Functional expression of gastric H,K-ATPase using the baculovirus expression system.
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.
Klaassen
C H
CH
Department of Biochemistry, University of Nijmegen, The Netherlands.
Van Uem
T J
TJ
De Moel
M P
MP
De Caluwé
G L
GL
Swarts
H G
HG
De Pont
J J
JJ
D000818:Animals; D016367:Baculoviridae / Q000235:genetics*; D001483:Base Sequence; D002478:Cells, Cultured; D003001:Cloning, Molecular; D017506:H(+)-K(+)-Exchanging ATPase / Q000235:genetics / Q000378:metabolism*; D008969:Molecular Sequence Data; D009036:Moths; D009838:Oligodeoxyribonucleotides; D010766:Phosphorylation; D051381:Rats; D013270:Stomach / Q000201:enzymology*
D009838:Oligodeoxyribonucleotides; D017506:H(+)-K(+)-Exchanging ATPase