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Radanovic, T; Murer, H; Biber, J (2003). Expression of the Na/P(i)-cotransporter type IIb in Sf9 cells: functional characterization and purification. Journal of Membrane Biology, 194(2):91-96.

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Abstract

In mammals the type IIb Na/P(i)-cotransporter is expressed in various tissues such as intestine, brain, lung and testis. The type IIb cotransporter shows 51% homology with the renal type IIa Na/P(i)-cotransporter, for which a detailed model of the secondary structure has emerged based on recent structure/function studies. To make the type IIb Na/P(i)-cotransporter available for future structural studies, we have expressed this cotransporter in Sf9 cells. Sf9 cells were infected with recombinant baculovirus containing 6His NaPi-IIb. Infected cells expressed a polypeptide of approximately 90 kDa, corresponding to a partially glycosylated form of the type IIb cotransporter. Transport studies demonstrated that the type IIb protein expressed in Sf9 cells mediates transport of phosphate in a Na-dependent manner with similar kinetic characteristics (apparent K(m)s for sodium and phosphate and pH dependence) as previously described. Solubilization experiments demonstrated that, in contrast to the type IIa cotransporter, the type IIb can be solubilized by nonionic detergents and that solubilized type IIb Na/P(i)-cotransporter can be purified by Ni-NTA chromatography.

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5 citations in Web of Science®
6 citations in Scopus®
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Additional indexing

Item Type:Journal Article, refereed
Communities & Collections:04 Faculty of Medicine > Institute of Physiology
07 Faculty of Science > Institute of Physiology
DDC:570 Life sciences; biology
Language:English
Date:15 July 2003
Deposited On:11 Feb 2008 12:22
Last Modified:27 Nov 2013 18:23
Publisher:Springer
ISSN:0022-2631
Publisher DOI:10.1007/s00232-003-2028-6
PubMed ID:14502433

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