ATP-Binding Cassette Transporter A Subfamily 8 Is a Sinusoidal Efflux Transporter for Cholesterol and Taurocholate in Mouse and Human Liver

作者：Sasaki Kazunari; Tachikawa Masanori^*; Uchida Yasuo; Hirano Satoshi; Kadowaki Fumito; Watanabe Michitoshi; Ohtsuki Sumio; Terasaki Tetsuya

来源：Molecular Pharmaceutics, 2018, 15(2): 343-355.

DOI：10.1021/acs.molpharmaceut.7b00679

• ABCA6-like subgroup transporters
• ABCA1-like subgroup transporters
• ABCA8
• absolute protein expression level
• tissue distribution
• sinusoidal membrane
• quantitative targeted absolute proteomics
• cholestasis
• liver
• bile duct ligation
• mass Spectrometry

The ATP-binding cassette (ABC) transporter A subfamily 8 (ABCA8) belongs to the ABCA6-like transporters subgroup, which is distinct from the ABCA1-like subgroup in the ABCA family. The expression and function of the short-size human ABCA8 lacking one of the two ATP-binding domains for ATP hydrolysis, which are regularly present in the other ABCA transporters, have been reported. However, the functional differences between the short-size human ABCA8 and full-size human ABCA8, which has the two ATP-binding domains, remain unknown. The, purpose of the present study was to clarify the tissue expression profiles of ABCA6-like and ABCA1-like subgroup transporters and the functional characteristics of ABCA8 in mouse and human. The tissue distribution of mouse ABCA (mABCA) transporter protein and the changes in mABCA8 protein expression levels in a mouse model of obstructive cholestasis were elucidated by means of quantitative targeted absolute proteomics (QTAP). The transport characteristics were clarified in a HEK293 cell line overexpressing full-size ABCA8 protein. QTAP and immunohistochemical analyses revealed that mABCA transporters exhibited the distinct protein expression patterns in the tissues, and mABCA8b, its mouse orthologue was: abundant in the liver and predominantly distributed in sinusoidal membranes of the hepatocytes. Further, protein expression of mABCA8b was decreased in the mouse cholestasis liver. Changes of mABCA8b expression level in cholestasis were similar to those of mABCA1, a sinusoidal cholesterol effhix transporter. Uptake and efflux assays showed that ABCA8 mediates efflux of [H-3]cholesterol and [H-3]taurocholate, while it showed no significant efflux activity for [H-3]estrone sulfate, [H-3]digoxin, [H-3]vinblastine, [H-3]para-aminohippuric acid,[H-3]oleic acid, [C-14]nicotine, or [3H]methotrexate. [H-3] Cholesterolefflux was increased by extracellularly applied taurocholate: These results suggest that mABCA8b/ABCA8 functions as a sinusoidal efflux transporter for at least cholesterol and taurocholate in mouse and human liver.

• 出版日期2018-2