Isolation and structural identification of a new T1-conotoxin with unique disulfide connectivities derived from Conus bandanus
| Autor: | Ngo Dang Nghia, Phan Thi Khanh Vinh, Jean-Pière Lecaer, Nguyen Bao |
|---|---|
| Jazyk: | angličtina |
| Rok vydání: | 2020 |
| Předmět: |
0301 basic medicine
Conus bandanus Stereochemistry Disulfide connectivity 030231 tropical medicine RC955-962 Venom Peptide Cone snail venom Toxicology Tandem mass spectrometry complex mixtures T1-subfamily conotoxin 03 medical and health sciences 0302 clinical medicine Arctic medicine. Tropical medicine RA1190-1270 Conotoxin chemistry.chemical_classification 030102 biochemistry & molecular biology Edman degradation biology Research Protein primary structure biology.organism_classification Bn5a Infectious Diseases chemistry QL1-991 Toxicology. Poisons Animal Science and Zoology Parasitology Zoology Cysteine |
| Zdroj: | Journal of Venomous Animals and Toxins including Tropical Diseases, Volume: 26, Article number: e20190095, Published: 08 MAY 2020 Journal of Venomous Animals and Toxins including Tropical Diseases, Vol 26 (2020) Journal of Venomous Animals and Toxins including Tropical Diseases v.26 2020 The Journal of venomous animals and toxins including tropical diseases Universidade Estadual Paulista (UNESP) instacron:UNESP The Journal of Venomous Animals and Toxins Including Tropical Diseases |
| Popis: | 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. |
| Databáze: | OpenAIRE |
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