Site-directed mutagenesis of Asp-376, the catalytic phosphorylation site, and Lys-507, the putative ATP-binding site, of the alpha-subunit of Torpedo californica Na+/K(+)-ATPase

M Ohtsubo; S Noguchi; K Takeda; M Morohashi; M Kawamura

doi:10.1016/0005-2736(90)90028-m

Site-directed mutagenesis of Asp-376, the catalytic phosphorylation site, and Lys-507, the putative ATP-binding site, of the alpha-subunit of Torpedo californica Na+/K(+)-ATPase

Biochim Biophys Acta. 1990 Jan 29;1021(2):157-60. doi: 10.1016/0005-2736(90)90028-m.

Authors

M Ohtsubo¹, S Noguchi, K Takeda, M Morohashi, M Kawamura

Affiliation

¹ Department of Biology, University of Occupational and Environmental Health, Kitakyushu, Japan.

PMID: 2154258
DOI: 10.1016/0005-2736(90)90028-m

Abstract

Point mutations of Asp-376 of the alpha-subunit of Torpedo californica Na+/K(+)-ATPase (the site of phosphorylation during the catalytic cycle) to Asn, Glu or Thr led to virtual abolishment of Na+/K(+)-ATPase activity and ouabain-binding capacity. Replacement of Lys-507 of the same subunit (the putative ATP-binding site) by Met resulted in decreases in Na+/K(+)-ATPase activity and ouabain-binding capacity. These results are in agreement with those reported for rabbit sarcoplasmic reticulum Ca2(+)-ATPase (Maruyama, K. and MacLennan, D.H. (1988) Proc. Natl. Acad. Sci. USA 85, 3314-3318).

Publication types

Research Support, Non-U.S. Gov't

MeSH terms

Adenosine Triphosphate / metabolism
Animals
Aspartic Acid*
Base Sequence
Binding Sites
Female
Gene Expression
Kinetics
Lysine*
Macromolecular Substances
Molecular Sequence Data
Mutation*
Oligodeoxyribonucleotides / isolation & purification
Oocytes / enzymology
Phosphorylation
Restriction Mapping
Sodium-Potassium-Exchanging ATPase / genetics*
Sodium-Potassium-Exchanging ATPase / metabolism
Torpedo
Xenopus

Substances

Macromolecular Substances
Oligodeoxyribonucleotides
Aspartic Acid
Adenosine Triphosphate
Sodium-Potassium-Exchanging ATPase
Lysine