O1-conotoxin Tx6.7 cloned from the genomic DNA of Conus textile that inhibits calcium currents
Abstract Background: Conotoxins exhibit great potential as neuropharmacology tools and therapeutic candidates due to their high affinity and specificity for ion channels, neurotransmitter receptors or transporters. The traditional methods to discover new conotoxins are peptide purification from the...
Published in: | Journal of Venomous Animals and Toxins including Tropical Diseases |
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ftdoajarticles:oai:doaj.org/article:005360dd729644999e601e2008bca650 2023-06-11T04:09:39+02:00 O1-conotoxin Tx6.7 cloned from the genomic DNA of Conus textile that inhibits calcium currents Maojun Zhou Manyi Yang Huiling Wen Shun Xu Cuifang Han Yun Wu 2023-05-01T00:00:00Z https://doi.org/10.1590/1678-9199-jvatitd-2022-0085 https://doaj.org/article/005360dd729644999e601e2008bca650 EN eng SciELO http://www.scielo.br/scielo.php?script=sci_arttext&pid=S1678-91992023000100306&lng=en&tlng=en http://www.scielo.br/pdf/jvatitd/v29/1678-9199-jvatitd-28-e20220085.pdf https://doaj.org/toc/1678-9199 1678-9199 doi:10.1590/1678-9199-jvatitd-2022-0085 https://doaj.org/article/005360dd729644999e601e2008bca650 Journal of Venomous Animals and Toxins including Tropical Diseases, Vol 29 (2023) conotoxin Tx6.7 calcium currents hCaV1.2 hCaV2.2 Conus textile Arctic medicine. Tropical medicine RC955-962 Toxicology. Poisons RA1190-1270 Zoology QL1-991 article 2023 ftdoajarticles https://doi.org/10.1590/1678-9199-jvatitd-2022-0085 2023-05-28T00:37:06Z Abstract Background: Conotoxins exhibit great potential as neuropharmacology tools and therapeutic candidates due to their high affinity and specificity for ion channels, neurotransmitter receptors or transporters. The traditional methods to discover new conotoxins are peptide purification from the crude venom or gene amplification from the venom duct. Methods: In this study, a novel O1 superfamily conotoxin Tx6.7 was directly cloned from the genomic DNA of Conus textile using primers corresponding to the conserved intronic sequence and 3’ UTR elements. The mature peptide of Tx6.7 (DCHERWDWCPASLLGVIYCCEGLICFIAFCI) was synthesized by solid-phase chemical synthesis and confirmed by mass spectrometry. Results: Patch clamp experiments on rat DRG neurons showed that Tx6.7 inhibited peak calcium currents by 59.29 ± 2.34% and peak potassium currents by 22.33 ± 7.81%. In addition, patch clamp on the ion channel subtypes showed that 10 μM Tx6.7 inhibited 56.61 ± 3.20% of the hCaV1.2 currents, 24.67 ± 0.91% of the hCaV2.2 currents and 7.30 ± 3.38% of the hNaV1.8 currents. Tx6.7 had no significant toxicity to ND7/23 cells and increased the pain threshold from 0.5 to 4 hours in the mouse hot plate assay. Conclusion: Our results suggested that direct cloning of conotoxin sequences from the genomic DNA of cone snails would be an alternative approach to obtaining novel conotoxins. Tx6.7 could be used as a probe tool for ion channel research or a therapeutic candidate for novel drug development. Article in Journal/Newspaper Arctic Directory of Open Access Journals: DOAJ Articles Arctic Journal of Venomous Animals and Toxins including Tropical Diseases 29 |
institution |
Open Polar |
collection |
Directory of Open Access Journals: DOAJ Articles |
op_collection_id |
ftdoajarticles |
language |
English |
topic |
conotoxin Tx6.7 calcium currents hCaV1.2 hCaV2.2 Conus textile Arctic medicine. Tropical medicine RC955-962 Toxicology. Poisons RA1190-1270 Zoology QL1-991 |
spellingShingle |
