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1. Ouabain-sensitive (Na+ + K+)-ATPase activity expressed in mouse L cells by transfection with DNA encoding the alpha-subunit of an avian sodium pump.

Author

Takeyasu K, Tamkun MM, Renaud KJ, and Fambrough DM

Subjects
Amino Acid Sequence, Animals, Base Sequence, Binding Sites, Cell Line, Chick Embryo, Cloning, Molecular, Fibroblasts enzymology, Macromolecular Substances, Mice, Molecular Sequence Data, Rubidium metabolism, Sodium-Potassium-Exchanging ATPase genetics, DNA analysis, Ouabain pharmacology, Sodium-Potassium-Exchanging ATPase metabolism, Transfection
Abstract

cDNA encoding the alpha-subunit of the (Na+ + K+)-ATPase was cloned from a chicken kidney cDNA library and the nucleotide sequence determined. The deduced amino acid sequence showed 92% sequence homology with the alpha-subunit of the sheep kidney (Na+ + K+)-ATPase, and high cross-species homologies were found among nucleotide sequences both in the 5'- and 3'-untranslated regions of the "kidney-type" alpha-subunit mRNAs. The cDNA was subcloned into a shuttle vector derived from pSV2CAT and was stably incorporated into mouse Ltk- cells. Expression of the avian alpha-sub-unit could be activated by culture of the cells in 10 mM butyrate. Cells expressing avian alpha-subunits displayed high-affinity ouabain binding (KD = 2.6 +/- 0.7 x 10(-7) M) and ouabain-sensitive 86Rb+ uptake, characteristic of avian cells.

Published
1988

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