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Articles in PresS, published online ahead of print August 15, 2001
Am J Physiol Renal Physiol, 10.1152/ajprenal.0028.2001
Submitted on February 2, 2001
Accepted on August 2, 2001
1 Medicine, Oregon Health & Science University, Portland, Oregon, USA
* To whom correspondence should be addressed. E-mail: ellisond{at}ohsu.edu.
The thiazide-sensitive Na-Cl cotransporter (NCC) is expressed by distal convoluted tubule cells of the mammalian kidney. We identified by yeast-2-hybrid screening that grp58, a protein induced by glucose deprivation, binds to the carboxyl terminus of the NCC. Immunoprecipitation of rat kidney cortex homogenates using a guinea pig anti-NCC antibody confirmed that grp58 associates with the NCC in vivo. Northern blots indicated that grp58 is highly expressed in rat kidney cortex. Immunofluorescence showed that grp58 protein abundance in kidney is highest in epithelial cells of the distal nephron, where it co-localizes with NCC near the apical membrane. To determine whether this interaction has a functional significance, NCC and grp58 cRNA were co-expressed in Xenopus oocytes. In oocytes over expressing grp58, sodium uptake was increased, compared to control. Because oocytes express endogenous grp58, anti-sense experiments were performed to evaluate whether endogenous grp58 affected NCC activity in oocytes. Sodium uptake was lower in oocytes injected with both anti-sense grp58 cRNA and sense NCC, compared with sense NCC oocytes. Western analysis did not show any effect of grp58 expression on processing of the NCC. These data indicate a novel functionally important interaction between grp58 and the NCC in rat kidney cortex.
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