Purification and characterization of a hyaluronidase from venom of the spider Vitalius dubius (Araneae, Theraphosidae)
BackgroundVenom hyaluronidase (Hyase) contributes to the diffusion of venom from the inoculation site. In this work, we purified and characterized Hyase from the venom of Vitalius dubius (Araneae, Theraphosidae), a large theraphosid found in southeastern Brazil. Venom obtained by electrical stimulat...
Published in: | Journal of Venomous Animals and Toxins including Tropical Diseases |
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ftdoajarticles:oai:doaj.org/article:e9c19636dd6944fa83c0349f116b5616 2023-05-15T15:12:12+02:00 Purification and characterization of a hyaluronidase from venom of the spider Vitalius dubius (Araneae, Theraphosidae) Rafael Sutti Mariana Leite Tamascia Stephen Hyslop Thomaz Augusto Alves Rocha-e-Silva 2014-02-01T00:00:00Z https://doi.org/10.1186/1678-9199-20-2 https://doaj.org/article/e9c19636dd6944fa83c0349f116b5616 EN eng SciELO http://www.scielo.br/scielo.php?script=sci_arttext&pid=S1678-91992014000200312&lng=en&tlng=en https://doaj.org/toc/1678-9199 1678-9199 doi:10.1186/1678-9199-20-2 https://doaj.org/article/e9c19636dd6944fa83c0349f116b5616 Journal of Venomous Animals and Toxins including Tropical Diseases, Vol 20, Iss 0 (2014) Hyaluronidase Venom Purification Vitalius dubius Arctic medicine. Tropical medicine RC955-962 Toxicology. Poisons RA1190-1270 Zoology QL1-991 article 2014 ftdoajarticles https://doi.org/10.1186/1678-9199-20-2 2022-12-30T23:53:52Z BackgroundVenom hyaluronidase (Hyase) contributes to the diffusion of venom from the inoculation site. In this work, we purified and characterized Hyase from the venom of Vitalius dubius (Araneae, Theraphosidae), a large theraphosid found in southeastern Brazil. Venom obtained by electrical stimulation of adult male and female V. dubius was initially fractionated by gel filtration on a Superdex® 75 column. Active fractions were pooled and applied to a heparin-sepharose affinity column. The proteins were eluted with a linear NaCl gradient.ResultsActive fractions were pooled and assessed for purity by SDS-PAGE and RP-HPLC. The physicochemical tests included optimum pH, heat stability, presence of isoforms, neutralization by flavonoids and assessment of commercial antivenoms. Hyase was purified and presented a specific activity of 148 turbidity-reducing units (TRU)/mg (venom: 36 TRU/mg; purification factor of ~4). Hyase displayed a molecular mass of 43 kDa by SDS-PAGE. Zymography in hyaluronic-acid-containing gels indicated an absence of enzyme isoforms. The optimum pH was 4-5, with highest activity at 37°C. Hyase was stable up to 60°C; but its activity was lost at higher temperatures and maintained after several freeze-thaw cycles. The NaCl concentration (up to 1 M) did not influence activity. Hyase had greater action towards hyaluronic acid compared to chondroitin sulfate, and was completely neutralized by polyvalent antiarachnid sera, but not by caterpillar, scorpion or snakes antivenoms.ConclusionThe neutralization by arachnid but not scorpion antivenom indicates that this enzyme shares antigenic epitopes with similar enzymes in other spider venoms. The biochemical properties of this Hyase are comparable to others described. Article in Journal/Newspaper Arctic Directory of Open Access Journals: DOAJ Articles Arctic Journal of Venomous Animals and Toxins including Tropical Diseases 20 1 2 |
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Directory of Open Access Journals: DOAJ Articles |
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English |
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Hyaluronidase Venom Purification Vitalius dubius Arctic medicine. Tropical medicine RC955-962 Toxicology. Poisons RA1190-1270 Zoology QL1-991 |
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Hyaluronidase Venom Purification Vitalius dubius Arctic medicine. Tropical medicine RC955-962 Toxicology. Poisons RA1190-1270 Zoology QL1-991 Rafael Sutti Mariana Leite Tamascia Stephen Hyslop Thomaz Augusto Alves Rocha-e-Silva Purification and characterization of a hyaluronidase from venom of the spider Vitalius dubius (Araneae, Theraphosidae) |
