Ethanol stimulates expression of functional H+,K(+)-ATPase in SF9 cells.
The baculovirus expression system is suitable for functional expression of gastric H+,K(+)-ATPase. Expression of functional H+,K(+)-ATPase in Sf9 cells is accompanied by synthesis of large amounts of non-functional subunits. When H+,K(+)-ATPase is synthesised in the presence of 150-250 mM ethanol in the culture medium, two to threefold higher levels of functional H+,K(+)-ATPase are produced due to the formation of more functional subunits rather than to an increase of subunits per se. The catalytical properties of the ethanol-produced H+,K(+)-ATPase are indistinguishable from control preparations. The mechanism by which ethanol stimulates the formation of functional H+,K(+)-ATPase probably involves a direct effect on the physical properties of Sf9 membranes. In addition there also might be an indirect effect through ethanol inducible stress proteins acting as molecular chaperones.
Klaassen
C H
CH
Department of Biochemistry, University of Nijmegen, The Netherlands.
Swarts
H G
HG
De Pont
J J
JJ
D000438:Alcohols / Q000494:pharmacology; D000818:Animals; D002460:Cell Line; D002462:Cell Membrane / Q000201:enzymology; D000431:Ethanol / Q000494:pharmacology*; D005753:Gastric Mucosa / Q000201:enzymology; D015870:Gene Expression / Q000187:drug effects*; D017506:H(+)-K(+)-Exchanging ATPase / Q000096:biosynthesis* / Q000378:metabolism; D007700:Kinetics; D011994:Recombinant Proteins / Q000096:biosynthesis / Q000378:metabolism; D018411:Spodoptera; D013329:Structure-Activity Relationship; D013552:Swine; D014162:Transfection
D000438:Alcohols; D011994:Recombinant Proteins; D000431:Ethanol; D017506:H(+)-K(+)-Exchanging ATPase