Heterologous Expression and Immunogenic Potential of the Most Abundant Phospholipase A<sub>2</sub> from Coral Snake <i>Micrurus dumerilii</i> to Develop Antivenoms
<i>Micrurus dumerilii</i> is a coral snake of clinic interest in Colombia. Its venom is mainly composed of phospholipases A<sub>2</sub> being MdumPLA<sub>2</sub> the most abundant protein. Nevertheless, <i>Micrurus</i> species produce a low quantity of...
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MDPI AG
2022-11-01
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| Series: | Toxins |
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| Online Access: | https://www.mdpi.com/2072-6651/14/12/825 |
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| author | Luz E. Romero-Giraldo Sergio Pulido Mario A. Berrío María F. Flórez Paola Rey-Suárez Vitelbina Nuñez Jaime A. Pereañez |
| author_facet | Luz E. Romero-Giraldo Sergio Pulido Mario A. Berrío María F. Flórez Paola Rey-Suárez Vitelbina Nuñez Jaime A. Pereañez |
| author_sort | Luz E. Romero-Giraldo |
| collection | DOAJ |
| description | <i>Micrurus dumerilii</i> is a coral snake of clinic interest in Colombia. Its venom is mainly composed of phospholipases A<sub>2</sub> being MdumPLA<sub>2</sub> the most abundant protein. Nevertheless, <i>Micrurus</i> species produce a low quantity of venom, which makes it difficult to produce anticoral antivenoms. Therefore, in this work, we present the recombinant expression of MdumPLA<sub>2</sub> to evaluate its biological activities and its immunogenic potential to produce antivenoms. For this, a genetic construct rMdumPLA<sub>2</sub> was cloned into the pET28a vector and expressed heterologously in bacteria. His-rMdumPLA<sub>2</sub> was extracted from inclusion bodies, refolded in vitro, and isolated using affinity and RP-HPLC chromatography. His-rMdumPLA<sub>2</sub> was shown to have phospholipase A<sub>2</sub> activity, a weak anticoagulant effect, and induced myonecrosis and edema. The anti-His-rMdumPLA<sub>2</sub> antibodies produced in rabbits recognized native PLA<sub>2</sub>, the complete venom of <i>M. dumerilii</i>, and a phospholipase from another species of the <i>Micrurus</i> genus. Antibodies neutralized 100% of the in vitro phospholipase activity of the recombinant toxin and a moderate percentage of the myotoxic activity of <i>M. dumerilii</i> venom in mice. These results indicate that His-rMdumPLA<sub>2</sub> could be used as an immunogen to improve anticoral antivenoms development. This work is the first report of an <i>M. dumerilii</i> functional recombinant PLA<sub>2</sub>. |
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| institution | Directory Open Access Journal |
| issn | 2072-6651 |
| language | English |
| last_indexed | 2024-03-09T15:46:24Z |
| publishDate | 2022-11-01 |
| publisher | MDPI AG |
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| series | Toxins |
| spelling | doaj.art-1686a3e0b2074197bd5110cfce0eef5a2023-11-24T18:26:25ZengMDPI AGToxins2072-66512022-11-01141282510.3390/toxins14120825Heterologous Expression and Immunogenic Potential of the Most Abundant Phospholipase A<sub>2</sub> from Coral Snake <i>Micrurus dumerilii</i> to Develop AntivenomsLuz E. Romero-Giraldo0Sergio Pulido1Mario A. Berrío2María F. Flórez3Paola Rey-Suárez4Vitelbina Nuñez5Jaime A. Pereañez6Research Group in Toxinology, Pharmaceutical, and Food Alternatives, Pharmaceutical and Food Sciences Faculty, University of Antioquia, Medellín 50010, ColombiaTropical Disease Study and Control Program—PECET, University of Antioquia, Medellín 50010, ColombiaTropical Disease Study and Control Program—PECET, University of Antioquia, Medellín 50010, ColombiaTropical Disease Study and Control Program—PECET, University of Antioquia, Medellín 50010, ColombiaResearch Group in Toxinology, Pharmaceutical, and