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...
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| Format: | Article |
| Language: | English |
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SciELO
2020-05-01
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| Series: | Journal of Venomous Animals and Toxins including Tropical Diseases |
| Subjects: | |
| Online Access: | http://www.scielo.br/scielo.php?script=sci_arttext&pid=S1678-91992020000100311&tlng=en |
| _version_ | 1798021380989517824 |
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| 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 |
| collection | DOAJ |
| 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 require further investigation. |
| first_indexed | 2024-04-11T17:13:00Z |
| format | Article |
| id | doaj.art-bac14acb66ae4287a75db1ee5c7d5e72 |
| institution | Directory Open Access Journal |
| issn | 1678-9199 |
| language | English |
| last_indexed | 2024-04-11T17:13:00Z |
| publishDate | 2020-05-01 |
| publisher | SciELO |
| record_format | Article |
| series | Journal of Venomous Animals and Toxins including Tropical Diseases |
| spelling | doaj.art-bac14acb66ae4287a75db1ee5c7d5e722022-12-22T04:12:51ZengSciELOJournal of Venomous Animals and Toxins including Tropical Diseases1678-91992020-05-012610.1590/1678-9199-jvatitd-2019-0095Isolation and structural identification of a new T1-conotoxin with unique disulfide connectivities derived from Conus bandanusNguyen Baohttps://orcid.org/0000-0001-7122-8510Jean-Pière LecaerNgo Dang NghiaPhan Thi Khanh VinhAbstract 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 require further investigation.http://www.scielo.br/scielo.php?script=sci_arttext&pid=S1678-91992020000100311&tlng=enT1-subfamily conotoxinConus bandanusBn5aDisulfide connectivityCone snail venom |
| spellingShingle | 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 Journal of Venomous Animals and Toxins including Tropical Diseases T1-subfamily conotoxin Conus bandanus Bn5a Disulfide connectivity Cone snail venom |
| title | 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_short | 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 |
| topic | T1-subfamily conotoxin Conus bandanus Bn5a Disulfide connectivity Cone snail venom |
| url | http://www.scielo.br/scielo.php?script=sci_arttext&pid=S1678-91992020000100311&tlng=en |
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