title: Measurement of transport activities of 3β-hydroxy-Δ(5)-bile acids in bile salt export pump and multidrug resistance-associated proteins using LC-MS/MS.
abstract: A method has been developed for the measurement of transport activities in membrane vesicles obtained from Sf9 cells for 3β-hydroxy-Δ(5)-bile acids by high-performance liquid chromatography-electrospray ionization-tandem mass spectrometry. Calibration curves for the bile acids were linear over the range of 10 to 2000 pmol/mL, and the detection limit was less than 1 pmol/mL for 3β-hydroxy-Δ(5)-bile acids using selected reaction monitoring analysis. The analytical method was applied to measurements of transport activities in membrane vesicles obtained from human multidrug resistance-associated protein 2-, 3-, and human bile salt export pump-expressing Sf9 cells for conjugated 3β-hydroxy-Δ(5)-bile acids. The present study demonstrated that human multidrug resistance-associated protein 3 vesicles accepted conjugated 3β-hydroxy-Δ(5)-bile acids along with common bile acids such as glycocholic acid and taurolithocholic acid 3-sulfate.
authors.affiliation: School of Pharmaceutical Sciences, Health Sciences University of Hokkaido, Kanazawa, Ishikari-Tobetsu, Hokkaido 061-0293, Japan. murai@hoku-iryo-u.ac.jp
authors.forename: Tsuyoshi
authors.initials: T
authors.lastname: Murai
authors.forename: Kana
authors.initials: K
authors.lastname: Oda
authors.forename: Terutake
authors.initials: T
authors.lastname: Toyo
authors.forename: Hiroshi
authors.initials: H
authors.lastname: Nittono
authors.forename: Hajime
authors.initials: H
authors.lastname: Takei
authors.forename: Akina
authors.initials: A
authors.lastname: Muto
authors.forename: Akihiko
authors.initials: A
authors.lastname: Kimura
authors.forename: Takao
authors.initials: T
authors.lastname: Kurosawa
mesh_terms: D000818:Animals; D001647:Bile Acids and Salts / Q000032:analysis* / Q000737:chemistry* / Q000378:metabolism; D002851:Chromatography, High Pressure Liquid; D006801:Humans; D008968:Molecular Conformation; D027425:ATP-Binding Cassette, Sub-Family C Proteins / Q000737:chemistry* / Q000378:metabolism; D061987:Sf9 Cells; D018411:Spodoptera; D053719:Tandem Mass Spectrometry
chemical_list: D001647:Bile Acids and Salts; D027425:ATP-Binding Cassette, Sub-Family C Proteins