BjalphaIT: a novel scorpion alpha-toxin selective for insects--unique pharmacological tool.
Long-chain neurotoxins derived from the venom of the Buthidae scorpions, which affect voltage-gated sodium channels (VGSCs) can be subdivided according to their toxicity to insects into insect-selective excitatory and depressant toxins (beta-toxins) and the alpha-like toxins which affect both mammals and insects. In the present study by the aid of reverse-phase HPLC column chromatography, RT-PCR, cloning and various toxicity assays, a new insect selective toxin designated as BjalphaIT was isolated from the venom of the Judean Black Scorpion (Buthotus judaicus), and its full primary sequence was determined: MNYLVVICFALLLMTVVESGRDAYIADNLNCAYTCGSNSYCNTECTKNGAVSGYCQWLGKYGNACWCINLPDKVPIRIPGACR (leader sequence is underlined). Despite its lack of toxicity to mammals and potent toxicity to insects, BjalphaIT reveals an amino acid sequence and an inferred spatial arrangement that is characteristic of the well-known scorpion alpha-toxins highly toxic to mammals. BjalphaITs sharp distinction between insects and mammals was also revealed by its effect on sodium conductance of two cloned neuronal VGSCs heterloguously expressed in Xenopus laevis oocytes and assayed with the two-electrode voltage-clamp technique. BjalphaIT completely inhibits the inactivation process of the insect para/tipE VGSC at a concentration of 100 nM, in contrast to the rat brain Na(v)1.2/beta1 which is resistant to the toxin. The above categorical distinction between mammal and insect VGSCs exhibited by BjalphaIT enables its employment in the clarification of the molecular basis of the animal group specificity of scorpion venom derived neurotoxic polypeptides and voltage-gated sodium channels.
Arnon
Tal
T
Department of Cell and Animal biology, Institute of Life Sciences, The Hebrew University of Jerusalem, Jerusalem 91904, Israel.
Potikha
Tamara
T
Sher
Daniel
D
Elazar
Menashe
M
Mao
Wenfu
W
Tal
Tzachy
T
Bosmans
Frank
F
Tytgat
Jan
J
Ben-Arie
Nissim
N
Zlotkin
Eliahu
E
D000595:Amino Acid Sequence; D000818:Animals; D007313:Insecta; D008969:Molecular Sequence Data; D017434:Protein Structure, Tertiary; D012604:Scorpion Venoms / Q000737:chemistry* / Q000235:genetics / Q000302:isolation & purification*; D012605:Scorpions / Q000737:chemistry* / Q000235:genetics; D013045:Species Specificity
D012604:Scorpion Venoms; C500707:alphaIT toxin, Buthotus judaicus