التفاصيل البيبلوغرافية
| العنوان: |
Investigation of the Fuzzy Complex between RSV Nucleoprotein and Phosphoprotein to Optimize an Inhibition Assay by Fluorescence Polarization |
| المؤلفون: |
Silva Khodjoyan, Deborha Morissette, Fortune Hontonnou, Luis Checa Ruano, Charles-Adrien Richard, Olivier Sperandio, Jean-François Eléouët, Marie Galloux, Philippe Durand, Stéphanie Deville-Foillard, Christina Sizun |
| المصدر: |
International Journal of Molecular Sciences; Volume 24; Issue 1; Pages: 569 |
| بيانات النشر: |
Multidisciplinary Digital Publishing Institute |
| سنة النشر: |
2022 |
| المجموعة: |
MDPI Open Access Publishing |
| مصطلحات موضوعية: |
respiratory syncytial virus, phosphoprotein, nucleoprotein, replication complex, fuzzy complex, protein–protein interaction, PPI inhibition, nuclear magnetic resonance, fluorescence anisotropy, B-cyano BODIPYs |
| جغرافية الموضوع: |
agris |
| الوصف: |
The interaction between Respiratory Syncytial Virus phosphoprotein P and nucleoprotein N is essential for the formation of the holo RSV polymerase that carries out replication. In vitro screening of antivirals targeting the N-P protein interaction requires a molecular interaction model, ideally consisting of a complex between N protein and a short peptide corresponding to the C-terminal tail of the P protein. However, the flexibility of C-terminal P peptides as well as their phosphorylation status play a role in binding and may bias the outcome of an inhibition assay. We therefore investigated binding affinities and dynamics of this interaction by testing two N protein constructs and P peptides of different lengths and composition, using nuclear magnetic resonance and fluorescence polarization (FP). We show that, although the last C-terminal Phe241 residue is the main determinant for anchoring P to N, only longer peptides afford sub-micromolar affinity, despite increasing mobility towards the N-terminus. We investigated competitive binding by peptides and small compounds, including molecules used as fluorescent labels in FP. Based on these results, we draw optimized parameters for a robust RSV N-P inhibition assay and validated this assay with the M76 molecule, which displays antiviral properties, for further screening of chemical libraries. |
| نوع الوثيقة: |
text |
| وصف الملف: |
application/pdf |
| اللغة: |
English |
| Relation: |
Macromolecules; https://dx.doi.org/10.3390/ijms24010569 |
| DOI: |
10.3390/ijms24010569 |
| الاتاحة: |
https://doi.org/10.3390/ijms24010569 |
| Rights: |
https://creativecommons.org/licenses/by/4.0/ |
| رقم الانضمام: |
edsbas.5D4FFC7C |
| قاعدة البيانات: |
BASE |