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标题：Characterization of clostridium botulinum neurotoxin serotype A (BoNT/A) and fibroblast growth factor receptor interactions using novel receptor dimerization assay
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作者：Nicholas G. James ; Shiazah Malik ; Bethany J. Sanstrum 等
期刊名称：Scientific Reports
电子版ISSN：2045-2322
出版年度：2021
卷号：11
DOI：10.1038/s41598-021-87331-7
语种：English
出版社：Springer Nature
摘要：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 (H C/A), which binds to gangliosides, including GT1b, and protein cell surface receptors, including SV2. Previously, recombinant H C/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 rH C/A-induced dimerization of fluorescently tagged FGFR subtypes (FGFR1-3) in cells. rH C/A dimerized FGFR subtypes in the rank order FGFR3c (EC 50 ≈ 27 nM) > FGFR2b (EC 50 ≈ 70 nM) > FGFR1c (EC 50 ≈ 163 nM); rH C/A dimerized FGFR3c with similar potency as the native FGFR3c ligand, FGF9 (EC 50 ≈ 18 nM). Mutating the ganglioside binding site in H C/A, or removal of GT1b from the media, resulted in decreased dimerization. Interestingly, reduced dimerization was also observed with an SV2 mutant variant of H C/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 H C/A, either directly or indirectly, interacts with FGFRs and induces receptor dimerization.