 |
 Previous Article | Next Article 
The Journal of Neuroscience, June 1, 2002, 22(11):4364-4371
Domain 2 of Drosophila Para Voltage-Gated Sodium
Channel Confers Insect Properties to a Rat Brain Channel
Iris
Shichor1, 3,
Eliahu
Zlotkin3,
Nitza
Ilan1, 2,
Dodo
Chikashvili1,
Walter
Stuhmer4,
Dalia
Gordon2, and
Ilana
Lotan1
Departments of 1 Physiology and Pharmacology,
Sackler School of Medicine and 2 Plant Sciences, George S. Wise Faculty of Life Sciences, Tel-Aviv University, 69978 Ramat-Aviv,
Israel, 3 Department of Cell and Animal Biology, Institute
of Life Sciences, The Hebrew University, 91904 Jerusalem, Israel, and
4 Max-Planck-Institut fur Experimentelle Medizin, D-37075
Gottingen, Germany
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
 -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  -toxins pertaining to both
their binding and action and the essential role of D2 in the -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 -toxins and regulated by the auxiliary subunit
1 but not by the insect TipE. Notably, like Drosophila Para/TipE, but unlike
rBIIA/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 -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.
Key words:
Na channel; insect selectivity; Xenopus
oocytes; scorpion toxin; gating; Drosophila Para
Copyright © 2002 Society for Neuroscience 0270-6474/02/22114364-08$05.00/0
This article has been cited by other articles:
|
|
|
|
 
L. Cohen, Y. Moran, A. Sharon, D. Segal, D. Gordon, and M. Gurevitz
Drosomycin, an Innate Immunity Peptide of Drosophila melanogaster, Interacts with the Fly Voltage-gated Sodium Channel
J. Biol. Chem.,
August 28, 2009;
284(35):
23558 - 23563.
[Abstract]
[Full Text]
[PDF]
|
|
|
|
|
|
|
R. Kahn, I. Karbat, N. Ilan, L. Cohen, S. Sokolov, W. A. Catterall, D. Gordon, and M. Gurevitz
Molecular Requirements for Recognition of Brain Voltage-gated Sodium Channels by Scorpion {alpha}-Toxins
J. Biol. Chem.,
July 31, 2009;
284(31):
20684 - 20691.
[Abstract]
[Full Text]
[PDF]
|
|
|
|
|
|
|
L. Cohen, N. Lipstein, I. Karbat, N. Ilan, N. Gilles, R. Kahn, D. Gordon, and M. Gurevitz
Miniaturization of Scorpion {beta}-Toxins Uncovers a Putative Ancestral Surface of Interaction with Voltage-gated Sodium Channels
J. Biol. Chem.,
May 30, 2008;
283(22):
15169 - 15176.
[Abstract]
[Full Text]
[PDF]
|
|
|
|
|
|
|
L. Cohen, Y. Troub, M. Turkov, N. Gilles, N. Ilan, M. Benveniste, D. Gordon, and M. Gurevitz
Mammalian Skeletal Muscle Voltage-Gated Sodium Channels Are Affected by Scorpion Depressant "Insect-Selective" Toxins when Preconditioned
Mol. Pharmacol.,
November 1, 2007;
72(5):
1220 - 1227.
[Abstract]
[Full Text]
[PDF]
|
|
|
|
|
|
|
L. Cohen, N. Ilan, M. Gur, W. Stuhmer, D. Gordon, and M. Gurevitz
Design of a Specific Activator for Skeletal Muscle Sodium Channels Uncovers Channel Architecture
J. Biol. Chem.,
October 5, 2007;
282(40):
29424 - 29430.
[Abstract]
[Full Text]
[PDF]
|
|
|
|
|
|
|
G. Corzo, J. K. Sabo, F. Bosmans, B. Billen, E. Villegas, J. Tytgat, and R. S. Norton
Solution Structure and Alanine Scan of a Spider Toxin That Affects the Activation of Mammalian Voltage-gated Sodium Channels
J. Biol. Chem.,
February 16, 2007;
282(7):
4643 - 4652.
[Abstract]
[Full Text]
[PDF]
|
|
|
|
|
|
|
L. Cohen, N. Lipstein, and D. Gordon
Allosteric interactions between scorpion toxin receptor sites on voltage-gated Na channels imply a novel role for weakly active components in arthropod venom
FASEB J,
September 1, 2006;
20(11):
1933 - 1935.
[Abstract]
[Full Text]
[PDF]
|
|
|
|
|
|
|
L. Cohen, N. Gilles, I. Karbat, N. Ilan, D. Gordon, and M. Gurevitz
Direct Evidence That Receptor Site-4 of Sodium Channel Gating Modifiers Is Not Dipped in the Phospholipid Bilayer of Neuronal Membranes
J. Biol. Chem.,
July 28, 2006;
281(30):
20673 - 20679.
[Abstract]
[Full Text]
[PDF]
|
|
|
|
|
|
|
E. Leipold, A. Hansel, A. Borges, and S. H. Heinemann
Subtype Specificity of Scorpion beta-Toxin Tz1 Interaction with Voltage-Gated Sodium Channels Is Determined by the Pore Loop of Domain 3
Mol. Pharmacol.,
July 1, 2006;
70(1):
340 - 347.
[Abstract]
[Full Text]
[PDF]
|
|
|
|
|
|
|
K. IZHAR, C. LIOR, G. NICOLAS, G. DALIA, and G. MICHAEL
Conversion of a scorpion toxin agonist into an antagonist highlights an acidic residue involved in voltage sensor trapping during activation of neuronal Na+ channels
FASEB J,
April 1, 2004;
18(6):
683 - 689.
[Abstract]
[Full Text]
[PDF]
|
|
|
|
|
|
|
L. Cohen, I. Karbat, N. Gilles, O. Froy, G. Corzo, R. Angelovici, D. Gordon, and M. Gurevitz
Dissection of the Functional Surface of an Anti-insect Excitatory Toxin Illuminates a Putative "Hot Spot" Common to All Scorpion {beta}-Toxins Affecting Na+ Channels
J. Biol. Chem.,
February 27, 2004;
279(9):
8206 - 8211.
[Abstract]
[Full Text]
[PDF]
|
|
|
|
|
