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Updated: Oct 23, 2026
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Published on: June 1, 2014
A Simple and Rapid Bioluminescence-Based Functional Assay of Organic Anion Transporter 1 as a D-Luciferin Transporter
Katsuhisa Inoue1, Koki Sugiyama2, Takahito Furuya2
1Department of Biopharmaceutics, School of Pharmacy, Tokyo University of Pharmacy and Life Sciences, Hachioji, Tokyo, Japan. kinoue@toyaku.ac.jp.
Abstract:
Organic anion transporter 1 (SLC22A6/OAT1) plays a key role in renal tubular excretion of endo- and exogenous anionic substances including drugs. Since the inhibition of OAT1 function by a concomitant drug may cause pharmacokinetic drug-drug interactions (DDIs) in clinical practice, an in vitro uptake study to evaluate the inhibition potency of OAT1 is useful for the prediction and avoidance of DDIs and recommended for drug candidates in drug development. In this chapter, we describe a rapid and highly sensitive functional assay of OAT1 based on bioluminescence (BL) detection using D-luciferin as a substrate in living cells. The principle of measurement simply relies on the biochemical feature of D-luciferin to be recognized as a substrate of OAT1, and the BL intensity depending on intracellular D-luciferin level and luciferase activity, thereby allowing the quantitative analysis of OAT1-mediated D-luciferin transport. The BL measurement can be completed within 1 min without experimental procedures for removing extracellular uptake solution and washing cells, both of which involve in the conventional uptake studies using isotope-labeled or fluorescent compounds. The present method is applicable to high-throughput screening to identify and avoid potential OAT1 inhibitors in drug development.
Insights
A novel bioluminescence assay using D-luciferin rapidly measures Organic Anion Transporter 1 (OAT1) activity. This assay aids in predicting drug-drug interactions by identifying OAT1 inhibitors during drug development.
Area of Science:
- Pharmacology
- Biochemistry
- Drug Development
Background:
- Organic Anion Transporter 1 (OAT1) is crucial for renal excretion of anionic substances.
- OAT1 inhibition by co-administered drugs can lead to pharmacokinetic drug-drug interactions (DDIs).
- In vitro OAT1 inhibition assays are vital for predicting and preventing DDIs in drug development.
Purpose of the Study:
- To develop a rapid, sensitive, and high-throughput functional assay for OAT1.
- To utilize bioluminescence detection for quantitative analysis of OAT1-mediated transport.
- To facilitate the identification of OAT1 inhibitors in early drug development.
Main Methods:
- A bioluminescence (BL) assay using D-luciferin as a substrate in living cells.
- Measurement of BL intensity, which correlates with intracellular D-luciferin levels and OAT1 activity.
- OAT1-mediated D-luciferin transport quantified without cell washing or extracellular solution removal.
Main Results:
- The developed BL assay is rapid, completing measurements within 1 minute.
- The assay demonstrates high sensitivity for quantifying OAT1-mediated D-luciferin transport.
- The method is suitable for high-throughput screening of potential OAT1 inhibitors.
Conclusions:
- A novel bioluminescence assay provides a rapid and sensitive method for evaluating OAT1 function.
- This assay is valuable for identifying OAT1 inhibitors and predicting DDIs.
- The assay supports efficient drug development by enabling early screening for OAT1 interactions.
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