The Venom of <i>Vipera ammodytes ammodytes</i>: Proteomics, Neurotoxic Effect and Neutralization by Antivenom

Deep proteomic analyses identified, in total, 159 master proteins (with 1% FDR and 2 unique peptides) from 26 protein families in the venom of <i>Vipera ammodytes ammodytes (Vaa).</i> Data are available via ProteomeXchange with the identifier PXD056495. The relative abundance of PLA2s is...

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Main Authors: Saša R. Ivanović, Dina Rešetar Maslov, Ivana Rubić, Vladimir Mrljak, Irena Živković, Nevena Borozan, Jelica Grujić-Milanović, Sunčica Borozan
Format: Article
Language:English
Published: MDPI AG 2024-11-01
Series:Veterinary Sciences
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Online Access:https://www.mdpi.com/2306-7381/11/12/605
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author Saša R. Ivanović
Dina Rešetar Maslov
Ivana Rubić
Vladimir Mrljak
Irena Živković
Nevena Borozan
Jelica Grujić-Milanović
Sunčica Borozan
author_facet Saša R. Ivanović
Dina Rešetar Maslov
Ivana Rubić
Vladimir Mrljak
Irena Živković
Nevena Borozan
Jelica Grujić-Milanović
Sunčica Borozan
author_sort Saša R. Ivanović
collection DOAJ
description Deep proteomic analyses identified, in total, 159 master proteins (with 1% FDR and 2 unique peptides) from 26 protein families in the venom of <i>Vipera ammodytes ammodytes (Vaa).</i> Data are available via ProteomeXchange with the identifier PXD056495. The relative abundance of PLA2s is 11.60% of the crude venom, of which 4.35% are neurotoxic Ammodytoxins (Atxs). The neurotoxicity of the venom of <i>Vaa</i> and the neutralizing effect of the antivenom were tested on the neuromuscular preparation of the diaphragm (NPD) of rats. The activity of PLA2 in the venom of <i>Vaa</i> and its neutralization by the antivenom were determined under in vitro conditions. The <i>Vaa</i> venom leads to a progressive decrease in NPD contractions. We administered pre-incubated venom/antivenom mixtures at various ratios of 1:2, 1:10 and 1:20 (<i>w</i>/<i>w</i>) and observed the effects of these mixtures on NPD contractions. The results show that the mean effective time (ET<sub>50</sub>) for NPD contractions with the 1:20 mixture is highly significantly different (<i>p</i> < 0.001) from the ET<sub>50</sub> for the venom and the ET<sub>50</sub> for the 1:2 and 1:10 mixture ratios. We also found a highly significant (<i>p</i> < 0.001) reduction in Na<sup>+</sup>/K<sup>+</sup>-ATPase activity in the NPD under the influence of the venom. The reduction in the activity of this enzyme was reversible by the antivenom. Under in vitro conditions, we have achieved the complete neutralization of PLA2 by the antivenom. In conclusion, the antivenom abolished the venom-induced progressive decrease in NPD contractions in a concentration-dependent manner. Antivenom with approximately the same mass proportion almost completely restores Na<sup>+</sup>/K<sup>+</sup>-ATPase activity in the NPD and completely neutralizes the PLA2 activity of the venom in vitro.