Food Alternatives, Pharmaceutical and Food Sciences Faculty, University of Antioquia, Medellín 50010, ColombiaResearch Group in Toxinology, Pharmaceutical, and Food Alternatives, Pharmaceutical and Food Sciences Faculty, University of Antioquia, Medellín 50010, ColombiaResearch Group in Toxinology, Pharmaceutical, and Food Alternatives, Pharmaceutical and Food Sciences Faculty, University of Antioquia, Medellín 50010, Colombia<i>Micrurus dumerilii</i> is a coral snake of clinic interest in Colombia. Its venom is mainly composed of phospholipases A<sub>2</sub> being MdumPLA<sub>2</sub> the most abundant protein. Nevertheless, <i>Micrurus</i> species produce a low quantity of venom, which makes it difficult to produce anticoral antivenoms. Therefore, in this work, we present the recombinant expression of MdumPLA<sub>2</sub> to evaluate its biological activities and its immunogenic potential to produce antivenoms. For this, a genetic construct rMdumPLA<sub>2</sub> was cloned into the pET28a vector and expressed heterologously in bacteria. His-rMdumPLA<sub>2</sub> was extracted from inclusion bodies, refolded in vitro, and isolated using affinity and RP-HPLC chromatography. His-rMdumPLA<sub>2</sub> was shown to have phospholipase A<sub>2</sub> activity, a weak anticoagulant effect, and induced myonecrosis and edema. The anti-His-rMdumPLA<sub>2</sub> antibodies produced in rabbits recognized native PLA<sub>2</sub>, the complete venom of <i>M. dumerilii</i>, and a phospholipase from another species of the <i>Micrurus</i> genus. Antibodies neutralized 100% of the in vitro phospholipase activity of the recombinant toxin and a moderate percentage of the myotoxic activity of <i>M. dumerilii</i> venom in mice. These results indicate that His-rMdumPLA<sub>2</sub> could be used as an immunogen to improve anticoral antivenoms development. This work is the first report of an <i>M. dumerilii</i> functional recombinant PLA<sub>2</sub>.https://www.mdpi.com/2072-6651/14/12/825recombinant proteinantibodies<i>Micrurus dumerilii</i>coral snake antivenomsphospholipase A<sub>2</sub> |
| spellingShingle | Luz E. Romero-Giraldo Sergio Pulido Mario A. Berrío María F. Flórez Paola Rey-Suárez Vitelbina Nuñez Jaime A. Pereañez Heterologous Expression and Immunogenic Potential of the Most Abundant Phospholipase A<sub>2</sub> from Coral Snake <i>Micrurus dumerilii</i> to Develop Antivenoms Toxins recombinant protein antibodies <i>Micrurus dumerilii</i> coral snake antivenoms phospholipase A<sub>2</sub> |
| title | Heterologous Expression and Immunogenic Potential of the Most Abundant Phospholipase A<sub>2</sub> from Coral Snake <i>Micrurus dumerilii</i> to Develop Antivenoms |
| title_full | Heterologous Expression and Immunogenic Potential of the Most Abundant Phospholipase A<sub>2</sub> from Coral Snake <i>Micrurus dumerilii</i> to Develop Antivenoms |
| title_fullStr | Heterologous Expression and Immunogenic Potential of the Most Abundant Phospholipase A<sub>2</sub> from Coral Snake <i>Micrurus dumerilii</i> to Develop Antivenoms |
| title_full_unstemmed | Heterologous Expression and Immunogenic Potential of the Most Abundant Phospholipase A<sub>2</sub> from Coral Snake <i>Micrurus dumerilii</i> to Develop Antivenoms |
| title_short | Heterologous Expression and Immunogenic Potential of the Most Abundant Phospholipase A<sub>2</sub> from Coral Snake <i>Micrurus dumerilii</i> to Develop Antivenoms |
| title_sort | heterologous expression and immunogenic potential of the most abundant phospholipase a sub 2 sub from coral snake i micrurus dumerilii i to develop antivenoms |
| topic | recombinant protein antibodies <i>Micrurus dumerilii</i> coral snake antivenoms phospholipase A<sub>2</sub> |
| url | https://www.mdpi.com/2072-6651/14/12/825 |
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