Academic Journal
Pulsed EPR Methods in the Angstrom to Nanometre Scale Shed Light on the Conformational Flexibility of a Fluoride Riboswitch
| العنوان: | Pulsed EPR Methods in the Angstrom to Nanometre Scale Shed Light on the Conformational Flexibility of a Fluoride Riboswitch |
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| المؤلفون: | Remmel, Laura, Meyer, Andreas, Ackermann, Katrin, Hagelueken, Gregor, Bennati, Marina, Bode, Bela E. |
| المساهمون: | Engineering and Physical Sciences Research Council, Biotechnology and Biological Sciences Research Council, HORIZON EUROPE European Research Council, Deutsche Forschungsgemeinschaft, Max-Planck-Gesellschaft, Royal Society |
| المصدر: | Angewandte Chemie International Edition ; volume 63, issue 49 ; ISSN 1433-7851 1521-3773 |
| بيانات النشر: | Wiley |
| سنة النشر: | 2024 |
| المجموعة: | Wiley Online Library (Open Access Articles via Crossref) |
| الوصف: | Riboswitches control gene regulation upon external stimuli such as environmental factors or ligand binding. The fluoride sensing riboswitch from Thermotoga petrophila is a complex regulatory RNA proposed to be involved in resistance to F − cytotoxicity. The details of structure and dynamics underpinning the regulatory mechanism are currently debated. Here we demonstrate that a combination of pulsed electron paramagnetic resonance (ESR/EPR) spectroscopies, detecting distances in the angstrom to nanometre range, can probe distinct regions of conformational flexibility in this riboswitch. PELDOR (pulsed electron‐electron double resonance) revealed a similar preorganisation of the sensing domain in three forms, i.e. the free aptamer, the Mg 2+ ‐bound apo , and the F − ‐bound holo form. 19 F ENDOR (electron‐nuclear double resonance) was used to investigate the active site structure of the F − ‐bound holo form. Distance distributions without a priori structural information were compared with in silico modelling of spin label conformations based on the crystal structure. While PELDOR, probing the periphery of the RNA fold, revealed conformational flexibility of the RNA backbone, ENDOR indicated low structural heterogeneity at the ligand binding site. Overall, the combination of PELDOR and ENDOR with sub‐angstrom precision gave insight into structural organisation and flexibility of a riboswitch, not easily attainable by other biophysical techniques. |
| نوع الوثيقة: | article in journal/newspaper |
| اللغة: | English |
| DOI: | 10.1002/anie.202411241 |
| الاتاحة: | http://dx.doi.org/10.1002/anie.202411241 https://onlinelibrary.wiley.com/doi/pdf/10.1002/anie.202411241 |
| Rights: | http://creativecommons.org/licenses/by/4.0/ |
| رقم الانضمام: | edsbas.27DBCB1F |
| قاعدة البيانات: | BASE |
| DOI: | 10.1002/anie.202411241 |
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