Solid-state NMR chemical-shift perturbations indicate domain reorientation of the DnaG primase in the primosome of Helicobacter pylori

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• 作者：Carole Gardiennet ; Thomas Wiegand ; Alexandre Bazin…
• 关键词：Solid ; state NMR ; Sediments ; Protein complexes ; DnaB ; DnaG ; Primosome
• 刊名：Journal of Biomolecular NMR
• 出版年：2016
• 出版时间：March 2016
• 年：2016
• 卷：64
• 期：3
• 页码：189-195
• 全文大小：5,549 KB
• 参考文献：Abdul Rehman SA, Verma V, Mazumder M et al (2013) Crystal structure and mode of helicase binding of the C-terminal domain of primase from Helicobacter pylori. J Bacteriol 195:2826–2838. doi:10.​1128/​JB.​00091-13 CrossRef
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• 作者单位：Carole Gardiennet (1) (4)
  Thomas Wiegand (2)
  Alexandre Bazin (1)
  Riccardo Cadalbert (2)
  Britta Kunert (1)
  Denis Lacabanne (1)
  Irina Gutsche (3)
  Laurent Terradot (1)
  Beat H. Meier (2)
  Anja Böckmann (1)
  
  1. Molecular Microbiology and Structural Biochemistry, Labex Ecofect, UMR 5086 CNRS, Université de Lyon 1, 7 passage du Vercors, 69007, Lyon, France
  4. CNRS, CRM2, UMR 7036, Université de Lorraine, 54506, Vandoeuvre-lès-Nancy, France
  2. Physical Chemistry, ETH Zurich, Vladimir-Prelog-Weg 2, 8093, Zurich, Switzerland
  3. Institut de Biologie Structurale (IBS), CNRS, IBS; CEA, IBS, Université Grenoble Alpes, 38044, Grenoble, France
  
• 刊物类别：Physics and Astronomy
• 刊物主题：Physics
  Biophysics and Biomedical Physics
  Polymer Sciences
  Biochemistry
  
• 出版者：Springer Netherlands
• ISSN：1573-5001

文摘

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.