conotoxin Tx6.7 calcium currents hCaV1.2 hCaV2.2 Conus textile Arctic medicine. Tropical medicine RC955-962 Toxicology. Poisons RA1190-1270 Zoology QL1-991 Maojun Zhou Manyi Yang Huiling Wen Shun Xu Cuifang Han Yun Wu O1-conotoxin Tx6.7 cloned from the genomic DNA of Conus textile that inhibits calcium currents |
topic_facet |
conotoxin Tx6.7 calcium currents hCaV1.2 hCaV2.2 Conus textile Arctic medicine. Tropical medicine RC955-962 Toxicology. Poisons RA1190-1270 Zoology QL1-991 |
description |
Abstract Background: Conotoxins exhibit great potential as neuropharmacology tools and therapeutic candidates due to their high affinity and specificity for ion channels, neurotransmitter receptors or transporters. The traditional methods to discover new conotoxins are peptide purification from the crude venom or gene amplification from the venom duct. Methods: In this study, a novel O1 superfamily conotoxin Tx6.7 was directly cloned from the genomic DNA of Conus textile using primers corresponding to the conserved intronic sequence and 3’ UTR elements. The mature peptide of Tx6.7 (DCHERWDWCPASLLGVIYCCEGLICFIAFCI) was synthesized by solid-phase chemical synthesis and confirmed by mass spectrometry. Results: Patch clamp experiments on rat DRG neurons showed that Tx6.7 inhibited peak calcium currents by 59.29 ± 2.34% and peak potassium currents by 22.33 ± 7.81%. In addition, patch clamp on the ion channel subtypes showed that 10 μM Tx6.7 inhibited 56.61 ± 3.20% of the hCaV1.2 currents, 24.67 ± 0.91% of the hCaV2.2 currents and 7.30 ± 3.38% of the hNaV1.8 currents. Tx6.7 had no significant toxicity to ND7/23 cells and increased the pain threshold from 0.5 to 4 hours in the mouse hot plate assay. Conclusion: Our results suggested that direct cloning of conotoxin sequences from the genomic DNA of cone snails would be an alternative approach to obtaining novel conotoxins. Tx6.7 could be used as a probe tool for ion channel research or a therapeutic candidate for novel drug development. |
format |
Article in Journal/Newspaper |
author |
Maojun Zhou Manyi Yang Huiling Wen Shun Xu Cuifang Han Yun Wu |
author_facet |
Maojun Zhou Manyi Yang Huiling Wen Shun Xu Cuifang Han Yun Wu |
author_sort |
Maojun Zhou |
title |
O1-conotoxin Tx6.7 cloned from the genomic DNA of Conus textile that inhibits calcium currents |
title_short |
O1-conotoxin Tx6.7 cloned from the genomic DNA of Conus textile that inhibits calcium currents |
title_full |
O1-conotoxin Tx6.7 cloned from the genomic DNA of Conus textile that inhibits calcium currents |
title_fullStr |
O1-conotoxin Tx6.7 cloned from the genomic DNA of Conus textile that inhibits calcium currents |
title_full_unstemmed |
O1-conotoxin Tx6.7 cloned from the genomic DNA of Conus textile that inhibits calcium currents |
title_sort |
o1-conotoxin tx6.7 cloned from the genomic dna of conus textile that inhibits calcium currents |
publisher |
SciELO |
publishDate |
2023 |
url |
https://doi.org/10.1590/1678-9199-jvatitd-2022-0085 https://doaj.org/article/005360dd729644999e601e2008bca650 |
geographic |
Arctic |
geographic_facet |
Arctic |
genre |
Arctic |
genre_facet |
Arctic |
op_source |
Journal of Venomous Animals and Toxins including Tropical Diseases, Vol 29 (2023) |
op_relation |
http://www.scielo.br/scielo.php?script=sci_arttext&pid=S1678-91992023000100306&lng=en&tlng=en http://www.scielo.br/pdf/jvatitd/v29/1678-9199-jvatitd-28-e20220085.pdf https://doaj.org/toc/1678-9199 1678-9199 doi:10.1590/1678-9199-jvatitd-2022-0085 https://doaj.org/article/005360dd729644999e601e2008bca650 |
op_doi |
https://doi.org/10.1590/1678-9199-jvatitd-2022-0085 |
container_title |
Journal of Venomous Animals and Toxins including Tropical Diseases |
container_volume |
29 |
_version_ |
1768383618885877760 |