Botulinum Neurotoxin A4 Has a 1000-Fold Reduced Potency Due to Three Single Amino Acid Alterations in the Protein Receptor Binding Domain
Botulinum neurotoxin subtype A4 (BoNT/A4) is ~1000-fold less potent than BoNT/A1. This study addresses the basis for low BoNT/A4 potency. Utilizing BoNT/A1-A4 and BoNT/A4-A1 Light Chain-Heavy Chain (LC-HC) chimeras, HC-A4 was responsible for low BoNT/A4 potency. Earlier studies showed BoNT/A1-recept...
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MDPI AG
2023-03-01
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| Series: | International Journal of Molecular Sciences |
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| Online Access: | https://www.mdpi.com/1422-0067/24/6/5690 |
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| author | William H. Tepp Marite Bradshaw Alexander P. Gardner Rebecca L. Kaufman Joseph T. Barbieri Sabine Pellett |
| author_facet | William H. Tepp Marite Bradshaw Alexander P. Gardner Rebecca L. Kaufman Joseph T. Barbieri Sabine Pellett |
| author_sort | William H. Tepp |
| collection | DOAJ |
| description | Botulinum neurotoxin subtype A4 (BoNT/A4) is ~1000-fold less potent than BoNT/A1. This study addresses the basis for low BoNT/A4 potency. Utilizing BoNT/A1-A4 and BoNT/A4-A1 Light Chain-Heavy Chain (LC-HC) chimeras, HC-A4 was responsible for low BoNT/A4 potency. Earlier studies showed BoNT/A1-receptor binding domain (Hcc) bound a β-strand peptide (556–564) and glycan-N<sup>559</sup> within Luminal Domain 4 (LD4) of SV2C, the BoNT/A protein receptor. Relative to BoNT/A1, the Hcc of BoNT/A4 possesses two amino acid variants (D<sup>1141</sup> and N<sup>1142</sup>) within the β-peptide binding interface and one amino acid variant (R<sup>1292</sup>) located near the SV2C glycan-N<sup>559</sup>. Introduction of BoNT/A4 β-strand peptide variant (D<sup>1141</sup> and N<sup>1142</sup>) into BoNT/A1 reduced toxin potency 30-fold, and additional introduction of the BoNT/A4 glycan-N<sup>559</sup> variant (D<sup>1141</sup>, N<sup>1142</sup>, and R<sup>1292</sup>) further reduced toxin potency to approach BoNT/A4. While introduction of BoNT/A1 glycan-N<sup>559</sup> variant (G<sup>1292</sup>) into BoNT/A4 did not alter toxin potency, additional introduction of BoNT/A1 β-strand peptide variants (G<sup>1141</sup>, S<sup>1142</sup>, and G<sup>1292</sup>) resulted in potency approaching BoNT/A1 potency. Thus, outcomes from these functional and modeling studies indicate that in rodent models, disruption of Hcc -SV2C β-peptide and -glycan-N<sup>559</sup> interactions mediate low BoNT/A4 potency, while in human motor neurons, disruption of Hcc-SV2C β-peptide alone mediates low BoNT/A4 potency, which link to a species-specific variation at SV2C<sup>563</sup>. |
| first_indexed | 2024-03-11T06:25:08Z |
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| language | English |
| last_indexed | 2024-03-11T06:25:08Z |
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| spelling | doaj.art-874c916fba1b4549a67274fe8a61887c2023-11-17T11:37:37ZengMDPI AGInternational Journal of Molecular Sciences1661-65961422-00672023-03-01246569010.3390/ijms24065690Botulinum Neurotoxin A4 Has a 1000-Fold Reduced Potency Due to Three Single Amino Acid Alterations in the Protein Receptor Binding DomainWilliam H. Tepp0Marite Bradshaw1Alexander P. Gardner2Rebecca L. Kaufman3Joseph T. Barbieri4Sabine Pellett5Department of Bacteriology, University of Wisconsin, Madison, WI 53706, USADepartment of Bacteriology, University of Wisconsin, Madison, WI 53706, USADepartment of Microbiology and Molecular