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Identification of Potential Insulinotropic Cytotoxins from Indian Cobra Snake Venom Using High-Resolution Mass Spectrometry and Analyzing Their Possible Interactions with Potassium Channel Receptors by In Silico Studies

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Abstract

Snake venoms are a potential source of bioactive peptides, which have multiple therapeutic properties in treating diseases such as diabetes, cancer, and neurological disorders. Among bioactive peptides, cytotoxins (CTXs) and neurotoxins are low molecular weight proteins belonging to the three-finger-fold toxins (3FTxs) family composed of two β sheets that are stabilized by four to five conserved disulfide bonds containing 58–72 amino acid residues. These are highly abundant in snake venom and are predicted to have insulinotropic activities. In this study, the CTXs were purified from Indian cobra snake venom using preparative HPLC and characterized using high-resolution mass spectrometry (HRMS) TOF–MS/MS. Further SDS-PAGE analysis confirmed the presence of low molecular weight cytotoxic proteins. The CTXs in fractions A and B exhibited dose-dependent insulinotropic activity from 0.001 to 10 µM using rat pancreatic beta-cell lines (RIN-5F) in the ELISA. Nateglinide and repaglinide are synthetic small-molecule drugs that control sugar levels in the blood in type 2 diabetes, which were used as a positive control in ELISA. Concluded that purified CTXs have insulinotropic activity, and there is a scope to use these proteins as small molecules to stimulate insulinotropic activities. At this stage, the focus is on the efficiency of the cytotoxins to induce insulin. Additional work is ongoing on animal models to see the extent of the beneficial effects and efficiency to cure diabetes using streptozotocin-induced models.

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Data Availability

All datasets generated for this study are included in the article/Supplementary Material.

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Acknowledgements

This work is partly supported by the Kyntox Biotech India Pvt Ltd, Bangalore, Bangalore Bio-innovation Center, SCIEX, India and BioNome, Bangalore.

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Authors and Affiliations

  1. Department of Biotechnology, Dravidian University, Kuppam, 517426, Andhra Pradesh, India

    Upendra Gunta & M. Mastan

  2. Kyntox Biotech India Pvt Ltd, Bangalore, 560032, Karnakata, India

    Gangadhar P. Vadla & Gopi Kadiyala

  3. Shri Jagdishprasad Jhabarmal Tibrewala University, Jhunjhunu, 333010, Rajasthan, India

    Dilipkumar Reddy Kandula

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  1. Upendra Gunta
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Contributions

Upendra Gunta: Data curation; Formal analysis; Conceptualization; Investigation; Validation; Writing – original draft, reviewing and editing the manuscript. Gangadhar P Vadla: Formal analysis; Conceptualization; Investigation; Supervision, review and editing. Gopi Kadiyala: Formal analysis; Review. Dilipkumar Reddy Kandula: Formal analysis; Review and editing. M. Mastan: Methodology; Conceptualization; Investigation; Supervision, Validation; Writing–original draft; Writing–review and editing.

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Correspondence to M. Mastan.

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Gunta, U., Vadla, G.P., Kadiyala, G. et al. Identification of Potential Insulinotropic Cytotoxins from Indian Cobra Snake Venom Using High-Resolution Mass Spectrometry and Analyzing Their Possible Interactions with Potassium Channel Receptors by In Silico Studies. Appl Biochem Biotechnol 196, 160–181 (2024). https://doi.org/10.1007/s12010-023-04523-9

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