Expression and characterization of the multidrug resistance-associated protein in insect cells infected with a recombinant baculovirus.
The protein encoded by the multidrug resistance-associated protein (MRP) gene was examined after infection of SF21 insect cells with recombinant baculovirus containing a full-length MRP cDNA. The time course of appearance of the protein as determined by western blot analysis revealed that maximum levels occurred 2 days postinfection. The amount of MRP made in this system was somewhat variable, but levels that were about 4-fold greater than that found in HL60/ADR cells could be achieved. The protein appeared to be full-length but was present in a highly deglycosylated form. The P170 (MRP) was phosphorylated and located exclusively in membranes of infected cells. P170 (MRP) synthesized in this system was capable of carrying out the ATP-dependent transport of leukotriene C4 into isolated membrane vesicles. The results thus indicate that MRP synthesized in insect cells is functional and has properties similar to the authentic protein found overexpressed in certain multidrug-resistant isolates.
Sun
H
H
Division of Biology, Kansas State University, Manhattan 66506, USA.
Zhu
Q
Q
Center
M S
MS
D018528:ATP-Binding Cassette Transporters / Q000096:biosynthesis* / Q000235:genetics; D000255:Adenosine Triphosphate / Q000378:metabolism; D000818:Animals; D002460:Cell Line; D004317:Doxorubicin / Q000494:pharmacology; D018432:Drug Resistance, Multiple / Q000235:genetics*; D019008:Drug Resistance, Neoplasm; D005822:Genetic Vectors / Q000235:genetics*; D006031:Glycosylation; D018922:HL-60 Cells / Q000378:metabolism; D006801:Humans; D017997:Leukotriene C4 / Q000378:metabolism; D027425:ATP-Binding Cassette, Sub-Family C Proteins; D017924:Nucleopolyhedroviruses / Q000235:genetics*; D011499:Protein Processing, Post-Translational; D011993:Recombinant Fusion Proteins / Q000096:biosynthesis* / Q000235:genetics; D018411:Spodoptera / Q000166:cytology
D018528:ATP-Binding Cassette Transporters; D000255:Adenosine Triphosphate; D004317:Doxorubicin; D017997:Leukotriene C4; D027425:ATP-Binding Cassette, Sub-Family C Proteins; D011993:Recombinant Fusion Proteins