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Structural and functional insights into the inhibition of human voltage-gated sodium channels by μ-conotoxin KIIIA disulfide isomers
μ-Conotoxins are components of cone snail venom, well-known for their analgesic activity through potent inhibition of voltage-gated sodium channel (Na(V)) subtypes, including Na(V)1.7. These small, disulfide-rich peptides are typically stabilized by three disulfide bonds arranged in a ‘native’ CysI-...
Autores principales: | , , , , |
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Formato: | Online Artículo Texto |
Lenguaje: | English |
Publicado: |
American Society for Biochemistry and Molecular Biology
2022
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Materias: | |
Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC8927997/ https://www.ncbi.nlm.nih.gov/pubmed/35167877 http://dx.doi.org/10.1016/j.jbc.2022.101728 |
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author | Tran, Hue N.T. McMahon, Kirsten L. Deuis, Jennifer R. Vetter, Irina Schroeder, Christina I. |
author_facet | Tran, Hue N.T. McMahon, Kirsten L. Deuis, Jennifer R. Vetter, Irina Schroeder, Christina I. |
author_sort | Tran, Hue N.T. |
collection | PubMed |
description | μ-Conotoxins are components of cone snail venom, well-known for their analgesic activity through potent inhibition of voltage-gated sodium channel (Na(V)) subtypes, including Na(V)1.7. These small, disulfide-rich peptides are typically stabilized by three disulfide bonds arranged in a ‘native’ CysI-CysIV, CysII-CysV, CysIII-CysVI pattern of disulfide connectivity. However, μ-conotoxin KIIIA, the smallest and most studied μ-conotoxin with inhibitory activity at Na(V)1.7, forms two distinct disulfide bond isomers during thermodynamic oxidative folding, including Isomer 1 (CysI-CysV, CysII-CysIV, CysIII-CysVI) and Isomer 2 (CysI-CysVI, CysII-CysIV, CysIII-CysV), but not the native μ-conotoxin arrangement. To date, there has been no study on the structure and activity of KIIIA comprising the native μ-conotoxin disulfide bond arrangement. Here, we evaluated the synthesis, potency, sodium channel subtype selectivity, and 3D structure of the three isomers of KIIIA. Using a regioselective disulfide bond-forming strategy, we synthetically produced the three μ-conotoxin KIIIA isomers displaying distinct bioactivity and Na(V) subtype selectivity across human Na(V) channel subtypes 1.2, 1.4, and 1.7. We show that Isomer 1 inhibits Na(V) subtypes with a rank order of potency of Na(V)1.4 > 1.2 > 1.7 and Isomer 2 in the order of Na(V)1.4≈1.2 > 1.7, while the native isomer inhibited Na(V)1.4 > 1.7≈1.2. The three KIIIA isomers were further evaluated by NMR solution structure analysis and molecular docking with hNa(V)1.2. Our study highlights the importance of investigating alternate disulfide isomers, as disulfide connectivity affects not only the overall structure of the peptides but also the potency and subtype selectivity of μ-conotoxins targeting therapeutically relevant Na(V) subtypes. |
format | Online Article Text |
id | pubmed-8927997 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2022 |
publisher | American Society for Biochemistry and Molecular Biology |
record_format | MEDLINE/PubMed |
spelling | pubmed-89279972022-03-24 Structural and functional insights into the inhibition of human voltage-gated sodium channels by μ-conotoxin KIIIA disulfide isomers Tran, Hue N.T. McMahon, Kirsten L. Deuis, Jennifer R. Vetter, Irina Schroeder, Christina I. J Biol Chem Research Article μ-Conotoxins are components of cone snail venom, well-known for their analgesic activity through potent inhibition of voltage-gated sodium channel (Na(V)) subtypes, including Na(V)1.7. These small, disulfide-rich peptides are typically stabilized by three disulfide bonds arranged in a ‘native’ CysI-CysIV, CysII-CysV, CysIII-CysVI pattern of disulfide connectivity. However, μ-conotoxin KIIIA, the smallest and most studied μ-conotoxin with inhibitory activity at Na(V)1.7, forms two distinct disulfide bond isomers during thermodynamic oxidative folding, including Isomer 1 (CysI-CysV, CysII-CysIV, CysIII-CysVI) and Isomer 2 (CysI-CysVI, CysII-CysIV, CysIII-CysV), but not the native μ-conotoxin arrangement. To date, there has been no study on the structure and activity of KIIIA comprising the native μ-conotoxin disulfide bond arrangement. Here, we evaluated the synthesis, potency, sodium channel subtype selectivity, and 3D structure of the three isomers of KIIIA. Using a regioselective disulfide bond-forming strategy, we synthetically produced the three μ-conotoxin KIIIA isomers displaying distinct bioactivity and Na(V) subtype selectivity across human Na(V) channel subtypes 1.2, 1.4, and 1.7. We show that Isomer 1 inhibits Na(V) subtypes with a rank order of potency of Na(V)1.4 > 1.2 > 1.7 and Isomer 2 in the order of Na(V)1.4≈1.2 > 1.7, while the native isomer inhibited Na(V)1.4 > 1.7≈1.2. The three KIIIA isomers were further evaluated by NMR solution structure analysis and molecular docking with hNa(V)1.2. Our study highlights the importance of investigating alternate disulfide isomers, as disulfide connectivity affects not only the overall structure of the peptides but also the potency and subtype selectivity of μ-conotoxins targeting therapeutically relevant Na(V) subtypes. American Society for Biochemistry and Molecular Biology 2022-02-12 /pmc/articles/PMC8927997/ /pubmed/35167877 http://dx.doi.org/10.1016/j.jbc.2022.101728 Text en © 2022 The Authors https://creativecommons.org/licenses/by-nc-nd/4.0/This is an open access article under the CC BY-NC-ND license (http://creativecommons.org/licenses/by-nc-nd/4.0/). |
spellingShingle | Research Article Tran, Hue N.T. McMahon, Kirsten L. Deuis, Jennifer R. Vetter, Irina Schroeder, Christina I. Structural and functional insights into the inhibition of human voltage-gated sodium channels by μ-conotoxin KIIIA disulfide isomers |
title | Structural and functional insights into the inhibition of human voltage-gated sodium channels by μ-conotoxin KIIIA disulfide isomers |
title_full | Structural and functional insights into the inhibition of human voltage-gated sodium channels by μ-conotoxin KIIIA disulfide isomers |
title_fullStr | Structural and functional insights into the inhibition of human voltage-gated sodium channels by μ-conotoxin KIIIA disulfide isomers |
title_full_unstemmed | Structural and functional insights into the inhibition of human voltage-gated sodium channels by μ-conotoxin KIIIA disulfide isomers |
title_short | Structural and functional insights into the inhibition of human voltage-gated sodium channels by μ-conotoxin KIIIA disulfide isomers |
title_sort | structural and functional insights into the inhibition of human voltage-gated sodium channels by μ-conotoxin kiiia disulfide isomers |
topic | Research Article |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC8927997/ https://www.ncbi.nlm.nih.gov/pubmed/35167877 http://dx.doi.org/10.1016/j.jbc.2022.101728 |
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