Topology of Na,K-ATPase α subunit epitopes analyzed with oligopeptide-specific antibodies and double-labeling immunoelectron microscopy

Gang Ning, Arvid B. Maunsbach, Yang Jae Lee, Jesper Vuust Møller

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21 Scopus citations

Abstract

Using four oligopeptide-specific polyclonal antibodies, we mapped the α subunit ofNa,K-ATPase by double-labeling immunoelectron microscopy combined with negative staining. The results show that the epitopes of the N-tenninus (Gly1-His13), C-terminus (Ile1002-Tyr1016) and Leu815-Gln828 are located on the same face of crystallized Na,K-ATPase membranes from pig kidney, whereas the epitope Asn889-Gln903 is present on the opposite side. The present study demonstrates the cytoplasmic location of C-terminus and that Leu828-Gln828 is exposed on the eytoplasmic and Asn889-Gln903 on the extracellular side. The results are consistent with an eight- or ten-segment model, and support the existence of an M5/M6 loop and the presence of one transmembrane segment between Leu815-Gln828 and Asn889-Gln903.

Original languageEnglish (US)
Pages (from-to)521-524
Number of pages4
JournalFEBS Letters
Volume336
Issue number3
DOIs
StatePublished - Dec 28 1993

All Science Journal Classification (ASJC) codes

  • Biophysics
  • Structural Biology
  • Biochemistry
  • Molecular Biology
  • Genetics
  • Cell Biology

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