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spelling doaj-art-68fdf5ead3304ee4888a1177787790b02025-08-20T02:01:24ZengMDPI AGVeterinary Sciences2306-73812024-11-01111260510.3390/vetsci11120605The Venom of <i>Vipera ammodytes ammodytes</i>: Proteomics, Neurotoxic Effect and Neutralization by AntivenomSaša R. Ivanović0Dina Rešetar Maslov1Ivana Rubić2Vladimir Mrljak3Irena Živković4Nevena Borozan5Jelica Grujić-Milanović6Sunčica Borozan7Department of Pharmacology and Toxicology, Faculty of Veterinary Medicine, University of Belgrade, Bulevar Oslobođenja 18, 11000 Belgrade, SerbiaLaboratory of Proteomics, Clinic for Internal Diseases, Faculty of Veterinary Medicine, University of Zagreb, Heinzelova 55, 10000 Zagreb, CroatiaLaboratory of Proteomics, Clinic for Internal Diseases, Faculty of Veterinary Medicine, University of Zagreb, Heinzelova 55, 10000 Zagreb, CroatiaLaboratory of Proteomics, Clinic for Internal Diseases, Faculty of Veterinary Medicine, University of Zagreb, Heinzelova 55, 10000 Zagreb, CroatiaInstitute of Virology, Vaccines and Sera “Torlak”, Vojvode Stepe 458, 11000 Belgrade, SerbiaFaculty of Medicine, University of Belgrade, Dr Subotića 1, 11000 Belgrade, SerbiaDepartment of Cardiovascular Research, Institute for Medical Research, National Institute of the Republic of Serbia, University of Belgrade, 11000 Belgrade, SerbiaDepartment of Chemistry, Faculty of Veterinary Medicine, University of Belgrade, Bulevar Oslobođenja 18, 11000 Belgrade, SerbiaDeep proteomic analyses identified, in total, 159 master proteins (with 1% FDR and 2 unique peptides) from 26 protein families in the venom of <i>Vipera ammodytes ammodytes (Vaa).</i> Data are available via ProteomeXchange with the identifier PXD056495. The relative abundance of PLA2s is 11.60% of the crude venom, of which 4.35% are neurotoxic Ammodytoxins (Atxs). The neurotoxicity of the venom of <i>Vaa</i> and the neutralizing effect of the antivenom were tested on the neuromuscular preparation of the diaphragm (NPD) of rats. The activity of PLA2 in the venom of <i>Vaa</i> and its neutralization by the antivenom were determined under in vitro conditions. The <i>Vaa</i> venom leads to a progressive decrease in NPD contractions. We administered pre-incubated venom/antivenom mixtures at various ratios of 1:2, 1:10 and 1:20 (<i>w</i>/<i>w</i>) and observed the effects of these mixtures on NPD contractions. The results show that the mean effective time (ET<sub>50</sub>) for NPD contractions with the 1:20 mixture is highly significantly different (<i>p</i> < 0.001) from the ET<sub>50</sub> for the venom and the ET<sub>50</sub> for the 1:2 and 1:10 mixture ratios. We also found a highly significant (<i>p</i> < 0.001) reduction in Na<sup>+</sup>/K<sup>+</sup>-ATPase activity in the NPD under the influence of the venom. The reduction in the activity of this enzyme was reversible by the antivenom. Under in vitro conditions, we have achieved the complete neutralization of PLA2 by the antivenom. In conclusion, the antivenom abolished the venom-induced progressive decrease in NPD contractions in a concentration-dependent manner. Antivenom with approximately the same mass proportion almost completely restores Na<sup>+</sup>/K<sup>+</sup>-ATPase activity in the NPD and completely neutralizes the PLA2 activity of the venom in vitro.https://www.mdpi.com/2306-7381/11/12/605proteomicsvenom<i>Vipera ammodytes ammodytes</i>neurotoxicitydiaphragmNa<sup>+</sup>/K<sup>+</sup>-ATPase
spellingShingle Saša R. Ivanović
Dina Rešetar Maslov
Ivana Rubić
Vladimir Mrljak
Irena Živković
Nevena Borozan
Jelica Grujić-Milanović
Sunčica Borozan
The Venom of <i>Vipera ammodytes ammodytes</i>: Proteomics, Neurotoxic Effect and Neutralization by Antivenom
Veterinary Sciences
proteomics
venom
<i>Vipera ammodytes ammodytes</i>
neurotoxicity
diaphragm
Na<sup>+</sup>/K<sup>+</sup>-ATPase
title The Venom of <i>Vipera ammodytes ammodytes</i>: Proteomics, Neurotoxic Effect and Neutralization by Antivenom
title_full The Venom of <i>Vipera ammodytes ammodytes</i>: Proteomics, Neurotoxic Effect and Neutralization by Antivenom
title_fullStr The Venom of <i>Vipera ammodytes ammodytes</i>: Proteomics, Neurotoxic Effect and Neutralization by Antivenom
title_full_unstemmed The Venom of <i>Vipera ammodytes ammodytes</i>: Proteomics, Neurotoxic Effect and Neutralization by Antivenom
title_short The Venom of <i>Vipera ammodytes ammodytes</i>: Proteomics, Neurotoxic Effect and Neutralization by Antivenom
title_sort venom of i vipera ammodytes ammodytes i proteomics neurotoxic effect and neutralization by antivenom
topic proteomics
venom
<i>Vipera ammodytes ammodytes</i>
neurotoxicity
diaphragm
Na<sup>+</sup>/K<sup>+</sup>-ATPase
url https://www.mdpi.com/2306-7381/11/12/605
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