Isolation and structural identification of a new T1-conotoxin with unique disulfide connectivities derived from Conus bandanus
Abstract Background: Conopeptides are neuropharmacological peptides derived from the venomous salivary glands of cone snails. Among 29 superfamilies based on conserved signal sequences, T-superfamily conotoxins, which belong to the smallest group, include four different frameworks that contain four...
Published in: | Journal of Venomous Animals and Toxins including Tropical Diseases |
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ftdoajarticles:oai:doaj.org/article:bac14acb66ae4287a75db1ee5c7d5e72 2023-05-15T15:15:51+02:00 Isolation and structural identification of a new T1-conotoxin with unique disulfide connectivities derived from Conus bandanus Nguyen Bao Jean-Pière Lecaer Ngo Dang Nghia Phan Thi Khanh Vinh 2020-05-01T00:00:00Z https://doi.org/10.1590/1678-9199-jvatitd-2019-0095 https://doaj.org/article/bac14acb66ae4287a75db1ee5c7d5e72 EN eng SciELO http://www.scielo.br/scielo.php?script=sci_arttext&pid=S1678-91992020000100311&tlng=en http://www.scielo.br/pdf/jvatitd/v26/1678-9199-jvatitd-26-e20190095.pdf https://doaj.org/toc/1678-9199 1678-9199 doi:10.1590/1678-9199-jvatitd-2019-0095 https://doaj.org/article/bac14acb66ae4287a75db1ee5c7d5e72 Journal of Venomous Animals and Toxins including Tropical Diseases, Vol 26 (2020) T1-subfamily conotoxin Conus bandanus Bn5a Disulfide connectivity Cone snail venom Arctic medicine. Tropical medicine RC955-962 Toxicology. Poisons RA1190-1270 Zoology QL1-991 article 2020 ftdoajarticles https://doi.org/10.1590/1678-9199-jvatitd-2019-0095 2022-12-30T20:22:20Z Abstract Background: Conopeptides are neuropharmacological peptides derived from the venomous salivary glands of cone snails. Among 29 superfamilies based on conserved signal sequences, T-superfamily conotoxins, which belong to the smallest group, include four different frameworks that contain four cysteines denominated I, V, X and XVI. In this work, the primary structure and the cysteine connectivity of novel conotoxin of Conus bandanus were determined by tandem mass spectrometry using collision-induced dissociation. Methods: The venom glands of C. bandanus snails were dissected, pooled, and extracted with 0.1% trifluoroacetic acid in three steps and lyophilized. The venom was fractionated and purified in an HPLC system with an analytical reversed-phase C18 column. The primary peptide structure was analyzed by MALDI TOF MS/MS using collision-induced dissociation and confirmed by Edman's degradation. The peptide’s cysteine connectivity was determined by rapid partial reduction-alkylation technique. Results: The novel conotoxin, NGC1C2(I/L)VREC3C4, was firstly derived from de novo sequencing by MS/MS. The presence of isoleucine residues in this conotoxin was confirmed by the Edman degradation method. The conotoxin, denominated Bn5a, belongs to the T1-subfamily of conotoxins. However, the disulfide bonds (C1-C4/C2-C3) of Bn5a were not the same as found in other T1-subfamily conopeptides but shared common connectivities with T2-subfamily conotoxins. The T1-conotoxin of C. bandanus proved the complexity of the disulfide bond pattern of conopeptides. The homological analysis revealed that the novel conotoxin could serve as a valuable probe compound for the human-nervous-system norepinephrine transporter. Conclusion: We identified the first T1-conotoxin, denominated Bn5a, isolated from C. bandanus venom. However, Bn5a conotoxin exhibited unique C1-C4/C2-C3 disulfide connectivity, unlike other T1-conotoxins (C1-C3/C2-C4). The structural and homological analyses herein have evidenced novel conotoxin Bn5a that may ... Article in Journal/Newspaper Arctic Directory of Open Access Journals: DOAJ Articles Arctic Journal of Venomous Animals and Toxins including Tropical Diseases 26 |
institution |
Open Polar |
collection |
Directory of Open Access Journals: DOAJ Articles |
op_collection_id |
ftdoajarticles |
language |
English |
topic |
T1-subfamily conotoxin Conus bandanus Bn5a Disulfide connectivity Cone snail venom Arctic medicine. Tropical medicine RC955-962 Toxicology. Poisons RA1190-1270 Zoology QL1-991 |
spellingShingle |
T1-subfamily conotoxin Conus bandanus Bn5a Disulfide connectivity Cone snail venom Arctic medicine. Tropical medicine RC955-962 Toxicology. Poisons RA1190-1270 Zoology QL1-991 Nguyen Bao Jean-Pière Lecaer Ngo Dang Nghia Phan Thi Khanh Vinh Isolation and structural identification of a new T1-conotoxin with unique disulfide connectivities derived from Conus bandanus |
