Thalictrum minus cell cultures and ABC-like transporter.
Cultured Thalictrum minus cells produce a benzylisoquinoline alkaloid, berberine, in the presence of benzyladenine, and excrete it into the culture medium. T. minus cells excluded berberine, even if berberine was exogenously added to the medium, without benzyladenine treatment. Similarly, T. minus cells excluded a heterocyclic dye (neutral red) and calcein AM, which is used as a fluorescent probe to detect the drug efflux pump activity by ABC transporters. The addition of several inhibitors of P-glycoprotein, a representative ABC transporter, induced the accumulation in of both berberine and calcein AM ATP-dependent manner. The expression of P-glycoprotein-like ABC transporter genes was also demonstrated. The involvement of ABC transporter in the secretion of berberine in T. minus cells is discussed.
Terasaka
Kazuyoshi
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.
Sakai
Kyoko
K
Sato
Fumihiko
F
Yamamoto
Hirobumi
H
Yazaki
Kazufumi
K
D020168:ATP Binding Cassette Transporter, Subfamily B, Member 1 / Q000037:antagonists & inhibitors / Q000235:genetics; D018528:ATP-Binding Cassette Transporters / Q000037:antagonists & inhibitors / Q000378:metabolism*; D000595:Amino Acid Sequence; D001599:Berberine / Q000378:metabolism*; D001692:Biological Transport / Q000187:drug effects / Q000502:physiology; D002478:Cells, Cultured; D004587:Electrophoresis, Agar Gel / Q000379:methods; D004791:Enzyme Inhibitors / Q000494:pharmacology; D005452:Fluoresceins; D008856:Microscopy, Fluorescence; D008969:Molecular Sequence Data; D009499:Neutral Red; D018749:RNA, Plant / Q000302:isolation & purification; D020133:Reverse Transcriptase Polymerase Chain Reaction; D017386:Sequence Homology, Amino Acid; D031949:Thalictrum / Q000166:cytology* / Q000187:drug effects / Q000378:metabolism*; D013997:Time Factors
D020168:ATP Binding Cassette Transporter, Subfamily B, Member 1; D018528:ATP-Binding Cassette Transporters; D004791:Enzyme Inhibitors; D005452:Fluoresceins; D018749:RNA, Plant; D001599:Berberine; C085925:calcein AM; D009499:Neutral Red