Crotoxin B: Heterologous Expression, Protein Folding, Immunogenic Properties, and Irregular Presence in Crotalid Venoms
Crotoxin complex CA/CB and crotamine are the main toxins associated with <i>Crotalus</i> envenomation besides the enzymatic activities of phospholipases (PLA<sub>2</sub>) and proteases. The neutralization at least of the crotoxin complex by neutralizing the subunit B could be...
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| Format: | Article |
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MDPI AG
2022-05-01
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| Series: | Toxins |
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| Online Access: | https://www.mdpi.com/2072-6651/14/6/382 |
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| author | Miguel Angel Mejía-Sánchez Herlinda Clement Ligia Luz Corrales-García Timoteo Olamendi-Portugal Alejandro Carbajal Gerardo Corzo |
| author_facet | Miguel Angel Mejía-Sánchez Herlinda Clement Ligia Luz Corrales-García Timoteo Olamendi-Portugal Alejandro Carbajal Gerardo Corzo |
| author_sort | Miguel Angel Mejía-Sánchez |
| collection | DOAJ |
| description | Crotoxin complex CA/CB and crotamine are the main toxins associated with <i>Crotalus</i> envenomation besides the enzymatic activities of phospholipases (PLA<sub>2</sub>) and proteases. The neutralization at least of the crotoxin complex by neutralizing the subunit B could be a key in the production process of antivenoms against crotalids. Therefore, in this work, a Crotoxin B was recombinantly expressed to evaluate its capacity as an immunogen and its ability to produce neutralizing antibodies against crotalid venoms. A Crotoxin B transcript from <i>Crotalus tzabcan</i> was cloned into a pCR<sup>®</sup>2.1-TOPO vector (Invitrogen, Waltham, MA, USA) and subsequently expressed heterologously in bacteria. HisrCrotoxin B was extracted from inclusion bodies and refolded in vitro. The secondary structure of HisrCrotoxin B was comparable to the secondary structure of the native Crotoxin B, and it has PLA<sub>2</sub> activity as the native Crotoxin B. HisrCrotoxin B was used to immunize rabbits, and the obtained antibodies partially inhibited the activity of PLA<sub>2</sub> from <i>C. tzabcan</i>. The anti-HisrCrotoxin B antibodies neutralized the native Crotoxin B and the whole venoms from <i>C. tzabcan, C. s. salvini</i>, and <i>C. mictlantecuhtli.</i> Additionally, anti-HisrCrotoxin B antibodies recognized native Crotoxin B from different <i>Crotalus</i> species, and they could discriminate venom in species with high or low levels of or absence of Crotoxin B. |
| first_indexed | 2024-03-09T22:19:46Z |
| format | Article |
| id | doaj.art-feb841ebbfcf45e6af41c7455a0765c7 |
| institution | Directory Open Access Journal |
| issn | 2072-6651 |
| language | English |
| last_indexed | 2024-03-09T22:19:46Z |
| publishDate | 2022-05-01 |
| publisher | MDPI AG |
| record_format | Article |
| series | Toxins |
| spelling | doaj.art-feb841ebbfcf45e6af41c7455a0765c72023-11-23T19:16:59ZengMDPI AGToxins2072-66512022-05-0114638210.3390/toxins14060382Crotoxin B: Heterologous Expression, Protein Folding, Immunogenic Properties, and Irregular Presence in Crotalid VenomsMiguel Angel Mejía-Sánchez0Herlinda Clement1Ligia Luz Corrales-García2Timoteo Olamendi-Portugal3Alejandro Carbajal4Gerardo Corzo5Departamento de Medicina Molecular y Bioprocesos, Instituto de Biotecnologia, Universidad Nacional Autónoma de México, Avenida Universidad, 2001, Apartado Postal 510-3, Cuernavaca 62210, MexicoDepartamento de Medicina Molecular y Bioprocesos, Instituto de Biotecnologia, Universidad Nacional Autónoma de México, Avenida Universidad, 2001, Apartado Postal 