Isolation and functional characterization of a new acidic PLA2 Ba SpII RP4 of the Bothrops alternatus snake venom from Argentina

An acidic protein with phospholipase A2 activity was purified to homogeneity from the venom of the Northeast Argentinian viperid Bothrops alternatus by two chromatographic steps: a conventional gel filtration on Sephadex G-75 and reversed phase on C18 HPLC column. A molecular mass of 14,185.48 Da wa...

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Autores principales: García Denegri, María Emilia, Acosta, Ofelia Cristina, Huancahuire-Vega, Salomón, Martins-de-Souza, Daniel, Marangoni, Sergio, Maruñak, Silvana Licia, Teibler, Gladys Pamela, Leiva, Laura Cristina Ana, Ponce-Soto, Luis Alberto
Formato: Artículo
Lenguaje:Inglés
Publicado: Elsevier 2025
Materias:
Phospholipase A2
Bothrops alternatus
Acidic Asp49
Amino-acid sequence
Non-myotoxic
Non-lethal
Acceso en línea:http://repositorio.unne.edu.ar/handle/123456789/59358
Aporte de:
RIUNNE - Repositorio Institucional de la Universidad Nacional del Nordeste (UNNE) de Universidad Nacional del Nordeste
id I48-R184-123456789-59358
record_format dspace
spelling I48-R184-123456789-593582025-12-18T18:03:17Z Isolation and functional characterization of a new acidic PLA2 Ba SpII RP4 of the Bothrops alternatus snake venom from Argentina García Denegri, María Emilia Acosta, Ofelia Cristina Huancahuire-Vega, Salomón Martins-de-Souza, Daniel Marangoni, Sergio Maruñak, Silvana Licia Teibler, Gladys Pamela Leiva, Laura Cristina Ana Ponce-Soto, Luis Alberto Phospholipase A2 Bothrops alternatus Acidic Asp49 Amino-acid sequence Non-myotoxic Non-lethal An acidic protein with phospholipase A2 activity was purified to homogeneity from the venom of the Northeast Argentinian viperid Bothrops alternatus by two chromatographic steps: a conventional gel filtration on Sephadex G-75 and reversed phase on C18 HPLC column. A molecular mass of 14,185.48 Da was determined by mass spectrometry, displaying a homodimer conformation. The kinetic assay demonstrated a catalytically active phospholipase A2 in correspondence with Asp49 PLA2 group. The enzyme designated Ba SpII RP4 contains an amino acid composition of 121 residues and a calculated theoretical pI value of 4.88. Amino acid sequence alignments with other Bothrops PLA2 revealed a high degree of homology sequence (90–56%). Ba SpII RP4 did not show myotoxic activity upon muscular fibers at doses up to 100 µg via intramuscular injection or lethal response when it was intraperitoneally injected at the highest dose of 200 µg. This toxin generates slight biological activities like paw edema inflammation and a delay in the clotting time, although Ba SpII RP4 exhibited catalytic activity. The primary amino acid sequence, determined by quadruple-time of flight (Q-TOF) hybrid mass spectrometer Q-TOF Ultima from Micromass (Manchester, UK) equipped with a nano Zspray source operating in a positive ion mode and tandem mass spectrum, an ESI/MS mass spectrum (TOF MS mode) “de novo amino acid sequencing,” also provides more database about the small group of the non-myotoxic PLA2s isolated up to the present. 2025-12-15T11:31:20Z 2025-12-15T11:31:20Z 2010-08 Artículo García Denegri, María Emilia, et al., 2010. Isolation and functional characterization of a new acidic PLA2 Ba SpII RP4 of the Bothrops alternatus snake venom from Argentina. Toxicon. Oxford: Elsevier, vol. 56, no. 1, p. 64-74. E-ISSN 1879-3150. 0041-0101 http://repositorio.unne.edu.ar/handle/123456789/59358 eng https://doi.org/10.1016/j.toxicon.2010.02.031 openAccess http://creativecommons.org/licenses/by-nc-nd/2.5/ar/ application/pdf p. 64-74 application/pdf Elsevier Toxicon, 2010, vol. 56, no. 1, p. 64-74.
