Details
Original language | English |
---|---|
Article number | 167997 |
Journal | Journal of molecular biology |
Volume | 435 |
Issue number | 11 |
Publication status | Published - 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
- Biochemistry, Genetics and Molecular Biology(all)
- Biophysics
- Biochemistry, Genetics and Molecular Biology(all)
- Structural Biology
- Biochemistry, Genetics and Molecular Biology(all)
- Molecular Biology
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In: Journal of molecular biology, Vol. 435, No. 11, 167997, 01.06.2023.
Research output: Contribution to journal › Article › Research › peer review
}
TY - JOUR
T1 - An NMR Study of a 300-kDa AAA+ Unfoldase
AU - Krüger, Georg
AU - Kirkpatrick, John
AU - Mahieu, Emilie
AU - Franzetti, Bruno
AU - Gabel, Frank
AU - Carlomagno, Teresa
N1 - Funding Information: This work was funded by the Deutsche Forschungsgemeinschaft through grant CA294/13-1 to T.C and by the Leverhulme Trust through a Leverhulme International Professorship to T.C. We thank Susanne zur Lage (HZI Braunschweig) for help with sample preparation.
PY - 2023/6/1
Y1 - 2023/6/1
N2 - 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.
AB - 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.
KW - AAA+ ATPase
KW - methyl NMR
KW - PAN
KW - unfoldase
UR - http://www.scopus.com/inward/record.url?scp=85165710380&partnerID=8YFLogxK
U2 - 10.1016/j.jmb.2023.167997
DO - 10.1016/j.jmb.2023.167997
M3 - Article
C2 - 37330287
AN - SCOPUS:85165710380
VL - 435
JO - Journal of molecular biology
JF - Journal of molecular biology
SN - 0022-2836
IS - 11
M1 - 167997
ER -