Beta-scorpion toxin effects suggest electrostatic interactions in domain II of voltage-dependent sodium channels.
Beta-scorpion toxins specifically modulate the voltage dependence of sodium channel activation by acting through a voltage-sensor trapping model. We used mutagenesis, functional analysis and the action of beta-toxin as tools to investigate the existence and role in channel activation of molecular interactions between the charged residues of the S2, S3 and S4 segments in domain II of sodium channels. Mutating to arginine the acidic residues of the S2 and S3 transmembrane segments in domain II, or making charge-reversal mutation of the two outermost gating charges of the IIS4 voltage sensor, shifts the voltage dependence of channel activation to more positive potentials and enhances the effect of beta-scorpion toxin. Thus, mutations of acidic residues in IIS2 and IIS3 segments are able to promote voltage-sensor trapping in a way that is similar to the mutations of the arginines in the IIS4 segment. In order to disclose the network of interactions among acidic and basic residues we performed functional analysis of charge-inversion double mutants: our data suggest that the first arginine of the voltage sensor S4 in domain II (R850) interacts specifically with E805, D814 and E821 in the S2 and S3 segments, whereas the second arginine (R853) only interacts with D827 in the S3 segment. Our results suggest that the S2, S3 and S4 segments in domain II form a voltage-sensing structure, and that molecular interactions between the charged residues of this structure modulate the availability of the IIS4 voltage sensor for trapping by beta-toxins. They also provide unique insights into the molecular events that occur during channel activation, as well as into the structure of the channel.
Mantegazza
Massimo
M
Dipartimento di Neurofisiopatologia, Istituto Neurologico Besta, Milano, Italy.
Cestèle
Sandrine
S
D024342:Amino Acids, Acidic / Q000737:chemistry; D000818:Animals; D001120:Arginine / Q000737:chemistry; D001921:Brain; D002460:Cell Line; D006801:Humans; D015640:Ion Channel Gating / Q000187:drug effects; D007668:Kidney; D008564:Membrane Potentials / Q000187:drug effects; D009154:Mutation / Q000187:drug effects; D062551:NAV1.2 Voltage-Gated Sodium Channel; D009419:Nerve Tissue Proteins / Q000187:drug effects* / Q000235:genetics / Q000378:metabolism; D017434:Protein Structure, Tertiary; D051381:Rats; D012604:Scorpion Venoms / Q000494:pharmacology*; D015222:Sodium Channels / Q000187:drug effects* / Q000235:genetics / Q000378:metabolism; D055672:Static Electricity; D014162:Transfection
D024342:Amino Acids, Acidic; C115646:CSS IV toxin; D062551:NAV1.2 Voltage-Gated Sodium Channel; D009419:Nerve Tissue Proteins; C568251:SCN2A protein, human; C568253:Scn2A protein, rat; D012604:Scorpion Venoms; D015222:Sodium Channels; D001120:Arginine