MDR3 P-glycoprotein, a phosphatidylcholine translocase, transports several cytotoxic drugs and directly interacts with drugs as judged by interference with nucleotide trapping.
The human MDR3 gene is a member of the multidrug resistance (MDR) gene family. The MDR3 P-glycoprotein is a transmembrane protein that translocates phosphatidylcholine. The MDR1 P-glycoprotein related transports cytotoxic drugs. Its overexpression can make cells resistant to a variety of drugs. Attempts to show that MDR3 P-glycoprotein can cause MDR have been unsuccessful thus far. Here, we report an increased directional transport of several MDR1 P-glycoprotein substrates, such as digoxin, paclitaxel, and vinblastine, through polarized monolayers of MDR3-transfected cells. Transport of other good MDR1 P-glycoprotein substrates, including cyclosporin A and dexamethasone, was not detectably increased. MDR3 P-glycoprotein-dependent transport of a short-chain phosphatidylcholine analog and drugs was inhibited by several MDR reversal agents and other drugs, indicating an interaction between these compounds and MDR3 P-gp. Insect cell membranes from Sf9 cells overexpressing MDR3 showed specific MgATP binding and a vanadate-dependent, N-ethylmaleimide-sensitive nucleotide trapping activity, visualized by covalent binding with [alpha-(32)P]8-azido-ATP. Nucleotide trapping was (nearly) abolished by paclitaxel, vinblastine, and the MDR reversal agents verapamil, cyclosporin A, and PSC 833. We conclude that MDR3 P-glycoprotein can bind and transport a subset of MDR1 P-glycoprotein substrates. The rate of MDR3 P-glycoprotein-mediated transport is low for most drugs, explaining why this protein is not detectably involved in multidrug resistance. It remains possible, however, that drug binding to MDR3 P-glycoprotein could adversely affect phospholipid or toxin secretion under conditions of stress (e.g. in pregnant heterozygotes with one MDR3 null allele).
Division of Molecular Biology and Center for Biomedical Genetics, The Netherlands Cancer Institute, Plesmanlaan 121, 1066 CX Amsterdam, The Netherlands.
A J
AJ
Smith
A
A
van Helvoort
G
G
van Meer
K
K
Szabo
E
E
Welker
G
G
Szakacs
A
A
Varadi
B
B
Sarkadi
P
P
Borst
D018435:ATP Binding Cassette Transporter, Subfamily B / Q000235:genetics / Q000378:metabolism*; D018528:ATP-Binding Cassette Transporters / Q000235:genetics / Q000378:metabolism*; D000255:Adenosine Triphosphate / Q000378:metabolism*; D000818:Animals; D001692:Biological Transport / Q000187:drug effects; D016764:Cell Polarity; D002478:Cells, Cultured; D016572:Cyclosporine / Q000494:pharmacology; D003524:Cyclosporins / Q000494:pharmacology; D003603:Cytotoxins / Q000378:metabolism*; D003630:Daunorubicin / Q000378:metabolism; D004077:Digoxin / Q000378:metabolism; D004847:Epithelial Cells / Q000166:cytology; D006801:Humans; D007559:Ivermectin / Q000378:metabolism; D007668:Kidney / Q000166:cytology; D017239:Paclitaxel / Q000378:metabolism; D004364:Pharmaceutical Preparations / Q000378:metabolism*; D010713:Phosphatidylcholines / Q000378:metabolism*; D011994:Recombinant Proteins / Q000378:metabolism; D018411:Spodoptera / Q000166:cytology; D013552:Swine; D014700:Verapamil / Q000494:pharmacology; D014747:Vinblastine / Q000378:metabolism
D018435:ATP Binding Cassette Transporter, Subfamily B; D018528:ATP-Binding Cassette Transporters; D003524:Cyclosporins; D003603:Cytotoxins; D004364:Pharmaceutical Preparations; D010713:Phosphatidylcholines; D011994:Recombinant Proteins; D014747:Vinblastine; D007559:Ivermectin; D004077:Digoxin; D016572:Cyclosporine; D000255:Adenosine Triphosphate; C095122:multidrug resistance protein 3; D014700:Verapamil; D017239:Paclitaxel; C070272:valspodar; D003630:Daunorubicin