An NMR Study of a 300-kDa AAA+ Unfoldase

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Authors

  • Georg Krüger
  • John Kirkpatrick
  • Emilie Mahieu
  • Bruno Franzetti
  • Frank Gabel
  • Teresa Carlomagno

External Research Organisations

  • University of Birmingham
  • University Grenoble-Alpes (UGA)
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Details

Original languageEnglish
Article number167997
JournalJournal of molecular biology
Volume435
Issue number11
Publication statusPublished - 1 Jun 2023

Abstract

AAA+ ATPases are ubiquitous hexameric unfoldases acting in cellular protein quality control. In complex with proteases, they form protein degradation machinery (the proteasome) in both archaea and eukaryotes. Here, we use solution-state NMR spectroscopy to determine the symmetry properties of the archaeal PAN AAA+ unfoldase and gain insights into its functional mechanism. PAN consists of three folded domains: the coiled-coil (CC), OB and ATPase domains. We find that full-length PAN assembles into a hexamer with C2 symmetry, and that this symmetry extends over the CC, OB and ATPase domains. The NMR data, collected in the absence of substrate, are incompatible with the spiral staircase structure observed in electron-microscopy studies of archaeal PAN in the presence of substrate and in electron-microscopy studies of eukaryotic unfoldases both in the presence and in the absence of substrate. Based on the C2 symmetry revealed by NMR spectroscopy in solution, we propose that archaeal ATPases are flexible enzymes, which can adopt distinct conformations in different conditions. This study reaffirms the importance of studying dynamic systems in solution.

Keywords

    AAA+ ATPase, methyl NMR, PAN, unfoldase

ASJC Scopus subject areas

Cite this

An NMR Study of a 300-kDa AAA+ Unfoldase. / Krüger, Georg; Kirkpatrick, John; Mahieu, Emilie et al.
In: Journal of molecular biology, Vol. 435, No. 11, 167997, 01.06.2023.

Research output: Contribution to journalArticleResearchpeer review

Krüger, G, Kirkpatrick, J, Mahieu, E, Franzetti, B, Gabel, F & Carlomagno, T 2023, 'An NMR Study of a 300-kDa AAA+ Unfoldase', Journal of molecular biology, vol. 435, no. 11, 167997. https://doi.org/10.1016/j.jmb.2023.167997
Krüger, G., Kirkpatrick, J., Mahieu, E., Franzetti, B., Gabel, F., & Carlomagno, T. (2023). An NMR Study of a 300-kDa AAA+ Unfoldase. Journal of molecular biology, 435(11), Article 167997. https://doi.org/10.1016/j.jmb.2023.167997
Krüger G, Kirkpatrick J, Mahieu E, Franzetti B, Gabel F, Carlomagno T. An NMR Study of a 300-kDa AAA+ Unfoldase. Journal of molecular biology. 2023 Jun 1;435(11):167997. doi: 10.1016/j.jmb.2023.167997
Krüger, Georg ; Kirkpatrick, John ; Mahieu, Emilie et al. / An NMR Study of a 300-kDa AAA+ Unfoldase. In: Journal of molecular biology. 2023 ; Vol. 435, No. 11.
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