Journal
INTERNATIONAL JOURNAL OF BIOLOGICAL MACROMOLECULES
Volume 65, Issue -, Pages 115-120Publisher
ELSEVIER
DOI: 10.1016/j.ijbiomac.2014.01.004
Keywords
Black widow spider; Egg; Toxin; Protein; Isolation; Characterization
Funding
- National Natural Science Foundation of China [31070700, 31271135]
- National Basic Research Program or 973 Program of China [2010CB529800]
- cooperative Innovation Center of Engineering and New Products for Developmental Biology of Hunan Province
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The eggs of black widow spider (L. tredecimguttatus) have been demonstrated to be rich in biologically active components that exhibit great research value and application foreground. In the present study, a protein toxin, named Latroeggtoxin-II, was isolated from the eggs using the combination of gel filtration, ion exchange chromatography and reversed-phase high performance liquid chromatography. Electraspray mass spectrometric analysis indicated that the molecular weight of the protein was 28.69 kDa, and Edman degradation revealed that its N-terminal sequence was ESIQT STYVP NTPNQKFDYEVGKDY-. After being abdominally injected into mice and P. americana, the protein could make the animals especially P. americana display a series of poisoning symptoms. Electrophysiological experiments demonstrated that the protein could selectively inhibit tetrodotoxin-resistant Na+ channel currents in rat dorsal root ganglion neurons, without significant effect on the tetrodotoxin-sensitive Na+ channel currents. Using multiple proteomic strategies, the purified protein was shown to have only a few similarities to the existing proteins in the databases, suggesting that it was a novel protein isolated from the eggs of black widow spiders. (C) 2014 Published by Elsevier B.V.
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