Involvement of different S4 parts in the voltage dependency of Na channel gating.
Three synthetic peptides corresponding to parts of S4 of the first repeat of eel electroplax sodium channel were synthesized. The basic peptide was C1+ which corresponds to amino acids 210-223 (eel channel numbering) and two subfractions: an external fraction, C1+ex (amino acid 210-217); and an internal part, C1+in (amino acid 218-221). Peptide C1+ includes four of the charged amino acids of this domain; peptide C1+ex includes three of the charged amino acids and is closer to the external membrane surface (according to channel models) than peptide C1+in which includes the fourth charged amino acid alone. Antibodies generated in rabbits against these peptides were shown to be site specific. Using the whole-cell patch-clamp technique, we found that in rat dorsal root ganglion (DRG) cells, the antibodies against C1+in but not against C1+ex had an effect on the gating parameters. They shifted the Na-channel inactivation curve towards hyperpolarization and decreased the slope of the Na-channel activation curve. These results demonstrate that during the conformational changes associated with channel gating, the fourth charged amino acid of S4 must be accessible to antibodies given to the external solution. Furthermore, they indicate a specific involvement of S4 in the voltage dependency of the gating processes.
Kra-Oz
Z
Z
Department of Physiology and Biophysics, Bruce Rappaport Faculty of Medicine, Technion-Israel Institute of Technology, Haifa.
Spira
G
G
Palti
Y
Y
Meiri
H
H
D000595:Amino Acid Sequence; D000818:Animals; D000906:Antibodies / Q000378:metabolism; D002478:Cells, Cultured; D004594:Electrophysiology; D004797:Enzyme-Linked Immunosorbent Assay; D005727:Ganglia, Spinal / Q000166:cytology / Q000502:physiology / Q000648:ultrastructure; D015640:Ion Channel Gating / Q000502:physiology*; D008969:Molecular Sequence Data; D051381:Rats; D015222:Sodium Channels / Q000737:chemistry / Q000378:metabolism / Q000502:physiology*
D000906:Antibodies; D015222:Sodium Channels