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Expression cloning of rat renal Na+/SO4(2-) cotransport.
Markovich D
,
Forgo J
,
Stange G
,
Biber J
,
Murer H
.
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Injection of rat kidney cortex mRNA into Xenopus laevis oocytes leads to a stimulation of Na(+)-dependent SO4(2-) uptake. Based on this information, we have isolated from a corresponding library a cDNA (NaSi-1) that is most likely related to a Na+/SO4(2-) cotransport system. NaSi-1 cRNA leads in a time- and dose-dependent manner to specific stimulation of Na(+)-dependent SO4(2-) uptake in oocytes. The apparent affinity constants of the NaSi-1 cRNA-expressed transport resemble those of Na+/SO4(2-) cotransport in brush-border membrane. The NaSi-1 cDNA contains 2239 bp [including a poly(A) tail] and encodes a protein of 595 amino acids (66.05 kDa); the hydropathy profile suggests at least eight membrane-spanning regions. In vitro translation of NaSi-1 cRNA results in a protein of the expected size and suggests glycosylation. Northern blot analysis shows signals of 2.3 and 2.9 kb in kidney (more abundant in cortex than in papilla/medulla) and in mucosa of small intestine of rats. The above data indicate that we have structurally identified a membrane protein involved in renal and small-intestinal brush-border membrane Na+/SO4(2-) cotransport.
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