Abstract
1. Two mutants of the sodium channel II have been expressed inXenopus oocytes and have been investigated using the patch-clamp technique. In mutant E387Q the glutamic acid at position 387 has been replaced by glutamine, and in mutant D384N the aspartic acid at position 384 has been replaced by asparagine.2. Mutant E387Q, previously shown to be resistant to block by tetrodotoxin (Noda et al. 1989), has a single-channel conductance of 4 pS, that can be easily measured only using noise analysis. At variance with the wild-type, the openchannel current-voltage relationship of mutant E387Q is linear over a wide voltage range even under asymmetrical ionic conditions.3. Mutant D384N has a very low permeability for any of the following ions: Cl−, Na+, K+, Li+, Rb+, Ca2+, Mg2+, NH4 + , TMA+, TEA+. However, asymmetric charge movements similar to the gating currents of the Na+-selective wild-type are still observed.4. These results suggest that residues E387 and D384 interact directly with the pathway of the ions permeating the open channel.
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Abbreviations
- TTX:
-
tetrodotoxin; Na+, sodium; K+, potassium;
- NFR:
-
normal frog Ringer
- HEPES:
-
N-2-hydroxylethyl piperazine-N′-2-ethanesulfonic acid
- EGTA:
-
ethyleneglycol-bis(β-amino-ethyl ether) N,N,N',N'-tetra acetic acid
- TEA:
-
tetraethylammonium
- TMA:
-
tetramethylammonium;I g , gating current; γ, single-channel conductance
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Pusch, M., Noda, M., Stühmer, W. et al. Single point mutations of the sodium channel drastically reduce the pore permeability without preventing its gating. Eur Biophys J 20, 127–133 (1991). https://doi.org/10.1007/BF01561134
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DOI: https://doi.org/10.1007/BF01561134