Genetics, Medical College of Wisconsin, Milwaukee, WI 53226, USADepartment of Bacteriology, University of Wisconsin, Madison, WI 53706, USADepartment of Microbiology and Molecular Genetics, Medical College of Wisconsin, Milwaukee, WI 53226, USADepartment of Bacteriology, University of Wisconsin, Madison, WI 53706, USABotulinum neurotoxin subtype A4 (BoNT/A4) is ~1000-fold less potent than BoNT/A1. This study addresses the basis for low BoNT/A4 potency. Utilizing BoNT/A1-A4 and BoNT/A4-A1 Light Chain-Heavy Chain (LC-HC) chimeras, HC-A4 was responsible for low BoNT/A4 potency. Earlier studies showed BoNT/A1-receptor binding domain (Hcc) bound a β-strand peptide (556–564) and glycan-N<sup>559</sup> within Luminal Domain 4 (LD4) of SV2C, the BoNT/A protein receptor. Relative to BoNT/A1, the Hcc of BoNT/A4 possesses two amino acid variants (D<sup>1141</sup> and N<sup>1142</sup>) within the β-peptide binding interface and one amino acid variant (R<sup>1292</sup>) located near the SV2C glycan-N<sup>559</sup>. Introduction of BoNT/A4 β-strand peptide variant (D<sup>1141</sup> and N<sup>1142</sup>) into BoNT/A1 reduced toxin potency 30-fold, and additional introduction of the BoNT/A4 glycan-N<sup>559</sup> variant (D<sup>1141</sup>, N<sup>1142</sup>, and R<sup>1292</sup>) further reduced toxin potency to approach BoNT/A4. While introduction of BoNT/A1 glycan-N<sup>559</sup> variant (G<sup>1292</sup>) into BoNT/A4 did not alter toxin potency, additional introduction of BoNT/A1 β-strand peptide variants (G<sup>1141</sup>, S<sup>1142</sup>, and G<sup>1292</sup>) resulted in potency approaching BoNT/A1 potency. Thus, outcomes from these functional and modeling studies indicate that in rodent models, disruption of Hcc -SV2C β-peptide and -glycan-N<sup>559</sup> interactions mediate low BoNT/A4 potency, while in human motor neurons, disruption of Hcc-SV2C β-peptide alone mediates low BoNT/A4 potency, which link to a species-specific variation at SV2C<sup>563</sup>.https://www.mdpi.com/1422-0067/24/6/5690botulinum neurotoxinclostridium botulinumSV2CreceptorpotencyBoNT/A4 |
| spellingShingle | William H. Tepp Marite Bradshaw Alexander P. Gardner Rebecca L. Kaufman Joseph T. Barbieri Sabine Pellett Botulinum Neurotoxin A4 Has a 1000-Fold Reduced Potency Due to Three Single Amino Acid Alterations in the Protein Receptor Binding Domain International Journal of Molecular Sciences botulinum neurotoxin clostridium botulinum SV2C receptor potency BoNT/A4 |
| title | Botulinum Neurotoxin A4 Has a 1000-Fold Reduced Potency Due to Three Single Amino Acid Alterations in the Protein Receptor Binding Domain |
| title_full | Botulinum Neurotoxin A4 Has a 1000-Fold Reduced Potency Due to Three Single Amino Acid Alterations in the Protein Receptor Binding Domain |
| title_fullStr | Botulinum Neurotoxin A4 Has a 1000-Fold Reduced Potency Due to Three Single Amino Acid Alterations in the Protein Receptor Binding Domain |
| title_full_unstemmed | Botulinum Neurotoxin A4 Has a 1000-Fold Reduced Potency Due to Three Single Amino Acid Alterations in the Protein Receptor Binding Domain |
| title_short | Botulinum Neurotoxin A4 Has a 1000-Fold Reduced Potency Due to Three Single Amino Acid Alterations in the Protein Receptor Binding Domain |
| title_sort | botulinum neurotoxin a4 has a 1000 fold reduced potency due to three single amino acid alterations in the protein receptor binding domain |
| topic | botulinum neurotoxin clostridium botulinum SV2C receptor potency BoNT/A4 |
| url | https://www.mdpi.com/1422-0067/24/6/5690 |
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