Comparison of 3 assay systems using a common probe substrate, calcein AM, for studying P-gp using a selected set of compounds

Péter Szerémy, Ákos Pál, Dóra Méhn, Beáta Tóth, F. Fülöp, Péter Krajcsi, Krisztina Herédi-Szabó

Research output: Contribution to journalArticle

15 Citations (Scopus)

Abstract

The multidrug resistance protein 1 (MDR1) transporter is the most abundantly investigated adenosine triphosphate (ATP)- Binding Cassette (ABC) transporter protein. Multiple assay systems were developed to study MDR1-mediated transport and possible drug-drug interactions. Yet, as different probe substrates are used in these assays, it is difficult to directly compare the results. In this study, a common probe substrate was applied in 3 assay systems developed to study MDR1: the cellular dye efflux assay, the ATPase assay, and the vesicular transport assay. This probe substrate is calcein acetoxymethyl ester (calcein AM), the acetoxymethyl ester derivative of the fluorescent dye, calcein. Using a common probe allows the investigation of the effect of passive permeability on the result obtained by testing various compounds. In this study, 22 compounds with different logP values were tested in the above-mentioned 3 assay systems. The vesicular transport assay proved most sensitive, detecting 18 of 22 interactions with the protein. The ATPase assay detected 15 interactions, whereas the cellular dye efflux assay was the least sensitive with only 10 hits. A correlation was found between the hydrophobicity of the compound and the ratio of cellular and vesicular transport IC 50 values, indicating the effect of passive permeability on the result. Based on hydrophobicity, the current study provides guidelines on applying the most correct tool for studying MDR1 interactions.

Original languageEnglish
Pages (from-to)112-119
Number of pages8
JournalJournal of Biomolecular Screening
Volume16
Issue number1
DOIs
Publication statusPublished - Jan 2011

Fingerprint

P-Glycoprotein
Assays
Substrates
Hydrophobic and Hydrophilic Interactions
Adenosine Triphosphatases
Permeability
Coloring Agents
Drug Interactions
Fluorescent Dyes
Hydrophobicity
Esters
Proteins
Adenosine Triphosphate
Guidelines
calcein AM
Drug interactions
Pharmaceutical Preparations
Derivatives
Testing

Keywords

  • calcein AM
  • correlation
  • in vitro
  • logP
  • MDR1

ASJC Scopus subject areas

  • Analytical Chemistry
  • Drug Discovery
  • Pharmacology
  • Biochemistry
  • Molecular Medicine
  • Biotechnology

Cite this

Comparison of 3 assay systems using a common probe substrate, calcein AM, for studying P-gp using a selected set of compounds. / Szerémy, Péter; Pál, Ákos; Méhn, Dóra; Tóth, Beáta; Fülöp, F.; Krajcsi, Péter; Herédi-Szabó, Krisztina.

In: Journal of Biomolecular Screening, Vol. 16, No. 1, 01.2011, p. 112-119.

Research output: Contribution to journalArticle

Szerémy, Péter ; Pál, Ákos ; Méhn, Dóra ; Tóth, Beáta ; Fülöp, F. ; Krajcsi, Péter ; Herédi-Szabó, Krisztina. / Comparison of 3 assay systems using a common probe substrate, calcein AM, for studying P-gp using a selected set of compounds. In: Journal of Biomolecular Screening. 2011 ; Vol. 16, No. 1. pp. 112-119.
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