title: Proton transport through a peptide-tethered bilayer lipid membrane by the H(+)-ATP synthase from chloroplasts measured by impedance spectroscopy.
abstract: A lipid membrane was tethered to a gold film by a peptide spacer molecule terminated by a sulfhydryl group. Membranes were formed by fusion of liposomes prepared from egg phosphatidylcholine on self assembled monolayers of the thiolipopeptide Myr-Lys(Myr)-Ser-Ser-Pro-Ala-Ser-Ser-Ala-Ala-Ser-Ala-Cys-amide mixed with mercaptoethanol as a diluent molecule or lateral spacer. These mixed films, although not representing a perfect lipid bilayer, have been shown to retain the activity of incorporated H(+)-ATP synthases from chloroplasts in contrast to films prepared from the pure thiolipopeptide. The activity of the protein was demonstrated by impedance spectroscopy. The resistance decreased due to proton transport across the lipid film, which occurs as a consequence of adenosine triphosphate (ATP) hydrolysis. Several effects previously determined from kinetic measurements of the enzyme reconstituted in liposomes such as saturation with respect to the substrate (ATP), inhibition by venturicidin, activation by a positive potential pulse and increase of the proton current as a function of increasingly negative potentials have been confirmed also for this tethered membrane system. Changes in the impedance spectra due to the addition of ATP were fully reversible.
authors.affiliation: Max Planck Institute of Polymer Research, Ackermannweg 10, D-55128 Mainz, Germany. naumannr@mpip-mainz.mpg.de
authors.forename: R
authors.initials: R
authors.lastname: Naumann
authors.forename: T
authors.initials: T
authors.lastname: Baumgart
authors.forename: P
authors.initials: P
authors.lastname: Gräber
authors.forename: A
authors.initials: A
authors.lastname: Jonczyk
authors.forename: A
authors.initials: A
authors.lastname: Offenhäusser
authors.forename: W
authors.initials: W
authors.lastname: Knoll
mesh_terms: D002736:Chloroplasts / Q000201:enzymology*; D017097:Electric Impedance; D008051:Lipid Bilayers / Q000378:metabolism*; D008856:Microscopy, Fluorescence; D006180:Proton-Translocating ATPases / Q000378:metabolism*; D013057:Spectrum Analysis; D020349:Surface Plasmon Resonance
chemical_list: D008051:Lipid Bilayers; D006180:Proton-Translocating ATPases