PROTstretch - Dynamic structure of a nanomachine involved in proteome quality control: a combined NMR/SAXS/SANS study of the PAN unfoldase
Biochemistry
Final Report Abstract
We have addressed all the three aspects that we had proposed to address in the project. To the first aspect, we have found that the symmetry of PAN in solution cannot be reconciled with the spiral staircase model, indicating the importance of NMR data to verify high-resolution data acquired in “solid” matrices. Our data allow us to propose a revised mechanism for the function of AAA+ ATPases which has the merit of reconciling all biochemical and biophysical data and also be in agreement with minor conformations observed by electron microscopy. To the second aspect, we have been unable to characterize an encounter complex by NMR because of the fast hydrolysis of ATP. However, we have demonstrated that neither the OB nor the CC domains are responsible for substrate recruitment. To the third aspect, we have demonstrated that GFP is unfolded without release of partially unfolded intermediates in solution. We have also demonstrated that the PAN- catalysed denaturation reaction is tightly coupled to degradation by the 20S protease, thus avoiding the formation of dangerous aggregates in the cells.
Publications
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Observing Protein Degradation by the PAN-20S Proteasome by Time-Resolved Neutron Scattering. Biophysical Journal, 119(2), 375-388.
Mahieu, Emilie; Covès, Jacques; Krüger, Georg; Martel, Anne; Moulin, Martine; Carl, Nico; Härtlein, Michael; Carlomagno, Teresa; Franzetti, Bruno & Gabel, Frank
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An NMR Study of a 300-kDa AAA+ Unfoldase. Journal of Molecular Biology, 435(11), 167997.
Krüger, Georg; Kirkpatrick, John; Mahieu, Emilie; Franzetti, Bruno; Gabel, Frank & Carlomagno, Teresa
