosylceramides / Q000378:metabolism; D006801:Humans; D007668:Kidney / Q000378:metabolism; D050356:Lipid Metabolism; D027425:ATP-Binding Cassette, Sub-Family C Proteins; D000074081:MutS Homolog 3 Protein; D010069:Oxadiazoles / Q000378:metabolism; D010713:Phosphatidylcholines / Q000378:metabolism; D010742:Phospholipid Ethers / Q000378:metabolism; D013107:Sphingolipids / Q000378:metabolism*; D013379:Substrate Specificity; D013552:Swine; D014162:Transfection
chemical_list: D004268:DNA-Binding Proteins; D005963:Glucosylceramides; D027425:ATP-Binding Cassette, Sub-Family C Proteins; D000074081:MutS Homolog 3 Protein; D010069:Oxadiazoles; D010713:Phosphatidylcholines; D010742:Phospholipid Ethers; D013107:Sphingolipids; C095088:6-(N-(7-nitro-2,1,3-benzoxadiazol-4-yl)amino)hexanoyl-glucosylceramide; C078444:MSH3 protein, human; C026659:edelfosine; C044745:1-palmitoyl-2-(6-(N-(7-nitrobenz)-2-oxa-1,3-diazol-4-yl)aminocaproyl)phosphatidylcholine; C421109:multidrug resistance-associated protein 1