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Journal of Biomolecular Screening
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Mouse Bsep ATPase Assay: A Nonradioactive Tool for Assessment of the Cholestatic Potential of Drugs

Emese Kis

Solvo Biotechnology, Szeged, Hungary

Zsuzsanna Rajnai

Solvo Biotechnology, Szeged, Hungary

Eniko Ioja

Solvo Biotechnology, Szeged, Hungary

Krisztina Herédi Szabó

Solvo Biotechnology, Szeged, Hungary

Tünde Nagy

Solvo Biotechnology, Szeged, Hungary

Dóra Méhn

Solvo Biotechnology, Szeged, Hungary

Péter Krajcsi

Solvo Biotechnology, Szeged, Hungary, krajcsi{at}solvo.com

The mouse ortholog of the human bile salt export pump (BSEP) transporter was expressed in a baculovirus-infected insect cell (Sf9) system to study the effect of membrane cholesterol content on the transporter function. The transport activity of cholesterol-loaded mouse Bsep-HAM-Sf9 vesicles was determined in a vesicular transport assay with taurochenodeoxycholate (TCDC), a known BSEP substrate. Mouse Bsep transports TCDC at a high rate that can be sensitively detected in the ATPase assay. Cholesterol upload of the Sf9 membrane potentiates both TCDC transport and TCDC-stimulated ATPase activities. Inhibitory effect of BSEP interactors on probe substrate transport was tested in both vesicular transport and ATPase assays using cholesterol-loaded membrane vesicles. A good rank order correlation was found between IC50 values measured in TCDC-stimulated mBsep ATPase assay and in the human BSEP vesicular transport assay utilizing taurocholate (TC) as probe substrate. This upgraded form of the mouse Bsep-HAM ATPase assay is a user friendly, sensitive, nonradioactive method for early high-throughput screening of drugs with BSEP-related cholestatic potential. It may complement the human BSEP-mediated taurocholate vesicular transport inhibition assay. (Journal of Biomolecular Screening 2009:10-15)

Key Words: mouse • Bsep/ABCB11 • cholestasis • ATPase activity • nonradioactive • taurochenodeoxycholate

This version was published on January 1, 2009

Journal of Biomolecular Screening, Vol. 14, No. 1, 10-15 (2009)
DOI: 10.1177/1087057108326145
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 Drug Metab. Dispos.Home page
E. Kis, E. Ioja, T. Nagy, L. Szente, K. Heredi-Szabo, and P. Krajcsi
Effect of Membrane Cholesterol on BSEP/Bsep Activity: Species Specificity Studies for Substrates and Inhibitors
Drug Metab. Dispos., September 1, 2009; 37(9): 1878 - 1886.
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