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J. Biochem, 2003, Vol. 134, No. 2 225-230
© 2003 Japanese Biochemical Society

BIOCHEMISTRY

ATP-Dependent Transport of Bile Acid Intermediates across Rat Liver Peroxisomal Membranes

Mizuho Une3,*, Yusuke Iguchi1, Tomoko Sakamoto1, Takashi Tomita1, Yasuyuki Suzuki2, Masashi Morita3 and Tsuneo Imanaka

1 Division of Medical Chemistry, Programs for Pharmaceutical Sciences, Graduate School of Biomedical Sciences, Hiroshima University, Kasumi, Minami-ku, Hiroshima 734-8551; 2 Department of Pediatrics, Gifu University School of Medicine, Tukasa, Gifu 500-8076; and 3 Department of Biological Chemistry, Faculty of Pharmaceutical Sciences, Toyama Medical and Pharmaceutical University, 2630 Sugitani, Toyama 930-0194

The bile acid intermediate 3{alpha},7{alpha},12{alpha}-trihydroxy-5ß-cholestanoic acid (THCA) is converted to cholic acid exclusively in peroxisomes by the oxidative cleavage of the side chain. To investigate the mechanism by which the biosynthetic intermediates of bile acids are transported into peroxisomes, we incubated THCA or its CoA ester (THC-CoA) with isolated intact rat liver peroxisomes and analyzed their oxidation products, cholic acid and 3{alpha},7{alpha},12{alpha}-trihydroxy-5ß-cholest-24-enoic acid. The oxidation of both THCA and THC-CoA was dependent on incubation time and peroxisomal proteins, and was stimulated by ATP. THC-CoA was efficiently oxidized to cholic acid and 3{alpha},7{alpha},12{alpha}-trihydroxy-5ß-cholest-24-enoic acid as compared with THCA, suggesting that THC-CoA is the preferred substrate for transport into peroxisomes. The oxidation of THC-CoA was significantly inhibited by sodium azide, verapamile, and N-ethylmaleimide. Furthermore, the stimulatory effect of ATP on the oxidation was not replaced by GTP or AMP. In addition, the ATP-dependent oxidation of THC-CoA was markedly inhibited by pretreatment of peroxisomes with proteinase K when peroxisomal matrix proteins were not degraded. These results suggest that an ATP-dependent transport system for THC-CoA exists on peroxisomal membranes.

* To whom correspondence should be addressed. Tel: +81-82-257-5297, E-mail: mune{at}hiroshima-u.ac.jp


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