topic_facet |
Hyaluronidase Venom Purification Vitalius dubius Arctic medicine. Tropical medicine RC955-962 Toxicology. Poisons RA1190-1270 Zoology QL1-991 |
description |
BackgroundVenom hyaluronidase (Hyase) contributes to the diffusion of venom from the inoculation site. In this work, we purified and characterized Hyase from the venom of Vitalius dubius (Araneae, Theraphosidae), a large theraphosid found in southeastern Brazil. Venom obtained by electrical stimulation of adult male and female V. dubius was initially fractionated by gel filtration on a Superdex® 75 column. Active fractions were pooled and applied to a heparin-sepharose affinity column. The proteins were eluted with a linear NaCl gradient.ResultsActive fractions were pooled and assessed for purity by SDS-PAGE and RP-HPLC. The physicochemical tests included optimum pH, heat stability, presence of isoforms, neutralization by flavonoids and assessment of commercial antivenoms. Hyase was purified and presented a specific activity of 148 turbidity-reducing units (TRU)/mg (venom: 36 TRU/mg; purification factor of ~4). Hyase displayed a molecular mass of 43 kDa by SDS-PAGE. Zymography in hyaluronic-acid-containing gels indicated an absence of enzyme isoforms. The optimum pH was 4-5, with highest activity at 37°C. Hyase was stable up to 60°C; but its activity was lost at higher temperatures and maintained after several freeze-thaw cycles. The NaCl concentration (up to 1 M) did not influence activity. Hyase had greater action towards hyaluronic acid compared to chondroitin sulfate, and was completely neutralized by polyvalent antiarachnid sera, but not by caterpillar, scorpion or snakes antivenoms.ConclusionThe neutralization by arachnid but not scorpion antivenom indicates that this enzyme shares antigenic epitopes with similar enzymes in other spider venoms. The biochemical properties of this Hyase are comparable to others described. |
format |
Article in Journal/Newspaper |
author |
Rafael Sutti Mariana Leite Tamascia Stephen Hyslop Thomaz Augusto Alves Rocha-e-Silva |
author_facet |
Rafael Sutti Mariana Leite Tamascia Stephen Hyslop Thomaz Augusto Alves Rocha-e-Silva |
author_sort |
Rafael Sutti |
title |
Purification and characterization of a hyaluronidase from venom of the spider Vitalius dubius (Araneae, Theraphosidae) |
title_short |
Purification and characterization of a hyaluronidase from venom of the spider Vitalius dubius (Araneae, Theraphosidae) |
title_full |
Purification and characterization of a hyaluronidase from venom of the spider Vitalius dubius (Araneae, Theraphosidae) |
title_fullStr |
Purification and characterization of a hyaluronidase from venom of the spider Vitalius dubius (Araneae, Theraphosidae) |
title_full_unstemmed |
Purification and characterization of a hyaluronidase from venom of the spider Vitalius dubius (Araneae, Theraphosidae) |
title_sort |
purification and characterization of a hyaluronidase from venom of the spider vitalius dubius (araneae, theraphosidae) |
publisher |
SciELO |
publishDate |
2014 |
url |
https://doi.org/10.1186/1678-9199-20-2 https://doaj.org/article/e9c19636dd6944fa83c0349f116b5616 |
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Arctic |
geographic_facet |
Arctic |
genre |
Arctic |
genre_facet |
Arctic |
op_source |
Journal of Venomous Animals and Toxins including Tropical Diseases, Vol 20, Iss 0 (2014) |
op_relation |
http://www.scielo.br/scielo.php?script=sci_arttext&pid=S1678-91992014000200312&lng=en&tlng=en https://doaj.org/toc/1678-9199 1678-9199 doi:10.1186/1678-9199-20-2 https://doaj.org/article/e9c19636dd6944fa83c0349f116b5616 |
op_doi |
https://doi.org/10.1186/1678-9199-20-2 |
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Journal of Venomous Animals and Toxins including Tropical Diseases |
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20 |
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2 |
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1766342924282363904 |