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  Domain 2 of Drosophila Para voltage-gated sodium channel confers insect properties to a rat brain channel

Shichor, I., Zlotkin, E., Ilan, N., Chikashvili, D., Stühmer, W., Gordon, D., et al. (2002). Domain 2 of Drosophila Para voltage-gated sodium channel confers insect properties to a rat brain channel. The Journal of Neuroscience: the Official Journal of the Society for Neuroscience, 22(11), 4364-4371.

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Shichor, I., Author
Zlotkin, E., Author
Ilan, N., Author
Chikashvili, D., Author
Stühmer, Walter1, Author              
Gordon, D., Author
Lotan, I., Author
Affiliations:
1Molecular biology of neuronal signals, Max Planck Institute of Experimental Medicine, Max Planck Society, ou_2173656              

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Free keywords: Na channel; insect selectivity; Xenopus oocytes; scorpion toxin; gating; Drosophila Para
 Abstract: The ability of the excitatory anti-insect-selective scorpion toxin AahIT (Androctonus australis hector) to exclusively bind to and modify the insect voltage-gated sodium channel (NaCh) makes it a unique tool to unravel the structural differences between mammalian and insect channels, a prerequisite in the design of selective pesticides. To localize the insect NaCh domain that binds AahIT, we constructed a chimeric channel composed of rat brain NaCh alpha-subunit (rBIIA) in which domain-2 (D2) was replaced by that of Drosophila Para (paralytic temperature-sensitive). The choice of D2 was dictated by the similarity between AahIT and scorpion beta- toxins pertaining to both their binding and action and the essential role of D2 in the beta-toxins binding site on mammalian channels. Expression of the chimera rBIIA-ParaD2 in Xenopus oocytes gave rise to voltage-gated and TTX-sensitive NaChs that, like rBIIA, were sensitive to scorpion alpha-toxins and regulated by the auxiliary subunit beta(1) but not by the insect TipE. Notably, like Drosophila Para/TipE, but unlike rBIIA/beta(1), the chimera gained sensitivity to AahIT, indicating that the phyletic selectivity of AahIT is conferred by the insect NaCh D2. Furthermore, the chimera acquired additional insect channel properties; its activation was shifted to more positive potentials, and the effect of alpha- toxins was potentiated. Our results highlight the key role of D2 in the selective recognition of anti-insect excitatory toxins and in the modulation of NaCh gating. We also provide a methodological approach to the study of ion channels that are difficult to express in model expression systems.

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Language(s): eng - English
 Dates: 2002-06-01
 Publication Status: Published in print
 Pages: -
 Publishing info: -
 Table of Contents: -
 Rev. Type: Peer
 Identifiers: eDoc: 17464
ISI: 000175900700016
 Degree: -

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Title: The Journal of Neuroscience : the Official Journal of the Society for Neuroscience
  Other : J. Neurosci.
Source Genre: Journal
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Publ. Info: Baltimore, MD : The Society
Pages: - Volume / Issue: 22 (11) Sequence Number: - Start / End Page: 4364 - 4371 Identifier: ISSN: 0270-6474
CoNE: https://pure.mpg.de/cone/journals/resource/954925502187_1