The human multidrug-resistance-associated protein MRP1 mediates ATP-dependent transport of unconjugated bilirubin

Igino Rigato, Lorella Pascolo, Cristina Fernetti, J. Donald Ostrow, Claudio Tiribelli

Research output: Contribution to journalArticle

50 Citations (Scopus)

Abstract

Results of previous studies have suggested that UCB (unconjugated bilirubin) may be transported by MRP1/Mrp1 (multidrug-resistance-associated protein 1). To test this hypothesis directly, [3H]UCB transport was assessed in plasma-membrane vesicles from MDCKII cells (Madin-Darby canine kidney II cells) stably transfected with human MRP1 or MRP2; wild-type MDCKII cells served as controls. As revealed by Western blotting, transfection achieved abundant expression of MRP1 and MRP2. [3H]UCB uptake was measured in the presence of 60 μM human serum albumin at a free (unbound) concentration of UCB (BF) ranging from 5 to 72 nM and in the presence of 3 mM ATP or 3 mM AMP-PCP (adenosine 5′-[β,γ-methylene]triphosphate). MRP1-transfected vesicles showed transport activity three and five times higher respectively compared with MRP2 or wildtype vesicles, whose transport did not differ significantly. [3H]UCB transport was stimulated 4-fold by 1.5 mM GSH, occurred into an osmotically sensitive space, was inhibited by 3 μM MK571 and followed saturative kinetics with Km = 10 ± 3 nM (BF) and Vmax = 100 ± 13 pmol · min -1 · (mg of protein)-1. UCB significantly inhibited the transport of LTC4 (leukotriene C4), a leukotriene substrate known to have high affinity for MRP1. Collectively, these results prove directly that MRP1 mediates ATP-dependent cellular export of UCB and supports its role in protecting cells from bilirubin toxicity.

Original languageEnglish
Pages (from-to)335-341
Number of pages7
JournalBiochemical Journal
Volume383
Issue number2
DOIs
Publication statusPublished - Oct 15 2004

Fingerprint

Multidrug Resistance-Associated Proteins
Bilirubin
Adenosine Triphosphate
Transport Vesicles
Leukotriene C4
Madin Darby Canine Kidney Cells
Leukotrienes
Cell membranes
Serum Albumin
Adenosine
Transfection
Toxicity
Western Blotting
Cell Membrane
Kinetics

Keywords

  • ATP-binding-cassette protein (ABC) protein
  • Cytotoxicity
  • Glutathione
  • Multidrug-resistance-associated protein 1 (MRP1)
  • Transport
  • Unconjugated bilirubin

ASJC Scopus subject areas

  • Biochemistry

Cite this

The human multidrug-resistance-associated protein MRP1 mediates ATP-dependent transport of unconjugated bilirubin. / Rigato, Igino; Pascolo, Lorella; Fernetti, Cristina; Ostrow, J. Donald; Tiribelli, Claudio.

In: Biochemical Journal, Vol. 383, No. 2, 15.10.2004, p. 335-341.

Research output: Contribution to journalArticle

Rigato, Igino ; Pascolo, Lorella ; Fernetti, Cristina ; Ostrow, J. Donald ; Tiribelli, Claudio. / The human multidrug-resistance-associated protein MRP1 mediates ATP-dependent transport of unconjugated bilirubin. In: Biochemical Journal. 2004 ; Vol. 383, No. 2. pp. 335-341.
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