26 Amino acids of an extracellular domain of the Na,K-ATPase α-subunit are sufficient for assembly with the Na,K-ATPase β-subunit

M. Victor Lemas, Maura Hamrick, Kunio Takeyasu, Douglas M. Fambrough

Research output: Contribution to journalArticlepeer-review

Abstract

Chimeric cDNAs encoding a sarcoplasmic/endoplasmic reticulum Ca-ATPase (SERCA1) and regions of the Na,K-ATPase α-subunit were constructed to seek the minimal region of the α-subunit sufficient for assembly with the Na,K- ATPase β-subunit. cDNAs encoding a chimera and the chicken β-subunit were coexpressed in mammalian cells and assembly was assayed by immune precipitation of the chimeric subunit with a monoclonal antibody to the chicken β-subunit. A chimera containing 26 amino acyl residues of the Na,K- ATPase α1-subunit (NDVEDSYGQQWTFEQRKIVEFTCHTA) (Asn894 to Ala919) that replaced the corresponding avian SERCA1 Ca-ATPase amino acyl residues (Thr871 to Thr898) was able to assemble with the chicken β-subunit. This α-subunit region is predicted to be extracellular, located between membrane-spanning domains 7 and 8 (H7-H8). Chimeras that assembled with full- length β-subunit also assembled with a β-subunit chimera that retained only the ectodomain of the chicken β1-subunit. These results suggest that the Na,K-ATPase α-subunit has the same topology in the membrane as the sarcoplasmic reticulum Ca-ATPase, probably with 10 membrane-spanning domains, and that the aminoacyl residues between membrane domains H7 and H8 are involved in assembly with the β-subunit in the extracellular/lumenal space.

Original languageEnglish (US)
Pages (from-to)8255-8259
Number of pages5
JournalJournal of Biological Chemistry
Volume269
Issue number11
StatePublished - Mar 18 1994

ASJC Scopus subject areas

  • Biochemistry
  • Molecular Biology
  • Cell Biology

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