title: Solid-state NMR and EPR Spectroscopy of Mn2+-Substituted ATP-Fueled Protein Engines.
abstract: Paramagnetic metal ions deliver structural information both in EPR and solid-state NMR experiments, offering a profitable synergetic approach to study bio-macromolecules. We demonstrate the spectral consequences of Mg2+/ Mn2+substitution and the resulting information contents for two different ATP:Mg2+-fueled protein engines, a DnaB helicase from Helicobacter pylori active in the bacterial replisome, and the ABC transporter BmrA, a bacterial efflux pump. We show that, while EPR spectra report on metal binding and provide information on the geometry of the metal centers in the proteins, paramagnetic relaxation enhancements identified in the NMR spectra can be used to localize residues at the binding site. Protein engines are ubiquitous and the methods described herein should be applicable in a broad context.
authors.affiliation: Physical Chemistry, ETH Zurich, 8093, Zurich, Switzerland.
authors.forename: Thomas
authors.initials: T
authors.lastname: Wiegand
authors.affiliation: Molecular Microbiology and Structural Biochemistry, Labex Ecofect, UMR 5086 CNRS/Université de Lyon, 69367, Lyon, France.
authors.forename: Denis
authors.initials: D
authors.lastname: Lacabanne
authors.affiliation: Physical Chemistry, ETH Zurich, 8093, Zurich, Switzerland.
authors.forename: Katharina
authors.initials: K
authors.lastname: Keller
authors.affiliation: Physical Chemistry, ETH Zurich, 8093, Zurich, Switzerland.
authors.forename: Riccardo
authors.initials: R
authors.lastname: Cadalbert
authors.affiliation: Molecular Microbiology and Structural Biochemistry, Labex Ecofect, UMR 5086 CNRS/Université de Lyon, 69367, Lyon, France.
authors.forename: Lauriane
authors.initials: L
authors.lastname: Lecoq
authors.affiliation: Physical Chemistry, ETH Zurich, 8093, Zurich, Switzerland.
authors.forename: Maxim
authors.identifier: 0000-0003-3275-0714
authors.initials: M
authors.lastname: Yulikov
authors.affiliation: Molecular Microbiology and Structural Biochemistry, Labex Ecofect, UMR 5086 CNRS/Université de Lyon, 69367, Lyon, France.
authors.forename: Laurent
authors.initials: L
authors.lastname: Terradot
authors.affiliation: Physical Chemistry, ETH Zurich, 8093, Zurich, Switzerland.
authors.forename: Gunnar
authors.identifier: 0000-0001-6853-8585
authors.initials: G
authors.lastname: Jeschke
authors.affiliation: Physical Chemistry, ETH Zurich, 8093, Zurich, Switzerland.
authors.forename: Beat H
authors.identifier: 0000-0002-9107-4464
authors.initials: BH
authors.lastname: Meier
authors.affiliation: Molecular Microbiology and Structural Biochemistry, Labex Ecofect, UMR 5086 CNRS/Université de Lyon, 69367, Lyon, France.
authors.forename: Anja
authors.identifier: 0000-0001-8149-7941
authors.initials: A
authors.lastname: Böckmann
mesh_terms: D018528:ATP-Binding Cassette Transporters / Q000737:chemistry*; D000255:Adenosine Triphosphate / Q000737:chemistry*; D053488:DnaB Helicases / Q000737:chemistry*; D004578:Electron Spin Resonance Spectroscopy; D016480:Helicobacter pylori / Q000737:chemistry; D008274:Magnesium / Q000737:chemistry*; D008345:Manganese / Q000737:chemistry*; D008958:Models, Molecular; D019906:Nuclear Magnetic Resonance, Biomolecular
chemical_list: D018528:ATP-Binding Cassette Transporters; D008345:Manganese; D000255:Adenosine Triphosphate; D053488:DnaB Helicases; D008274:Magnesium
keywords: DNA replication; EPR spectroscopy; NMR spectroscopy; manganese; membrane proteins