title: Active plasma membrane P-type H+-ATPase reconstituted into nanodiscs is a monomer.
abstract: Plasma membrane H(+)-ATPases form a subfamily of P-type ATPases responsible for pumping protons out of cells and are essential for establishing and maintaining the crucial transmembrane proton gradient in plants and fungi. Here, we report the reconstitution of the Arabidopsis thaliana plasma membrane H(+)-ATPase isoform 2 into soluble nanoscale lipid bilayers, also termed nanodiscs. Based on native gel analysis and cross-linking studies, the pump inserts into nanodiscs as a functional monomer. Insertion of the H(+)-ATPase into nanodiscs has the potential to enable structural and functional characterization using techniques normally applicable only for soluble proteins.
authors.affiliation: From the Centre for Membrane Pumps in Cells and Disease-PUMPKIN.
authors.forename: Bo Højen
authors.initials: BH
authors.lastname: Justesen
authors.forename: Randi Westh
authors.initials: RW
authors.lastname: Hansen
authors.forename: Helle Juel
authors.initials: HJ
authors.lastname: Martens
authors.forename: Lisa
authors.initials: L
authors.lastname: Theorin
authors.forename: Michael G
authors.initials: MG
authors.lastname: Palmgren
authors.forename: Karen L
authors.initials: KL
authors.lastname: Martinez
authors.forename: Thomas Günther
authors.initials: TG
authors.lastname: Pomorski
authors.forename: Anja Thoe
authors.initials: AT
authors.lastname: Fuglsang
mesh_terms: D017360:Arabidopsis / Q000201:enzymology*; D029681:Arabidopsis Proteins / Q000378:metabolism*; D002462:Cell Membrane / Q000201:enzymology*; D003432:Cross-Linking Reagents; D004789:Enzyme Activation; D004926:Escherichia coli / Q000378:metabolism; D007527:Isoenzymes / Q000378:metabolism; D008051:Lipid Bilayers / Q000378:metabolism*; D046529:Microscopy, Electron, Transmission; D006180:Proton-Translocating ATPases / Q000378:metabolism*; D012441:Saccharomyces cerevisiae / Q000378:metabolism; D020349:Surface Plasmon Resonance
chemical_list: D029681:Arabidopsis Proteins; D003432:Cross-Linking Reagents; D007527:Isoenzymes; D008051:Lipid Bilayers; D006180:Proton-Translocating ATPases; C479903:AHA2 protein, Arabidopsis
keywords: H+-ATPase; Monomer; Nanodiscs; Plasma Membrane Proton Pumps; Reconstitution of Membrane Transporters; Scaffold Proteins; Surface Plasmon Resonance (SPR)