The human MDR3 P-glycoprotein promotes translocation of phosphatidylcholine through the plasma membrane of fibroblasts from transgenic mice.
The mouse mdr2 P-glycoprotein (P-gp) and its human MDR3 homologue are present in high concentrations in the canalicular membrane of hepatocytes. Mice lacking this protein are unable to secrete phosphatidylcholine (PC) into bile, suggesting that this P-gp is a PC translocator. We have tested this in fibroblasts from transgenic mice expressing the MDR3 gene under a vimentin promoter. Transgenic and control fibroblasts were incubated with [14C]choline to label PC. When the labeled cells were incubated with a PC transfer protein and acceptor liposomes, transfer of radioactive PC was enhanced in transgenic cells relative to the wild type controls. We conclude that the MDR3 P-glycoprotein is able to promote the transfer of PC from the inner to the outer leaflet of the plasma membrane, supporting the idea that this protein functions as a PC flippase.
Division of Molecular Biology, The Netherlands Cancer Institute, Amsterdam.
A J
AJ
Smith
J L
JL
Timmermans-Hereijgers
B
B
Roelofsen
K W
KW
Wirtz
W J
WJ
van Blitterswijk
J J
JJ
Smit
A H
AH
Schinkel
P
P
Borst
D020168:ATP Binding Cassette Transporter, Subfamily B, Member 1 / Q000235:genetics / Q000502:physiology*; D000818:Animals; D001692:Biological Transport; D002250:Carbon Radioisotopes; D002462:Cell Membrane / Q000378:metabolism*; D002794:Choline / Q000378:metabolism; D005347:Fibroblasts / Q000378:metabolism*; D007700:Kinetics; D008081:Liposomes / Q000378:metabolism; D051379:Mice; D008822:Mice, Transgenic; D010713:Phosphatidylcholines / Q000378:metabolism*; D011401:Promoter Regions, Genetic; D014746:Vimentin / Q000235:genetics
D020168:ATP Binding Cassette Transporter, Subfamily B, Member 1; D002250:Carbon Radioisotopes; D008081:Liposomes; D010713:Phosphatidylcholines; D014746:Vimentin; D002794:Choline