Isolation and identification of a sodium channel-inhibiting protein from eggs of black widow spiders

Jian-jun, LI; Yan, Yizhong; Yu, Hai; Xu, Peng; Zhang, Yiya; Hu, Weijun; Duan, Zhigui; Wang, Xianchun; Liang, Songping

doi:10.1016/j.ijbiomac.2014.01.004

Cited by 15 publications

(18 citation statements)

References 20 publications

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“…In our previous studies, latroeggtoxin-I to latroeggtoxin-IV were purified from the eggs and identified to be toxins [16,17,18]. However, they had not been identified to be toxins by BLAST analysis in the present transcriptome of the eggs, because the existing databases had not collected them as toxins when our current work was performed.…”

Section: Discussionmentioning

confidence: 99%

“…Besides, three high-molecular-mass proteins and one low-molecular-mass peptide, named latroeggtoxin-I to latroeggtoxin-IV, respectively, were purified and characterized from the egg extract. Among them, Latroeggtoxin-I reversibly blocked the neuromuscular transmission in isolated mouse phrenic nerve-hemidiaphragm preparations; latroeggtoxin-II selectively inhibited tetrodotoxin-resistant (TTX-R) sodium channel currents in rat dorsal root ganglion neurons (DRG), without significant effect on the tetrodotoxin-sensitive (TTX-S) sodium channel currents; latroeggtoxin-III showed inhibitory effects on the potassium and calcium channel currents in cockroach dorsal unpaired median (DUM) neurons; and latroeggtoxin-IV was shown to be a broad-spectrum antibacterial peptide [16,17,18]. …”

Section: Introductionmentioning

confidence: 99%

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Transcriptome Analysis to Understand the Toxicity of Latrodectus tredecimguttatus Eggs

Wang

2016

Toxins

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Latrodectus tredecimguttatus is a kind of highly venomous black widow spider, with toxicity coming from not only venomous glands but also other parts of its body as well as newborn spiderlings and eggs. Up to date, although L. tredecimguttatus eggs have been demonstrated to be rich in proteinaceous toxins, there is no systematic investigation on such active components at transcriptome level. In this study, we performed a high-throughput transcriptome sequencing of L. tredecimguttatus eggs with Illumina sequencing technology. As a result, 53,284 protein-coding unigenes were identified, of which 14,185 unigenes produced significant hits in the available databases, including 280 unigenes encoding proteins or peptides homologous to known proteinaceous toxins. GO term and KEGG pathway enrichment analyses of the 280 unigenes showed that 375 GO terms and 18 KEGG pathways were significantly enriched. Functional analysis indicated that these unigene-coded toxins have the bioactivities to degrade tissue proteins, inhibit ion channels, block neuromuscular transmission, provoke anaphylaxis, induce apoptosis and hyperalgesia, etc. No known typical proteinaceous toxins in L. tredecimguttatus venomous glands, such as latrotoxins, were identified, suggesting that the eggs have a different toxicity mechanism from that of the venom. Our present transcriptome analysis not only helps to reveal the gene expression profile and toxicity mechanism of the L. tredecimguttatus eggs, but also provides references for the further related researches.

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Section: Discussionmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

Transcriptome Analysis to Understand the Toxicity of Latrodectus tredecimguttatus Eggs

Wang

2016

Toxins

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“…Furthermore, we purified a protein (named Latroeggtoxin-II) from the egg extract in our previous study. Electrophysiological experiments demonstrated that the protein can selectively inhibit TTX-R Na + channel currents in rat DRG neurons, without significant effect on the TTX-S Na + channel currents [21]. …”

Section: Discussionmentioning

confidence: 99%

Physiological and biochemical characterization of egg extract of black widow spiders to uncover molecular basis of egg toxicity

Yan

Zhang

et al. 2014

biol res

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BackgroundBlack widow spider (L. tredecimguttatus) has toxic components not only in the venomous glands, but also in other parts of the body and its eggs. It is biologically important to investigate the molecular basis of the egg toxicity.ResultsIn the present work, an aqueous extract was prepared from the eggs of the spider and characterized using multiple physiological and biochemical strategies. Gel electrophoresis and mass spectrometry demonstrated that the eggs are rich in high-molecular-mass proteins and the peptides below 5 kDa. The lyophilized extract of the eggs had a protein content of 34.22% and was shown to have a strong toxicity towards mammals and insects. When applied at a concentration of 0.25 mg/mL, the extract could completely block the neuromuscular transmission in mouse isolated phrenic nerve-hemidiaphragm preparations within 12.0 ± 1.5 min. Using whole-cell patch-clamp technique, the egg extract was demonstrated to be able to inhibit the voltage-activated Na+, K+ and Ca2+ currents in rat DRG neurons. In addition, the extract displayed activities of multiple hydrolases. Finally, the molecular basis of the egg toxicity was discussed.ConclusionsThe eggs of black widow spiders are rich in proteinous compounds particularly the high-molecular-mass proteins with different types of biological activity The neurotoxic and other active compounds in the eggs are believed to play important roles in the eggs’ toxic actions.

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“…By using gel filtration combined with ion-exchange chromatography as well as RP-HPLC, two active proteins were purified from the eggs, named Latroeggtoxin-I and Latroeggtoxin-II, respectively. They have been demonstrated to be novel neurotoxins purified from the eggs of black widow spiders [ 10 , 11 ].…”

Section: Introductionmentioning

confidence: 99%

Isolation and Preliminary Characterization of Proteinaceous Toxins with Insecticidal and Antibacterial Activities from Black Widow Spider (L. tredecimguttatus) Eggs

Lei

et al. 2015

Toxins

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The eggs of black widow spider (L. tredecimguttatus) have been demonstrated to be rich in toxic proteinaceous components. The study on such active components is of theoretical and practical importance. In the present work, using a combination of multiple biochemical and biological strategies, we isolated and characterized the proteinaceous components from the aqueous extract of the black widow spider eggs. After gel filtration of the egg extract, the resulting main protein and peptide peaks were further fractionated by ion exchange chromatography and reversed-phase high performance liquid chromatography. Two proteinaceous components, named latroeggtoxin-III and latroeggtoxin-IV, respectively, were purified to homogeneity. Latroeggtoxin-III was demonstrated to have a molecular weight of about 36 kDa. Activity analysis indicated that latroeggtoxin-III exhibited neurotoxicity against cockroaches but had no obvious effect on mice, suggesting that it is an insect-specific toxin. Latroeggtoxin-IV, with a molecular weight of 3.6 kDa, was shown to be a broad-spectrum antibacterial peptide, showing inhibitory activity against all five species of bacteria tested, with the highest activity against Staphylococcus aureus. Finally, the implications of the proteinaceous toxins in egg protection and their potential applications were analyzed and discussed.

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Isolation and identification of a sodium channel-inhibiting protein from eggs of black widow spiders

Cited by 15 publications

References 20 publications

Transcriptome Analysis to Understand the Toxicity of Latrodectus tredecimguttatus Eggs

Transcriptome Analysis to Understand the Toxicity of Latrodectus tredecimguttatus Eggs

Physiological and biochemical characterization of egg extract of black widow spiders to uncover molecular basis of egg toxicity

Isolation and Preliminary Characterization of Proteinaceous Toxins with Insecticidal and Antibacterial Activities from Black Widow Spider (L. tredecimguttatus) Eggs

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