institution Universidad Nacional del Nordeste
institution_str I-48
repository_str R-184
collection RIUNNE - Repositorio Institucional de la Universidad Nacional del Nordeste (UNNE)
language Inglés
topic Phospholipase A2
Bothrops alternatus
Acidic Asp49
Amino-acid sequence
Non-myotoxic
Non-lethal
spellingShingle Phospholipase A2
Bothrops alternatus
Acidic Asp49
Amino-acid sequence
Non-myotoxic
Non-lethal
García Denegri, María Emilia
Acosta, Ofelia Cristina
Huancahuire-Vega, Salomón
Martins-de-Souza, Daniel
Marangoni, Sergio
Maruñak, Silvana Licia
Teibler, Gladys Pamela
Leiva, Laura Cristina Ana
Ponce-Soto, Luis Alberto
Isolation and functional characterization of a new acidic PLA2 Ba SpII RP4 of the Bothrops alternatus snake venom from Argentina
topic_facet Phospholipase A2
Bothrops alternatus
Acidic Asp49
Amino-acid sequence
Non-myotoxic
Non-lethal
description An acidic protein with phospholipase A2 activity was purified to homogeneity from the venom of the Northeast Argentinian viperid Bothrops alternatus by two chromatographic steps: a conventional gel filtration on Sephadex G-75 and reversed phase on C18 HPLC column. A molecular mass of 14,185.48 Da was determined by mass spectrometry, displaying a homodimer conformation. The kinetic assay demonstrated a catalytically active phospholipase A2 in correspondence with Asp49 PLA2 group. The enzyme designated Ba SpII RP4 contains an amino acid composition of 121 residues and a calculated theoretical pI value of 4.88. Amino acid sequence alignments with other Bothrops PLA2 revealed a high degree of homology sequence (90–56%). Ba SpII RP4 did not show myotoxic activity upon muscular fibers at doses up to 100 µg via intramuscular injection or lethal response when it was intraperitoneally injected at the highest dose of 200 µg. This toxin generates slight biological activities like paw edema inflammation and a delay in the clotting time, although Ba SpII RP4 exhibited catalytic activity. The primary amino acid sequence, determined by quadruple-time of flight (Q-TOF) hybrid mass spectrometer Q-TOF Ultima from Micromass (Manchester, UK) equipped with a nano Zspray source operating in a positive ion mode and tandem mass spectrum, an ESI/MS mass spectrum (TOF MS mode) “de novo amino acid sequencing,” also provides more database about the small group of the non-myotoxic PLA2s isolated up to the present.
format Artículo
author García Denegri, María Emilia
Acosta, Ofelia Cristina
Huancahuire-Vega, Salomón
Martins-de-Souza, Daniel
Marangoni, Sergio
Maruñak, Silvana Licia
Teibler, Gladys Pamela
Leiva, Laura Cristina Ana
Ponce-Soto, Luis Alberto
author_facet García Denegri, María Emilia
Acosta, Ofelia Cristina
Huancahuire-Vega, Salomón
Martins-de-Souza, Daniel
Marangoni, Sergio
Maruñak, Silvana Licia
Teibler, Gladys Pamela
Leiva, Laura Cristina Ana
Ponce-Soto, Luis Alberto
author_sort García Denegri, María Emilia
title Isolation and functional characterization of a new acidic PLA2 Ba SpII RP4 of the Bothrops alternatus snake venom from Argentina
title_short Isolation and functional characterization of a new acidic PLA2 Ba SpII RP4 of the Bothrops alternatus snake venom from Argentina
title_full Isolation and functional characterization of a new acidic PLA2 Ba SpII RP4 of the Bothrops alternatus snake venom from Argentina
title_fullStr Isolation and functional characterization of a new acidic PLA2 Ba SpII RP4 of the Bothrops alternatus snake venom from Argentina
title_full_unstemmed Isolation and functional characterization of a new acidic PLA2 Ba SpII RP4 of the Bothrops alternatus snake venom from Argentina
title_sort isolation and functional characterization of a new acidic pla2 ba spii rp4 of the bothrops alternatus snake venom from argentina
publisher Elsevier
publishDate 2025
url http://repositorio.unne.edu.ar/handle/123456789/59358
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