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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-...

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Autores principales: Tran, Hue N.T., McMahon, Kirsten L., Deuis, Jennifer R., Vetter, Irina, Schroeder, Christina I.
Formato: Online Artículo Texto
Lenguaje:English
Publicado: American Society for Biochemistry and Molecular Biology 2022
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.
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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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