Loss of ATP-dependent transport activity in pseudoxanthoma elasticum-associated mutants of human ABCC6 (MRP6)

Attila Iliás, Zsolt Urbán, Thomas L. Seidl, Olivier Le Saux, Emese Sinkó, Charles D. Boyd, B. Sarkadi, A. Váradi

Research output: Contribution to journalArticle

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Abstract

Mutations in the ABCC6 (MRP6) gene cause pseudoxanthoma elasticum (PXE), a rare heritable disorder resulting in the calcification of elastic fibers. In the present study a cDNA encoding a full-length normal variant of ABCC6 was amplified from a human kidney cDNA library, and the protein was expressed in Sf9 insect cells. In isolated membranes ATP binding as well as ATP-dependent active transport by ABCC6 was demonstrated. We found that glutathione conjugates, including leukotriene C4 and N-ethylmaleimide S-glutathione (NEM-GS), were actively transported by human ABCC6. Organic anions (probenecid, benzbromarone, indomethacin), known to interfere with glutathione conjugate transport of human ABCC1 and ABCC2, inhibited the ABCC6-mediated NEM-GS transport in a specific manner, indicating that ABCC6 has a unique substrate specificity. We have also expressed three missense mutant forms of ABCC6, which have recently been shown to cause PXE. MgATP binding was normal in these proteins; ATP-dependent NEM-GS or leukotriene C4 transport, however, was abolished. Our data indicate that human ABCC6 is a primary active transporter for organic anions. In the three ABCC6 mutant forms examined, the loss of transport activity suggests that these mutations result in a PXE phenotype through a direct influence on the transport activity of this ABC transporter.

Original languageEnglish
Pages (from-to)16860-16867
Number of pages8
JournalJournal of Biological Chemistry
Volume277
Issue number19
DOIs
Publication statusPublished - May 10 2002

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Pseudoxanthoma Elasticum
Rubber
Adenosine Triphosphate
Leukotriene C4
Glutathione
Benzbromarone
Organic Anion Transporters
Sf9 Cells
Probenecid
Mutation
Elastic Tissue
ATP-Binding Cassette Transporters
Active Biological Transport
Substrate Specificity
Gene Library
Indomethacin
Anions
Insects
Proteins
Complementary DNA

ASJC Scopus subject areas

  • Biochemistry

Cite this

Loss of ATP-dependent transport activity in pseudoxanthoma elasticum-associated mutants of human ABCC6 (MRP6). / Iliás, Attila; Urbán, Zsolt; Seidl, Thomas L.; Saux, Olivier Le; Sinkó, Emese; Boyd, Charles D.; Sarkadi, B.; Váradi, A.

In: Journal of Biological Chemistry, Vol. 277, No. 19, 10.05.2002, p. 16860-16867.

Research output: Contribution to journalArticle

Iliás, Attila ; Urbán, Zsolt ; Seidl, Thomas L. ; Saux, Olivier Le ; Sinkó, Emese ; Boyd, Charles D. ; Sarkadi, B. ; Váradi, A. / Loss of ATP-dependent transport activity in pseudoxanthoma elasticum-associated mutants of human ABCC6 (MRP6). In: Journal of Biological Chemistry. 2002 ; Vol. 277, No. 19. pp. 16860-16867.
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