Natiela Beatriz de Oliveira, Ana Carolina Martins Magalhães, Carlos Bloch, Paulo Sérgio Lacerda Beirão, Anita de Oliveira Silva, Rafael D Melani, Eder Alves Barbosa, Osmindo Rodrigues Pires, Carlos Alberto Schwartz
{"title":"Characterization of the first two toxins isolated from the venom of the ancient scorpion <i>Tityus (Archaeotityus) mattogrossensis</i> (Borelli, 1901).","authors":"Natiela Beatriz de Oliveira, Ana Carolina Martins Magalhães, Carlos Bloch, Paulo Sérgio Lacerda Beirão, Anita de Oliveira Silva, Rafael D Melani, Eder Alves Barbosa, Osmindo Rodrigues Pires, Carlos Alberto Schwartz","doi":"10.1590/1678-9199-JVATITD-2021-0035","DOIUrl":null,"url":null,"abstract":"<p><strong>Background: </strong>Almost all <i>Tityus</i> characterized toxins are from subgenera <i>Atreus</i> and <i>Tityus</i>, there are only a few data about toxins produced by <i>Archaeotityus</i>, an ancient group in <i>Tityus</i> genus.</p><p><strong>Methods: </strong><i>Tityus (Archaeotityus) mattogrossensis</i> crude venom was fractionated by high performance liquid chromatography, the major fractions were tested in a frog sciatic nerve single sucrose-gap technique. Two fractions (Tm1 and Tm2) were isolated, partially sequenced by MALDI-TOF/MS and electrophysiological assayed on HEK293 Nav 1.3, HEK293 Nav 1.6, DUM and DRG cells.</p><p><strong>Results: </strong>The sucrose-gap technique showed neurotoxicity in four fractions. One fraction caused a delay of action potential repolarization and other three caused a reduction in amplitude. An electrophysiological assay showed that Tm1 is active on HEK293 Nav 1.3, HEK293 Nav 1.6, DUM and DRG cells, and Tm2 on HEK293 Nav 1.3 and DRG cells, but not in HEK293 Nav 1.6. In addition, Tm1 and Tm2 did promote a shift to more negative potentials strongly suggesting that both are α-NaScTx.</p><p><strong>Conclusion: </strong>Although <i>Tityus (Archaeotityus) mattogrossensis</i> is considered an ancient group in <i>Tityus</i> genus, the primary structure of Tm1 and Tm2 is more related to <i>Tityus</i> subgenus. The patch clamp electrophysiological tests suggest that Tm1 and Tm2 are NaScTx, and also promoted no shift to more negative potentials, strongly suggesting that both are α-NaScTx. This paper aimed to explore and characterize for the first time toxins from the ancient scorpion <i>Tityus (Archaeotityus) mattogrossensis.</i></p>","PeriodicalId":17565,"journal":{"name":"Journal of Venomous Animals and Toxins Including Tropical Diseases","volume":null,"pages":null},"PeriodicalIF":1.8000,"publicationDate":"2021-12-13","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC8670738/pdf/","citationCount":"1","resultStr":null,"platform":"Semanticscholar","paperid":null,"PeriodicalName":"Journal of Venomous Animals and Toxins Including Tropical Diseases","FirstCategoryId":"3","ListUrlMain":"https://doi.org/10.1590/1678-9199-JVATITD-2021-0035","RegionNum":3,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":null,"EPubDate":"2021/1/1 0:00:00","PubModel":"eCollection","JCR":"Q4","JCRName":"TOXICOLOGY","Score":null,"Total":0}
引用次数: 1
Abstract
Background: Almost all Tityus characterized toxins are from subgenera Atreus and Tityus, there are only a few data about toxins produced by Archaeotityus, an ancient group in Tityus genus.
Methods: Tityus (Archaeotityus) mattogrossensis crude venom was fractionated by high performance liquid chromatography, the major fractions were tested in a frog sciatic nerve single sucrose-gap technique. Two fractions (Tm1 and Tm2) were isolated, partially sequenced by MALDI-TOF/MS and electrophysiological assayed on HEK293 Nav 1.3, HEK293 Nav 1.6, DUM and DRG cells.
Results: The sucrose-gap technique showed neurotoxicity in four fractions. One fraction caused a delay of action potential repolarization and other three caused a reduction in amplitude. An electrophysiological assay showed that Tm1 is active on HEK293 Nav 1.3, HEK293 Nav 1.6, DUM and DRG cells, and Tm2 on HEK293 Nav 1.3 and DRG cells, but not in HEK293 Nav 1.6. In addition, Tm1 and Tm2 did promote a shift to more negative potentials strongly suggesting that both are α-NaScTx.
Conclusion: Although Tityus (Archaeotityus) mattogrossensis is considered an ancient group in Tityus genus, the primary structure of Tm1 and Tm2 is more related to Tityus subgenus. The patch clamp electrophysiological tests suggest that Tm1 and Tm2 are NaScTx, and also promoted no shift to more negative potentials, strongly suggesting that both are α-NaScTx. This paper aimed to explore and characterize for the first time toxins from the ancient scorpion Tityus (Archaeotityus) mattogrossensis.
背景:Tityus属的毒素几乎全部来自于Atreus亚属和Tityus亚属,关于tiyus属古类群Archaeotityus产生的毒素资料较少。方法:采用高效液相色谱法对粗毒进行分离,主要组分采用蛙坐骨神经单蔗糖间隙法测定。分离Tm1和Tm2两个部分,用MALDI-TOF/MS进行部分测序,并对HEK293 Nav 1.3、HEK293 Nav 1.6、DUM和DRG细胞进行电生理检测。结果:蔗糖间隙法在4个部位均显示神经毒性。一个片段引起动作电位复极的延迟,另外三个片段引起幅度的减少。电生理实验表明,Tm1对HEK293 Nav 1.3、HEK293 Nav 1.6、DUM和DRG细胞有活性,Tm2对HEK293 Nav 1.3和DRG细胞有活性,但对HEK293 Nav 1.6细胞无活性。此外,Tm1和Tm2确实促进了向更多负电位的转变,这强烈表明它们都是α-NaScTx。结论:虽然Tityus (Archaeotityus) mattogrossensis被认为是Tityus属的一个古老类群,但Tm1和Tm2的初级结构与Tityus亚属的关系更密切。膜片钳电生理测试提示Tm1和Tm2为NaScTx,且未促进更多负电位的移位,强烈提示两者均为α-NaScTx。本文首次对古蝎(Archaeotityus) mattogrossensis的毒素进行了研究和鉴定。
期刊介绍:
Journal of Venomous Animals and Toxins including Tropical Diseases (JVATiTD) is a non-commercial academic open access publication dedicated to research on all aspects of toxinology, venomous animals and tropical diseases. Its interdisciplinary content includes original scientific articles covering research on toxins derived from animals, plants and microorganisms. Topics of interest include, but are not limited to:systematics and morphology of venomous animals;physiology, biochemistry, pharmacology and immunology of toxins;epidemiology, clinical aspects and treatment of envenoming by different animals, plants and microorganisms;development and evaluation of antivenoms and toxin-derivative products;epidemiology, clinical aspects and treatment of tropical diseases (caused by virus, bacteria, algae, fungi and parasites) including the neglected tropical diseases (NTDs) defined by the World Health Organization.