Characterization of berberine transport into Coptis japonica cells and the involvement of ABC protein.
Cultured Coptis japonica cells are able to take up berberine, a benzylisoquinoline alkaloid, from the medium and transport it exclusively into the vacuoles. Uptake activity depends on the growth phase of the cultured cells whereas the culture medium had no effect on uptake. Treatment with several inhibitors suggested that berberine uptake depended on the ATP level. Some inhibitors of P-glycoprotein, an ABC transporter involved in multiple drug resistance in cancer cells, strongly inhibited berberine uptake, whereas a specific inhibitor for glutathione biosynthesis and vacuolar ATPase, bafilomycin A1, had little effect. Vanadate-induced ATP trap experiments to detect ABC proteins expressed in C. japonica cells showed that three membrane proteins of between 120 and 150 kDa were photolabelled with 8-azido-[alpha-32P] ATP. Two revealed the same photoaffinity-labelling pattern as P-glycoprotein, and the interaction of these proteins with berberine was also demonstrated. These results suggest that ABC proteins of the MDR-type are involved in the uptake of berberine from the medium.
Sakai
Kyoko
K
Laboratory of Molecular and Cellular Biology of Totipotency, Division of Integrated Life Science, Graduate School of Biostudies, Kyoto University, Kitashirakawa, Kyoto 606-8502, Japan.
Shitan
Nobukazu
N
Sato
Fumihiko
F
Ueda
Kazumitsu
K
Yazaki
Kazufumi
K
D018528:ATP-Binding Cassette Transporters / Q000037:antagonists & inhibitors / Q000302:isolation & purification / Q000378:metabolism*; D000900:Anti-Bacterial Agents / Q000494:pharmacology; D001386:Azides / Q000494:pharmacology; D001599:Berberine / Q000378:metabolism*; D001692:Biological Transport / Q000187:drug effects / Q000502:physiology; D002478:Cells, Cultured; D016572:Cyclosporine / Q000494:pharmacology; D019450:Genes, MDR / Q000187:drug effects; D006863:Hydrogen-Ion Concentration; D029684:Lithospermum / Q000166:cytology / Q000187:drug effects / Q000378:metabolism; D018942:Macrolides; D009543:Nifedipine / Q000494:pharmacology; D011802:Quinidine / Q000494:pharmacology; D029626:Ranunculaceae / Q000166:cytology / Q000187:drug effects / Q000378:metabolism*; D014026:Nicotiana / Q000166:cytology / Q000187:drug effects / Q000378:metabolism; D014617:Vacuoles / Q000502:physiology; D014638:Vanadates / Q000494:pharmacology
D018528:ATP-Binding Cassette Transporters; D000900:Anti-Bacterial Agents; D001386:Azides; D018942:Macrolides; D001599:Berberine; D014638:Vanadates; D016572:Cyclosporine; C040929:bafilomycin A1; D009543:Nifedipine; D011802:Quinidine