de Oliveira Natiela Beatriz, Magalhães Ana Carolina Martins, Bloch Carlos, Beirão Paulo Sérgio Lacerda, Silva Anita de Oliveira, Melani Rafael D, Barbosa Eder Alves, Pires Osmindo Rodrigues, Schwartz Carlos Alberto
Laboratory of Toxinology, Department of Physiological Sciences, Institute of Biology, University of Brasilia (UnB), Brasília, DF, Brazil.
EMPRABA Center of Genetic Resources and Biotechnology, Brazilian Agricultural Research Corporation, Brasília, DF, Brazil.
J Venom Anim Toxins Incl Trop Dis. 2021 Dec 13;27:e20210035. doi: 10.1590/1678-9199-JVATITD-2021-0035. eCollection 2021.
Almost all characterized toxins are from subgenera and , there are only a few data about toxins produced by , an ancient group in genus.
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.
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.
Although is considered an ancient group in genus, the primary structure of Tm1 and Tm2 is more related to 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
几乎所有已鉴定的毒素都来自亚属和,关于属中一个古老类群产生的毒素的数据很少。
粗毒液通过高效液相色谱法进行分离,主要馏分在青蛙坐骨神经单蔗糖间隙技术中进行测试。分离出两个馏分(Tm1和Tm2),通过基质辅助激光解吸电离飞行时间质谱(MALDI-TOF/MS)进行部分测序,并在HEK293 Nav 1.3、HEK293 Nav 1.6、背根神经节(DUM)和背根神经节(DRG)细胞上进行电生理分析。
蔗糖间隙技术显示四个馏分具有神经毒性。一个馏分导致动作电位复极化延迟,其他三个馏分导致幅度降低。电生理分析表明,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无活性。此外,Tm1和Tm2确实强烈促进了向更负电位的转变,强烈表明两者都是α-神经毒素(α-NaScTx)。
尽管被认为是属中的一个古老类群,但Tm1和Tm2的一级结构与亚属的关系更为密切。膜片钳电生理测试表明,Tm1和Tm2是神经毒素(NaScTx),并且也没有促进向更负电位的转变,强烈表明两者都是α-神经毒素(α-NaScTx)。本文旨在首次探索和鉴定来自古老蝎子的毒素