Analysis of the protein profile of the venoms of snakes Bothrops asper, Bothrocophias myersi and Crotalus durissus from the Colombian Andean Region obtained by RP-HPLC
Snake venoms comprise a highly complex mixture of proteins, and there is also a high interspecific and intraspecific variability in their composition, even in the same region. Our aim was to compare the composition of the venoms of Bothrocophias myersi, Crotalus durissus and Bothrops asper, snakes f...
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| Format: | Article |
| Language: | Spanish |
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Universidad Nacional de Colombia
2021-07-01
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| Series: | Revista Colombiana de Biotecnología |
| Subjects: | |
| Online Access: | https://revistas.unal.edu.co/index.php/biotecnologia/article/view/94211 |
| _version_ | 1819158513292673024 |
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| author | Stefano Scovino Loboguerrero Karen Sarmiento Carlos Galvis Ana Lucía Castiblanco Fabio Aristizabal |
| author_facet | Stefano Scovino Loboguerrero Karen Sarmiento Carlos Galvis Ana Lucía Castiblanco Fabio Aristizabal |
| author_sort | Stefano Scovino Loboguerrero |
| collection | DOAJ |
| description | Snake venoms comprise a highly complex mixture of proteins, and there is also a high interspecific and intraspecific variability in their composition, even in the same region. Our aim was to compare the composition of the venoms of Bothrocophias myersi, Crotalus durissus and Bothrops asper, snakes from the Andean region in Colombia by Reverse-Phase High-Performance Liquid Chromatography (RP-HPLC). The venoms were given to the research group under an agreement with the Fundación Zoológica de Cali. The venoms pool was obtained by manual extraction, lyophilized and refrigerated. The protein found in the venoms was quantified by spectrophotometry using the Bradford and Lowry methods and direct measurement by Nanodrop®. The protein composition was stablished by RP-HPLC, using a Lichosper 100 RP, C18 column (250X4 mm) with a pore size of 5µm, as well as by Sodium Dodecyl Sulfate-Polyacrylamide Gel Electrophoresis (SDS-PAGE). The highest quantity of protein was found in the venom of B. myersi (108,6 mg/mL) followed by C. durissus (78,1 mg/mL) and B. asper (74.1 mg/mL). All venoms showed bands of 15 and 50 KDa by SDS-PAGE; The most important finding is the abundance of PLA2 and svMP in the venom of B. myersi. Chromatographic analyses revealed a very similar venom composition profile, but also certain differences in toxins abundance. We conclude that the process of separating the venom proteins by RP-HPLC and SDS-PAGE are very important as a first step to know the venoms profiles, which in turn could allow medical staff to elucidate the clinical syndrome produced by snakebites. |
| first_indexed | 2024-12-22T16:25:51Z |
| format | Article |
| id | doaj.art-a99d232548324b7c85500b444df35c57 |
| institution | Directory Open Access Journal |
| issn | 0123-3475 1909-8758 |
| language | Spanish |
| last_indexed | 2024-12-22T16:25:51Z |
| publishDate | 2021-07-01 |
| publisher | Universidad Nacional de Colombia |
| record_format | Article |
| series | Revista Colombiana de Biotecnología |
| spelling | doaj.art-a99d232548324b7c85500b444df35c572022-12-21T18:20:09ZspaUniversidad Nacional de ColombiaRevista Colombiana de Biotecnología0123-34751909-87582021-07-0123110.15446/rev.colomb.biote.v23n1.94211Analysis of the protein profile of the venoms of snakes Bothrops asper, Bothrocophias myersi and Crotalus durissus from the Colombian Andean Region obtained by RP-HPLCStefano Scovino Loboguerrero0Karen Sarmiento 1Carlos Galvis2Ana Lucía Castiblanco 3Fabio Aristizabal4Instituto Biotecnología Universidad Nacional2Departament of Physiological Sciences, Faculty of Medicine, Pontificia Universidad Javeriana. Fundación Zoológica de CaliInstituto Biotecnología Universidad NacionalDepartamento de Farmacia, Universidad Nacional Snake venoms comprise a highly complex mixture of proteins, and there is also a high interspecific and intraspecific variability in their composition, even in the same region. Our aim was to compare the composition of the venoms of Bothrocophias myersi, Crotalus durissus and Bothrops asper, snakes from the Andean region in Colombia by Reverse-Phase High-Performance Liquid Chromatography (RP-HPLC). The venoms were given to the research group under an agreement with the Fundación Zoológica de Cali. The venoms pool was obtained by manual extraction, lyophilized and refrigerated. The protein found in the venoms was quantified by spectrophotometry using the Bradford and Lowry methods and direct measurement by Nanodrop®. The protein composition was stablished by RP-HPLC, using a Lichosper 100 RP, C18 column (250X4 mm) with a pore size of 5µm, as well as by Sodium Dodecyl Sulfate-Polyacrylamide Gel Electrophoresis (SDS-PAGE). The highest quantity of protein was found in the venom of B. myersi (108,6 mg/mL) followed by C. durissus (78,1 mg/mL) and B. asper (74.1 mg/mL). All venoms showed bands of 15 and 50 KDa by SDS-PAGE; The most important finding is the abundance of PLA2 and svMP in the venom of B. myersi. Chromatographic analyses revealed a very similar venom composition profile, but also certain differences in toxins abundance. We conclude that the process of separating the venom proteins by RP-HPLC and SDS-PAGE are very important as a first step to know the venoms profiles, which in turn could allow medical staff to elucidate the clinical syndrome produced by snakebites.https://revistas.unal.edu.co/index.php/biotecnologia/article/view/94211Bothrops asper; Bothrocophias myersi; Crotalus durissus; Venom; Colombia; RP-HPLC |
| spellingShingle | Stefano Scovino Loboguerrero Karen Sarmiento Carlos Galvis Ana Lucía Castiblanco Fabio Aristizabal Analysis of the protein profile of the venoms of snakes Bothrops asper, Bothrocophias myersi and Crotalus durissus from the Colombian Andean Region obtained by RP-HPLC Revista Colombiana de Biotecnología Bothrops asper; Bothrocophias myersi; Crotalus durissus; Venom; Colombia; RP-HPLC |
| title | Analysis of the protein profile of the venoms of snakes Bothrops asper, Bothrocophias myersi and Crotalus durissus from the Colombian Andean Region obtained by RP-HPLC |
| title_full | Analysis of the protein profile of the venoms of snakes Bothrops asper, Bothrocophias myersi and Crotalus durissus from the Colombian Andean Region obtained by RP-HPLC |
| title_fullStr | Analysis of the protein profile of the venoms of snakes Bothrops asper, Bothrocophias myersi and Crotalus durissus from the Colombian Andean Region obtained by RP-HPLC |
| title_full_unstemmed | Analysis of the protein profile of the venoms of snakes Bothrops asper, Bothrocophias myersi and Crotalus durissus from the Colombian Andean Region obtained by RP-HPLC |
| title_short | Analysis of the protein profile of the venoms of snakes Bothrops asper, Bothrocophias myersi and Crotalus durissus from the Colombian Andean Region obtained by RP-HPLC |
| title_sort | analysis of the protein profile of the venoms of snakes bothrops asper bothrocophias myersi and crotalus durissus from the colombian andean region obtained by rp hplc |
| topic | Bothrops asper; Bothrocophias myersi; Crotalus durissus; Venom; Colombia; RP-HPLC |
| url | https://revistas.unal.edu.co/index.php/biotecnologia/article/view/94211 |
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