topic_facet |
T1-subfamily conotoxin Conus bandanus Bn5a Disulfide connectivity Cone snail venom Arctic medicine. Tropical medicine RC955-962 Toxicology. Poisons RA1190-1270 Zoology QL1-991 |
description |
Abstract Background: Conopeptides are neuropharmacological peptides derived from the venomous salivary glands of cone snails. Among 29 superfamilies based on conserved signal sequences, T-superfamily conotoxins, which belong to the smallest group, include four different frameworks that contain four cysteines denominated I, V, X and XVI. In this work, the primary structure and the cysteine connectivity of novel conotoxin of Conus bandanus were determined by tandem mass spectrometry using collision-induced dissociation. Methods: The venom glands of C. bandanus snails were dissected, pooled, and extracted with 0.1% trifluoroacetic acid in three steps and lyophilized. The venom was fractionated and purified in an HPLC system with an analytical reversed-phase C18 column. The primary peptide structure was analyzed by MALDI TOF MS/MS using collision-induced dissociation and confirmed by Edman's degradation. The peptide’s cysteine connectivity was determined by rapid partial reduction-alkylation technique. Results: The novel conotoxin, NGC1C2(I/L)VREC3C4, was firstly derived from de novo sequencing by MS/MS. The presence of isoleucine residues in this conotoxin was confirmed by the Edman degradation method. The conotoxin, denominated Bn5a, belongs to the T1-subfamily of conotoxins. However, the disulfide bonds (C1-C4/C2-C3) of Bn5a were not the same as found in other T1-subfamily conopeptides but shared common connectivities with T2-subfamily conotoxins. The T1-conotoxin of C. bandanus proved the complexity of the disulfide bond pattern of conopeptides. The homological analysis revealed that the novel conotoxin could serve as a valuable probe compound for the human-nervous-system norepinephrine transporter. Conclusion: We identified the first T1-conotoxin, denominated Bn5a, isolated from C. bandanus venom. However, Bn5a conotoxin exhibited unique C1-C4/C2-C3 disulfide connectivity, unlike other T1-conotoxins (C1-C3/C2-C4). The structural and homological analyses herein have evidenced novel conotoxin Bn5a that may ... |
format |
Article in Journal/Newspaper |
author |
Nguyen Bao Jean-Pière Lecaer Ngo Dang Nghia Phan Thi Khanh Vinh |
author_facet |
Nguyen Bao Jean-Pière Lecaer Ngo Dang Nghia Phan Thi Khanh Vinh |
author_sort |
Nguyen Bao |
title |
Isolation and structural identification of a new T1-conotoxin with unique disulfide connectivities derived from Conus bandanus |
title_short |
Isolation and structural identification of a new T1-conotoxin with unique disulfide connectivities derived from Conus bandanus |
title_full |
Isolation and structural identification of a new T1-conotoxin with unique disulfide connectivities derived from Conus bandanus |
title_fullStr |
Isolation and structural identification of a new T1-conotoxin with unique disulfide connectivities derived from Conus bandanus |
title_full_unstemmed |
Isolation and structural identification of a new T1-conotoxin with unique disulfide connectivities derived from Conus bandanus |
title_sort |
isolation and structural identification of a new t1-conotoxin with unique disulfide connectivities derived from conus bandanus |
publisher |
SciELO |
publishDate |
2020 |
url |
https://doi.org/10.1590/1678-9199-jvatitd-2019-0095 https://doaj.org/article/bac14acb66ae4287a75db1ee5c7d5e72 |
geographic |
Arctic |
geographic_facet |
Arctic |
genre |
Arctic |
genre_facet |
Arctic |
op_source |
Journal of Venomous Animals and Toxins including Tropical Diseases, Vol 26 (2020) |
op_relation |
http://www.scielo.br/scielo.php?script=sci_arttext&pid=S1678-91992020000100311&tlng=en http://www.scielo.br/pdf/jvatitd/v26/1678-9199-jvatitd-26-e20190095.pdf https://doaj.org/toc/1678-9199 1678-9199 doi:10.1590/1678-9199-jvatitd-2019-0095 https://doaj.org/article/bac14acb66ae4287a75db1ee5c7d5e72 |
op_doi |
https://doi.org/10.1590/1678-9199-jvatitd-2019-0095 |
container_title |
Journal of Venomous Animals and Toxins including Tropical Diseases |
container_volume |
26 |
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1766346188088410112 |