Solid-state NMR chemical-shift perturbations indicate domain reorientation of the DnaG primase in the primosome of Helicobacter pylori. - Archive ouverte HAL Access content directly
Journal Articles Journal of Biomolecular NMR Year : 2016

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

Thomas Wiegand
Irina Gutsche

Abstract

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.
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Dates and versions

hal-01443306 , version 1 (24-11-2020)

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Carole Gardiennet, Thomas Wiegand, Alexandre Bazin, Riccardo Cadalbert, Britta Kunert, et al.. Solid-state NMR chemical-shift perturbations indicate domain reorientation of the DnaG primase in the primosome of Helicobacter pylori.. Journal of Biomolecular NMR, 2016, 64 (3), pp.189-95. ⟨10.1007/s10858-016-0018-0⟩. ⟨hal-01443306⟩
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