Solid-state NMR chemical-shift perturbations indicate domain reorientation of the DnaG primase in the primosome of Helicobacter pylori
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. Mehr anzeigen
Persistenter Link
https://doi.org/10.3929/ethz-b-000114271Publikationsstatus
publishedExterne Links
Zeitschrift / Serie
Journal of Biomolecular NMRBand
Seiten / Artikelnummer
Verlag
SpringerThema
DnaB; DnaG; Primosome; Protein complexes; Sediments; Solid-state NMROrganisationseinheit
03496 - Meier, Beat H. (emeritus) / Meier, Beat H. (emeritus)
Förderung
159707 - NMR studies in the Solid State (SNF)
146757 - NMR studies in the Solid State (SNF)
Anmerkungen
It was possible to publish this article open access thanks to a Swiss National Licence with the publisher.