Studies on the membrane topology of the (Na,K)-ATPase

Kyunglim L. Yoon, Guido Guidotti

Research output: Contribution to journalArticlepeer-review

37 Scopus citations

Abstract

The topology of the α1 and β1 subunits of the rat (Na,K)-ATPase has been studied by insertion of epitope(s): at the NH2 terminus and COOH terminus and between Glu-117 and Glu-118, Lys-828 and Arg-829, Gln-900 and Trp-901, and Val-939 and Phe-940 of the α subunit; and at the NH2 terminus and COOH terminus and between Glu-228 and Tyr-229 of the β subunit. The epitope- tagged α1 constructs were expressed in HeLa cells to select for stable cell lines expressing a functional (Na,K)-ATPase. The epitope-tagged β constructs were transiently expressed in Cos-7 cells. The membrane arrangement of the epitopes was revealed by indirect immunofluorescence with cells expressing the (Na,K)-ATPase chains. The results indicate that the α subunit has 4 transmembrane segments in the COOH-terminal membrane-bound domain between residues 760 and 938, and that both the NH2 terminus and the COOH terminus are in the cytosol; it was not determined whether there are more transmembrane segments between residue 938 and the COOH terminus. The β subunit has only one transmembrane-spanning region with the NH2 terminus in the cytosol and the COOH terminus on the extracytoplasmic surface of the plasma membrane.

Original languageEnglish
Pages (from-to)28249-28258
Number of pages10
JournalJournal of Biological Chemistry
Volume269
Issue number45
StatePublished - 11 Nov 1994

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