Determination of P-glycoprotein ATPase activity using luciferase

Tamihide Matsunaga, Eiji Kose, Sachiyo Yasuda, Hirohiko Ise, Uichi Ikeda, Shigeru Ohmori

Research output: Contribution to journalArticle

10 Citations (Scopus)

Abstract

We investigated whether P-glycoprotein (P-gp) ATPase activity of Caco-2 cell membranes could be estimated by measuring consumption of ATP using luciferin-luciferase reaction, and whether the results would be useful for assessment of the interactions between P-gp and drugs. The vanadate-sensitive ATPase activity of Caco-2 cell membranes was measured rapidly with high sensitivity using luciferin-luciferase reaction. Cyclosporin A, verapamil, digoxin and quinidine stimulated the ATPase activity concentration-dependently with Km values of 5.3, 0.9, 1.2 and 4.1 μM, respectively. These values except for digoxin were comparable with previous reports. The ATPase activity and P-gp mRNA expression in Caco-2 cells were induced by all-trans-retinoic acid, digoxin and levothyroxine, but not dexamethasone or rifampicin. This method was useful to assess interactions with P-gp and drugs, and was used to elucidate the mechanisms of interaction of levothyroxine and digoxin.

Original languageEnglish
Pages (from-to)560-564
Number of pages5
JournalBiological and Pharmaceutical Bulletin
Volume29
Issue number3
DOIs
Publication statusPublished - Mar 1 2006

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Digoxin
P-Glycoprotein
Luciferases
Caco-2 Cells
Adenosine Triphosphatases
Thyroxine
Cell Membrane
Rifampin
Tretinoin
Pharmaceutical Preparations
Dexamethasone
Cyclosporine
Adenosine Triphosphate
Messenger RNA

All Science Journal Classification (ASJC) codes

  • Pharmacology
  • Pharmaceutical Science

Cite this

Determination of P-glycoprotein ATPase activity using luciferase. / Matsunaga, Tamihide; Kose, Eiji; Yasuda, Sachiyo; Ise, Hirohiko; Ikeda, Uichi; Ohmori, Shigeru.

In: Biological and Pharmaceutical Bulletin, Vol. 29, No. 3, 01.03.2006, p. 560-564.

Research output: Contribution to journalArticle

Matsunaga, T, Kose, E, Yasuda, S, Ise, H, Ikeda, U & Ohmori, S 2006, 'Determination of P-glycoprotein ATPase activity using luciferase', Biological and Pharmaceutical Bulletin, vol. 29, no. 3, pp. 560-564. https://doi.org/10.1248/bpb.29.560
Matsunaga, Tamihide ; Kose, Eiji ; Yasuda, Sachiyo ; Ise, Hirohiko ; Ikeda, Uichi ; Ohmori, Shigeru. / Determination of P-glycoprotein ATPase activity using luciferase. In: Biological and Pharmaceutical Bulletin. 2006 ; Vol. 29, No. 3. pp. 560-564.
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