Solid-state NMR and EPR Spectroscopy of Mn2+ -Substituted ATP-Fueled Protein Engines.
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.
['Wiegand|Thomas|T|', 'Lacabanne|Denis|D|', 'Keller|Katharina|K|', 'Cadalbert|Riccardo|R|', 'Lecoq|Lauriane|L|', 'Yulikov|Maxim|M|0000-0003-3275-0714', 'Terradot|Laurent|L|', 'Jeschke|Gunnar|G|0000-0001-6853-8585', 'Meier|Beat H|BH|0000-0002-9107-4464', 'Böckmann|Anja|A|0000-0001-8149-7941']
D018528:ATP-Binding Cassette Transporters D000255:Adenosine Triphosphate D053488:DnaB Helicases D004578:Electron Spin Resonance Spectroscopy D016480:Helicobacter pylori D008274:Magnesium D008345:Manganese D008958:Models, Molecular D019906:Nuclear Magnetic Resonance, Biomolecular
EPR spectroscopy DNA replication NMR spectroscopy manganese membrane proteins