title: Expression of Na,K-ATPase and H,K-ATPase Isoforms with the Baculovirus Expression System.
abstract: P-type ATPases can be expressed in several cell systems. The baculovirus expressions system uses an insect virus to enter and express proteins in Sf9 insect cells. This expression system is a lytic system in which the cells will die a few days after viral infection. Subsequently, the expressed proteins can be isolated. Insect cells are a perfect system to study P-type ATPases as they have little or no endogenous Na,K-ATPase activity and other ATPase activities can be inhibited easily. Here we describe in detail the expression and isolation of Na,K-ATPase and H,K-ATPase isoforms with the baculovirus expression system.
authors.affiliation: Pharmacology/Toxicology 149, Radboud University Medical Center, Radboud Institute for Molecular Life Sciences, 9101, 6500 HB, Nijmegen, The Netherlands. Jan.Koenderink@radboudumc.nl.
authors.forename: Jan B
authors.initials: JB
authors.lastname: Koenderink
authors.affiliation: Radboud University Medical Center, Nijmegen, The Netherlands.
authors.forename: Herman G P
authors.initials: HG
authors.lastname: Swarts
mesh_terms: D000818:Animals; D016367:Baculoviridae / Q000235:genetics; D002462:Cell Membrane / Q000737:chemistry / Q000201:enzymology; D015967:Gene Expression Regulation, Viral; D017506:H(+)-K(+)-Exchanging ATPase / Q000096:biosynthesis / Q000235:genetics / Q000302:isolation & purification*; D007313:Insecta / Q000201:enzymology; D008967:Molecular Biology / Q000379:methods*; D020033:Protein Isoforms / Q000096:biosynthesis / Q000235:genetics / Q000302:isolation & purification*; D000254:Sodium-Potassium-Exchanging ATPase / Q000096:biosynthesis / Q000235:genetics / Q000302:isolation & purification*
chemical_list: D020033:Protein Isoforms; D017506:H(+)-K(+)-Exchanging ATPase; D000254:Sodium-Potassium-Exchanging ATPase
keywords: Baculovirus; H,K-ATPase; Insect cells; Isoforms; Membrane isolation; Na,K-ATPase; Sf9 cells