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

被引:184
作者
Iliás, A
Urbán, Z
Seidl, TL
Le Saux, O
Sinkó, E
Boyd, CD
Sarkadi, B
Váradi, A
机构
[1] Hungarian Acad Sci, Inst Enzymol, H-1113 Budapest, Hungary
[2] Univ Hawaii, Pacific Biomed Res Ctr, Lab Matrix Pathobiol, Honolulu, HI 96822 USA
[3] Hungarian Acad Sci, Res Grp, Inst Haematol & Immunol, Natl Med Ctr, H-1113 Budapest, Hungary
关键词
D O I
10.1074/jbc.M110918200
中图分类号
Q5 [生物化学]; Q7 [分子生物学];
学科分类号
071010 ; 081704 ;
摘要
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 C-4 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 C-4 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.
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页码:16860 / 16867
页数:8
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