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Proteome Based de novo Sequencing of Novel Conotoxins from Marine Molluscivorous Cone Snail Conus amadis and Neurological Activities of Its Natural Venom in Zebrafish Model

Rajesh, RP and Franklin, Jayaseelan B and Badsha, Iffath and Arjun, P and Jain, Ruchi P and Vignesh, M S and Kannan, Rajesh R (2019) Proteome Based de novo Sequencing of Novel Conotoxins from Marine Molluscivorous Cone Snail Conus amadis and Neurological Activities of Its Natural Venom in Zebrafish Model. In: PROTEIN AND PEPTIDE LETTERS, 26 (11). pp. 819-833.

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Official URL: https://dx.doi.org/10.2174/09298665266661906141440...


Background: Conus amadis is a carnivorous snail found abundantly in coastal waters of India. Despite its abundance in southern coastal waters of India and the fact that most of the conotoxin act in neuronal system, research work on Conus amadis venom was not much focused. So we have made a brief study on the venom complex of Conus amadis to identify the library of novel conotoxins and to screen the natural venom for neurological function. Objective: De novo sequencing of novel conopeptides from the venom cocktail of Conus amadis and to screen its natural venom for the presence of biological activities in zebrafish model. Methods: Proteome based MALDI-TOF and LC-MS-MS analysis for identification of novel conotoxins and subsequent sequencing. Due to the complex disulfide rich nature of the venom peptides, the study also involves global chemical modification experiments of the venom extract to unambiguously determine the sequence of novel conotoxins. Biological function analysis of natural venom was tested in zebrafish model to ascertain anti-epileptic properties. Results: In this study, we have identified 19 novel conotox ins containing 1, 2 & 3 disulfides, belonging to different classes. Among them, 2 novel contryphans, 3 T-superfamily conotoxins, 2 A-superfamily conotoxins and 2 Mini M-Superfamily conotoxins were sequenced to its amino acid level from the fragmented spectrum of singly and doubly charged parent ions using de novo sequencing strategies. ama1054, a contryphan peptide toxin, possesses post translationally modified bromo tryptophan at its seventh position. Except ama1251, all the sequenced peptide toxins possess modified C-terminal amidation. Crude venom exhibited anticonvulsant properties in pentylenetetrazole-induced seizure in zebrafish larvae, which suggested anti-epileptic property of the venom cocktail. Acetylcholinesterase activity was also identified in the venom complex. Conclusion: Based on the preliminary evidence, if this study is extended further through bioassay guided purification, could possibly yield peptide toxins with anticonvulsant and other neurologically active molecules.

Item Type: Journal Article
Additional Information: copyright for this article belongs to BENTHAM SCIENCE PUBL LTD
Keywords: Conus amadis; conotoxin; antiepileptic drug; mass spectrometry; zebrafish; acetylcholinesterase activity
Department/Centre: Division of Biological Sciences > Molecular Biophysics Unit
Date Deposited: 15 Nov 2019 05:02
Last Modified: 17 Nov 2019 15:57
URI: http://eprints.iisc.ac.in/id/eprint/63908

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