Klaassen C H, Swarts H G, De Pont J J
Department of Biochemistry, University of Nijmegen, The Netherlands.
Biochem Biophys Res Commun. 1995 May 25;210(3):907-13. doi: 10.1006/bbrc.1995.1743.
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.
杆状病毒表达系统适用于胃H⁺，K⁺-ATP酶的功能表达。在Sf9细胞中功能性H⁺，K⁺-ATP酶的表达伴随着大量无功能亚基的合成。当在培养基中150 - 250 mM乙醇存在的情况下合成H⁺，K⁺-ATP酶时，由于形成了更多的功能亚基而非亚基本身数量的增加，功能性H⁺，K⁺-ATP酶的产量提高了两到三倍。乙醇产生的H⁺，K⁺-ATP酶的催化特性与对照制剂没有区别。乙醇刺激功能性H⁺，K⁺-ATP酶形成的机制可能涉及对Sf9膜物理性质的直接影响。此外，乙醇诱导的应激蛋白作为分子伴侣也可能产生间接影响。