510-3, Cuernavaca 62210, MexicoDepartamento de Medicina Molecular y Bioprocesos, Instituto de Biotecnologia, Universidad Nacional Autónoma de México, Avenida Universidad, 2001, Apartado Postal 510-3, Cuernavaca 62210, MexicoDepartamento de Medicina Molecular y Bioprocesos, Instituto de Biotecnologia, Universidad Nacional Autónoma de México, Avenida Universidad, 2001, Apartado Postal 510-3, Cuernavaca 62210, MexicoCentro de Investigaciones Biológicas del Noroeste, S.C. Instituto Politécnico Nacional s/n, Playa Palo de Santa Rita Sur, La Paz 23096, MexicoDepartamento de Medicina Molecular y Bioprocesos, Instituto de Biotecnologia, Universidad Nacional Autónoma de México, Avenida Universidad, 2001, Apartado Postal 510-3, Cuernavaca 62210, MexicoCrotoxin complex CA/CB and crotamine are the main toxins associated with <i>Crotalus</i> envenomation besides the enzymatic activities of phospholipases (PLA<sub>2</sub>) and proteases. The neutralization at least of the crotoxin complex by neutralizing the subunit B could be a key in the production process of antivenoms against crotalids. Therefore, in this work, a Crotoxin B was recombinantly expressed to evaluate its capacity as an immunogen and its ability to produce neutralizing antibodies against crotalid venoms. A Crotoxin B transcript from <i>Crotalus tzabcan</i> was cloned into a pCR<sup>®</sup>2.1-TOPO vector (Invitrogen, Waltham, MA, USA) and subsequently expressed heterologously in bacteria. HisrCrotoxin B was extracted from inclusion bodies and refolded in vitro. The secondary structure of HisrCrotoxin B was comparable to the secondary structure of the native Crotoxin B, and it has PLA<sub>2</sub> activity as the native Crotoxin B. HisrCrotoxin B was used to immunize rabbits, and the obtained antibodies partially inhibited the activity of PLA<sub>2</sub> from <i>C. tzabcan</i>. The anti-HisrCrotoxin B antibodies neutralized the native Crotoxin B and the whole venoms from <i>C. tzabcan, C. s. salvini</i>, and <i>C. mictlantecuhtli.</i> Additionally, anti-HisrCrotoxin B antibodies recognized native Crotoxin B from different <i>Crotalus</i> species, and they could discriminate venom in species with high or low levels of or absence of Crotoxin B.https://www.mdpi.com/2072-6651/14/6/382antibodies<i>Crotalus tzabcan</i>phospholipaseprotein expressionsnakevenom |
| spellingShingle | Miguel Angel Mejía-Sánchez Herlinda Clement Ligia Luz Corrales-García Timoteo Olamendi-Portugal Alejandro Carbajal Gerardo Corzo Crotoxin B: Heterologous Expression, Protein Folding, Immunogenic Properties, and Irregular Presence in Crotalid Venoms Toxins antibodies <i>Crotalus tzabcan</i> phospholipase protein expression snake venom |
| title | Crotoxin B: Heterologous Expression, Protein Folding, Immunogenic Properties, and Irregular Presence in Crotalid Venoms |
| title_full | Crotoxin B: Heterologous Expression, Protein Folding, Immunogenic Properties, and Irregular Presence in Crotalid Venoms |
| title_fullStr | Crotoxin B: Heterologous Expression, Protein Folding, Immunogenic Properties, and Irregular Presence in Crotalid Venoms |
| title_full_unstemmed | Crotoxin B: Heterologous Expression, Protein Folding, Immunogenic Properties, and Irregular Presence in Crotalid Venoms |
| title_short | Crotoxin B: Heterologous Expression, Protein Folding, Immunogenic Properties, and Irregular Presence in Crotalid Venoms |
| title_sort | crotoxin b heterologous expression protein folding immunogenic properties and irregular presence in crotalid venoms |
| topic | antibodies <i>Crotalus tzabcan</i> phospholipase protein expression snake venom |
| url | https://www.mdpi.com/2072-6651/14/6/382 |
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