دورية أكاديمية
A real-time analysis of GFP unfolding by the AAA+ unfoldase PAN
| العنوان: | A real-time analysis of GFP unfolding by the AAA+ unfoldase PAN |
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| المؤلفون: | Krüger, Georg, Kirkpatrick, John, Mahieu, Emilie, Franzetti, Bruno, Gabel, Frank, Carlomagno, Teresa |
| المصدر: | Journal of Magnetic Resonance (JMR) 350 (2023) ; Journal of Magnetic Resonance (JMR) |
| بيانات النشر: | Elsevier |
| سنة النشر: | 2023 |
| المجموعة: | Institutional Repository of Leibniz Universität Hannover |
| مصطلحات موضوعية: | AAA+ ATPase, GFP, PAN, Protein unfolding, Real-time NMR spectroscopy, ddc:550, ddc:530 |
| الوصف: | Protein quality control systems are essential to maintain a healthy proteome. They often consist of an unfoldase unit, typically an AAA+ ATPase, coupled with a protease unit. In all kingdoms of life, they function to eliminate misfolded proteins, and thus prevent that their aggregates do harm to the cell, and to rapidly regulate protein levels in the presence of environmental changes. Despite the huge progress made in the past two decades in understanding the mechanism of function of protein degradation systems, the fate of the substrate during the unfolding and proteolytic processes remains poorly understood. Here we exploit an NMR-based approach to monitor GFP processing by the archaeal PAN unfoldase and the PAN–20S degradation system in real time. We find that PAN-dependent unfolding of GFP does not involve the release of partially-folded GFP molecules resulting from futile unfolding attempts. In contrast, once stably engaged with PAN, GFP molecules are efficiently transferred to the proteolytic chamber of the 20S subunit, despite the only weak affinity of PAN for the 20S subunit in the absence of substrate. This is essential to guarantee that unfolded but not proteolyzed proteins are not released into solution, where they would form toxic aggregates. The results of our studies are in good agreement with previous results derived from real-time small-angle-neutron-scattering experiments and have the advantage of allowing the investigation of substrates and products at amino-acid resolution. |
| نوع الوثيقة: | article in journal/newspaper |
| اللغة: | English |
| تدمد: | 1090-7807 |
| العلاقة: | ESSN:1096-0856; http://dx.doi.org/10.15488/14997Test; https://www.repo.uni-hannover.de/handle/123456789/15116Test |
| DOI: | 10.15488/14997 |
| الإتاحة: | https://doi.org/10.15488/14997Test https://doi.org/10.1016/j.jmr.2023.107431Test https://www.repo.uni-hannover.de/handle/123456789/15116Test |
| حقوق: | CC BY-NC-ND 4.0 Unported ; https://creativecommons.org/licenses/by-nc-nd/4.0Test/ ; frei zugänglich |
| رقم الانضمام: | edsbas.5E0CD601 |
| قاعدة البيانات: | BASE |
| تدمد: | 10907807 |
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| DOI: | 10.15488/14997 |