Unveiling the functional epitopes of cobra venom cytotoxin by immunoinformatics and epitope-omic analyses
Abstract Approximate 70% of cobra venom is composed of cytotoxin (CTX), which is responsible for the dermonecrotic symptoms of cobra envenomation. However, CTX is generally low in immunogenicity, and the antivenom is ineffective in attenuating its in vivo toxicity. Furthermore, little is known about...
| Main Authors: | , , , |
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| Format: | Article |
| Language: | English |
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Nature Portfolio
2023-07-01
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| Series: | Scientific Reports |
| Online Access: | https://doi.org/10.1038/s41598-023-39222-2 |
| _version_ | 1827890375481622528 |
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| author | Jia Jin Hiu Jared Kah Yin Fung Hock Siew Tan Michelle Khai Khun Yap |
| author_facet | Jia Jin Hiu Jared Kah Yin Fung Hock Siew Tan Michelle Khai Khun Yap |
| author_sort | Jia Jin Hiu |
| collection | DOAJ |
| description | Abstract Approximate 70% of cobra venom is composed of cytotoxin (CTX), which is responsible for the dermonecrotic symptoms of cobra envenomation. However, CTX is generally low in immunogenicity, and the antivenom is ineffective in attenuating its in vivo toxicity. Furthermore, little is known about its epitope properties for empirical antivenom therapy. This study aimed to determine the epitope sequences of CTX using the immunoinformatic analyses and epitope-omics profiling. A conserved CTX was used in this study to determine its T-cell and B-cell epitope sequences using immunoinformatic tools and molecular docking simulation with different Human Leukocyte Antigens (HLAs). The potential T-cell and B-cell epitopes were 'KLVPLFY,' 'CPAGKNLCY,' 'MFMVSTPTK,' and 'DVCPKNSLL.' Molecular docking simulations disclosed that the HLA-B62 supertype exhibited the greatest binding affinity towards cobra venom cytotoxin. The namely L7, G18, K19, N20, M25, K33, V43, C44, K46, N47, and S48 of CTX exhibited prominent intermolecular interactions with HLA-B62. The multi-enzymatic-limited-digestion/liquid chromatography-mass spectrometry (MELD/LC–MS) also revealed three potential epitope sequences as 'LVPLFYK,' 'MFMVS,' and ‘TVPVKR’. From different epitope mapping approaches, we concluded four potential epitope sites of CTX as ‘KLVPLFYK’, ‘AGKNL’, ‘MFMVSTPKVPV’ and ‘DVCPKNSLL’. Site-directed mutagenesis of these epitopes confirmed their locations at the functional loops of CTX. These epitope sequences are crucial to CTX’s structural folding and cytotoxicity. The results concluded the epitopes that resided within the functional loops constituted potential targets to fabricate synthetic epitopes for CTX-targeted antivenom production. |
| first_indexed | 2024-03-12T21:09:52Z |
| format | Article |
| id | doaj.art-3e9d1e8f62b5427b8a5ba267bb0310c5 |
| institution | Directory Open Access Journal |
| issn | 2045-2322 |
| language | English |
| last_indexed | 2024-03-12T21:09:52Z |
| publishDate | 2023-07-01 |
| publisher | Nature Portfolio |
| record_format | Article |
| series | Scientific Reports |
| spelling | doaj.art-3e9d1e8f62b5427b8a5ba267bb0310c52023-07-30T11:15:41ZengNature PortfolioScientific Reports2045-23222023-07-0113111410.1038/s41598-023-39222-2Unveiling the functional epitopes of cobra venom cytotoxin by immunoinformatics and epitope-omic analysesJia Jin Hiu0Jared Kah Yin Fung1Hock Siew Tan2Michelle Khai Khun Yap3School of Science, Monash University MalaysiaSchool of Science, Monash University MalaysiaSchool of Science, Monash University MalaysiaSchool of Science, Monash University MalaysiaAbstract Approximate 70% of cobra venom is composed of cytotoxin (CTX), which is responsible for the dermonecrotic symptoms of cobra envenomation. However, CTX is generally low in immunogenicity, and the antivenom is ineffective in attenuating its in vivo toxicity. Furthermore, little is known about its epitope properties for empirical antivenom therapy. This study aimed to determine the epitope sequences of CTX using the immunoinformatic analyses and epitope-omics profiling. A conserved CTX was used in this study to determine its T-cell and B-cell epitope sequences using immunoinformatic tools and molecular docking simulation with different Human Leukocyte Antigens (HLAs). The potential T-cell and B-cell epitopes were 'KLVPLFY,' 'CPAGKNLCY,' 'MFMVSTPTK,' and 'DVCPKNSLL.' Molecular docking simulations disclosed that the HLA-B62 supertype exhibited the greatest binding affinity towards cobra venom cytotoxin. The namely L7, G18, K19, N20, M25, K33, V43, C44, K46, N47, and S48 of CTX exhibited prominent intermolecular interactions with HLA-B62. The multi-enzymatic-limited-digestion/liquid chromatography-mass spectrometry (MELD/LC–MS) also revealed three potential epitope sequences as 'LVPLFYK,' 'MFMVS,' and ‘TVPVKR’. From different epitope mapping approaches, we concluded four potential epitope sites of CTX as ‘KLVPLFYK’, ‘AGKNL’, ‘MFMVSTPKVPV’ and ‘DVCPKNSLL’. Site-directed mutagenesis of these epitopes confirmed their locations at the functional loops of CTX. These epitope sequences are crucial to CTX’s structural folding and cytotoxicity. The results concluded the epitopes that resided within the functional loops constituted potential targets to fabricate synthetic epitopes for CTX-targeted antivenom production.https://doi.org/10.1038/s41598-023-39222-2 |
| spellingShingle | Jia Jin Hiu Jared Kah Yin Fung Hock Siew Tan Michelle Khai Khun Yap Unveiling the functional epitopes of cobra venom cytotoxin by immunoinformatics and epitope-omic analyses Scientific Reports |
| title | Unveiling the functional epitopes of cobra venom cytotoxin by immunoinformatics and epitope-omic analyses |
| title_full | Unveiling the functional epitopes of cobra venom cytotoxin by immunoinformatics and epitope-omic analyses |
| title_fullStr | Unveiling the functional epitopes of cobra venom cytotoxin by immunoinformatics and epitope-omic analyses |
| title_full_unstemmed | Unveiling the functional epitopes of cobra venom cytotoxin by immunoinformatics and epitope-omic analyses |
| title_short | Unveiling the functional epitopes of cobra venom cytotoxin by immunoinformatics and epitope-omic analyses |
| title_sort | unveiling the functional epitopes of cobra venom cytotoxin by immunoinformatics and epitope omic analyses |
| url | https://doi.org/10.1038/s41598-023-39222-2 |
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