Functional expression of gastric H,K-ATPase using the baculovirus expression system

FEBS Lett. 1993 Aug 30;329(3):277-82. doi: 10.1016/0014-5793(93)80237-o.

Abstract

A novel approach to construct a single recombinant baculovirus expressing two protein subunits simultaneously by replacing polyhedrin as well as p10 coding sequences is described. The recombinant baculovirus expressed the alpha- as well as the beta-subunit of the gastric H,K-ATPase. Sf9 cells infected with this virus exhibited a K(+)- and SCH 28080-sensitive ATP-dependent phosphorylation capacity in purified Sf9 membranes similar to native H,K-ATPase. This activity was not present in control membranes containing only one of the two H,K-ATPase subunits. We therefore conclude that both subunits are essential for the phosphorylation capacity of H,K-ATPase.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • Baculoviridae / genetics*
  • Base Sequence
  • Cells, Cultured
  • Cloning, Molecular
  • H(+)-K(+)-Exchanging ATPase / genetics
  • H(+)-K(+)-Exchanging ATPase / metabolism*
  • Molecular Sequence Data
  • Moths
  • Oligodeoxyribonucleotides
  • Phosphorylation
  • Rats
  • Stomach / enzymology*

Substances

  • Oligodeoxyribonucleotides
  • H(+)-K(+)-Exchanging ATPase