Characterization of clostridium botulinum neurotoxin serotype A (BoNT/A) and fibroblast growth factor receptor interactions using novel receptor dimerization assay
| Autor: | Ron S. Broide, Birgitte S. Jacky, Amy Brideau-Andersen, Catherine Rheaume, David M. Jameson, Nicholas G. James, Bethany J. Sanstrum, Shiazah Z. Malik |
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| Jazyk: | angličtina |
| Rok vydání: | 2021 |
| Předmět: |
0301 basic medicine
Cell biology Molecular biology media_common.quotation_subject Science Neurotoxins Nerve Tissue Proteins Receptors Cell Surface medicine.disease_cause Serogroup Transfection PC12 Cells Article 03 medical and health sciences 0302 clinical medicine Protein Domains Cell surface receptor Ganglioside binding Gangliosides medicine Clostridium botulinum Neurotoxin Animals Botulinum Toxins Type A Internalization Receptor media_common Neurons Multidisciplinary Binding Sites Membrane Glycoproteins Chemistry Cell Membrane Receptors Fibroblast Growth Factor Rats ErbB Receptors 030104 developmental biology Fibroblast growth factor receptor Medicine Dimerization 030217 neurology & neurosurgery Binding domain Protein Binding Signal Transduction Neuroscience |
| Zdroj: | Scientific Reports, Vol 11, Iss 1, Pp 1-12 (2021) Scientific Reports |
| ISSN: | 2045-2322 |
| Popis: | Clostridium botulinum neurotoxin serotype A (BoNT/A) is a potent neurotoxin that serves as an effective therapeutic for several neuromuscular disorders via induction of temporary muscular paralysis. Specific binding and internalization of BoNT/A into neuronal cells is mediated by its binding domain (HC/A), which binds to gangliosides, including GT1b, and protein cell surface receptors, including SV2. Previously, recombinant HC/A was also shown to bind to FGFR3. As FGFR dimerization is an indirect measure of ligand-receptor binding, an FCS & TIRF receptor dimerization assay was developed to measure rHC/A-induced dimerization of fluorescently tagged FGFR subtypes (FGFR1-3) in cells. rHC/A dimerized FGFR subtypes in the rank order FGFR3c (EC50 ≈ 27 nM) > FGFR2b (EC50 ≈ 70 nM) > FGFR1c (EC50 ≈ 163 nM); rHC/A dimerized FGFR3c with similar potency as the native FGFR3c ligand, FGF9 (EC50 ≈ 18 nM). Mutating the ganglioside binding site in HC/A, or removal of GT1b from the media, resulted in decreased dimerization. Interestingly, reduced dimerization was also observed with an SV2 mutant variant of HC/A. Overall, the results suggest that the FCS & TIRF receptor dimerization assay can assess FGFR dimerization with known and novel ligands and support a model wherein HC/A, either directly or indirectly, interacts with FGFRs and induces receptor dimerization. |
| Databáze: | OpenAIRE |
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