The Journal of Biochemistry
Online ISSN : 1756-2651
Print ISSN : 0021-924X
Characterization, cDNA Cloning, and Functional Expression of Mouse Heal Sodium-Dependent Bile Acid Transporter
Tohru SaekiKeishi MatobaHiroko FurukawaKensuke KirifujiRyuhei KanamotoKimikazu Iwami
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1999 Volume 125 Issue 4 Pages 846-851

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Abstract

Mouse ileal sodium dependent bile acid transporter (ISBT) was characterized using isolated enterocytes. Only enterocytes from the most distal portion showed Na+-dependent [3H]taurocholate uptake. Northern blot analysis using a probe against mouse ISBT revealed the expression of mouse ISBT mRNA to be restricted to the distal ileum. The Km and Vmax, for Na+-dependent [3H]taurocholate transport into isolated ileocytes were calcu-lated as 27 μM and 360 pmol/mg protein/min, respectively. Uptake of [3H]taurocholate was inhibited by N-ethylmaleimide. We have cloned ISBT cDNA from mouse ileum. The cDNA included the entire open reading frame coding 348 amino acid protein with seven hydrophobic segments and two N-glycosylation sites. COS-7 cells transfected with the expression vector containing this cDNA expressed Na+-dependent [3H]taurocholate uptake activity with a Km of

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