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Journal of Experimental Botany, Vol. 53, No. 376, pp. 1879-1886, September 1, 2002
© 2002 Oxford University Press

Characterization of berberine transport into Coptis japonica cells and the involvement of ABC protein

Received 4 March 2002; Accepted 11 June 2002

Kyoko Sakai1, Nobukazu Shitan1, Fumihiko Sato1, Kazumitsu Ueda2 and Kazufumi Yazaki3,1

1 Laboratory of Molecular and Cellular Biology of Totipotency, Division of Integrated Life Science, Graduate School of Biostudies, Kyoto University, Kitashirakawa, Kyoto 606-8502, Japan
2 Laboratory of Cellular Biochemistry, Division of Applied Life Sciences, Graduate School of Agriculture, Kyoto University, Kitashirakawa, Kyoto 606-8502, Japan

Abbreviations: ABC, ATP-binding cassette; DMSO, dimethylsulphoxide; DTT, dithiothreitol; EGTA, ethylene glycol bis(2-aminoethyl)-tetra-acetic acid; MDR, multi-drug resistance protein; MRP, multi-drug resistance-associated 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.

Key words: Key words: ABC protein, berberine, Coptis japonica, MDR, transport.


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