1. Solid-state NMR chemical-shift perturbations indicate domain reorientation of the DnaG primase in the primosome of Helicobacter pylori
- Author
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Anja Böckmann, Britta Kunert, Alexandre Bazin, Denis Lacabanne, Laurent Terradot, Irina Gutsche, Riccardo Cadalbert, Thomas Wiegand, Carole Gardiennet, Beat H. Meier, Cristallographie, Résonance Magnétique et Modélisations (CRM2), Université de Lorraine (UL)-Centre National de la Recherche Scientifique (CNRS), Microbiologie moléculaire et biochimie structurale / Molecular Microbiology and Structural Biochemistry (MMSB), Université Claude Bernard Lyon 1 (UCBL), Université de Lyon-Université de Lyon-Centre National de la Recherche Scientifique (CNRS), Physical Chemistry [ETH Zürich], Department of Chemistry and Applied Biosciences [ETH Zürich] (D-CHAB), Eidgenössische Technische Hochschule - Swiss Federal Institute of Technology [Zürich] (ETH Zürich)- Eidgenössische Technische Hochschule - Swiss Federal Institute of Technology [Zürich] (ETH Zürich), Institut de biologie structurale (IBS - UMR 5075 ), Centre National de la Recherche Scientifique (CNRS)-Université Grenoble Alpes [2016-2019] (UGA [2016-2019])-Institut de Recherche Interdisciplinaire de Grenoble (IRIG), Direction de Recherche Fondamentale (CEA) (DRF (CEA)), Commissariat à l'énergie atomique et aux énergies alternatives (CEA)-Commissariat à l'énergie atomique et aux énergies alternatives (CEA)-Direction de Recherche Fondamentale (CEA) (DRF (CEA)), Commissariat à l'énergie atomique et aux énergies alternatives (CEA)-Commissariat à l'énergie atomique et aux énergies alternatives (CEA), TGIR-RMN-THC FR3050, ANR-12-BS08-0013,XLproteinSSNMR,Etudes structurales du 75 kDa prion Sup35 : Vers la RMN du solide des protéines extra-larges(2012), Université Grenoble Alpes [2016-2019] (UGA [2016-2019])-Institut de Recherche Interdisciplinaire de Grenoble (IRIG), and Commissariat à l'énergie atomique et aux énergies alternatives (CEA)-Commissariat à l'énergie atomique et aux énergies alternatives (CEA)-Centre National de la Recherche Scientifique (CNRS)
- Subjects
0301 basic medicine ,Protein complexes ,DnaB ,Bazin ,DnaG ,01 natural sciences ,Biochemistry ,Solid-state NMR ,Primosome ,Sediments ,law.invention ,law ,MESH: Nuclear Magnetic Resonance, Biomolecular ,Crystallization ,Spectroscopy ,Irina ,[SDV.BBM.BS]Life Sciences [q-bio]/Biochemistry, Molecular Biology/Structural Biology [q-bio.BM] ,Chemistry ,Wiegand ,Riccardo ,3. Good health ,Solid-state nuclear magnetic resonance ,MESH: DNA Primase ,Denis ,MESH: Protein Domains ,Primase ,Böckmann ,Anja ,Carole ,Protein domain ,Terradot ,Laurent ,DNA Primase ,010402 general chemistry ,Domain (mathematical analysis) ,03 medical and health sciences ,Protein Domains ,Alexandre ,Beat H ,Nuclear Magnetic Resonance, Biomolecular ,dnaB helicase ,Gutsche ,Helicobacter pylori ,Gardiennet ,Cadalbert ,0104 chemical sciences ,Meier ,Crystallography ,Thomas ,030104 developmental biology ,Lacabanne ,Biophysics ,MESH: Helicobacter pylori ,Kunert ,Britta - Abstract
International audience; We here investigate the interactions between the DnaB helicase and the C-terminal domain of the corresponding DnaG primase of Helicobacter pylori using solid-state NMR. The difficult crystallization of this 387 kDa complex, where the two proteins interact in a six to three ratio, is circumvented by simple co-sedimentation of the two proteins directly into the MAS-NMR rotor. While the amount of information that can be extracted from such a large protein is still limited, we can assign a number of amino-acid residues experiencing significant chemical-shift perturbations upon helicase-primase complex formation. The location of these residues is used as a guide to model the interaction interface between the two proteins in the complex. Chemical-shift perturbations also reveal changes at the interaction interfaces of the hexameric HpDnaB assembly on HpDnaG binding. A structural model of the complex that explains the experimental findings is obtained.
- Published
- 2015
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