Mutational analysis of state-dependent contacts in the pore module of eukaryotic sodium channels.
Voltage-gated sodium channels have residues that change or may change contacts upon gating. Contributions of individual contacts in stability of different states are incompletely understood. Pore-lining inner helices contain exceptionally conserved asparagines in positions i20. Here we explored how mutations in positions i20 and i29 affect electrophysiological properties of insect sodium channels. In repeat interfaces I/IV, III/II and IV/III, alanine substitutions caused positive activation shifts in positions i20 and i29, negative shifts of slow inactivation in positions i20 and positive shifts of slow inactivation in positions i29. The results support the hypothesis on open state inter-repeat H-bonding of residues i20 and i29. The shift magnitudes vary between interfaces, reflecting structural asymmetry of the channels. Mutations in positions i20 of repeats III and IV caused much longer recovery delay from the slow and fast inactivation than other mutations. In repeat IV, alanine substitution of tyrosine i30 rescued positive activation shift of mutation in position i29. Our data suggest that polar residues in positions i29 are involved in stabilization of both the open and slow-inactivated states. Transition between the states may involve switching of H-bonding partners of residues i29 from the conserved asparagines to currently unknown residues.
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Related Subject Headings
- Sodium Channels
- Sequence Homology, Amino Acid
- Mutation
- Mutagenesis, Site-Directed
- Ion Channel Gating
- Insecta
- Biochemistry & Molecular Biology
- Animals
- Amino Acid Sequence
- 0601 Biochemistry and Cell Biology
Citation
Published In
DOI
EISSN
ISSN
Publication Date
Volume
Start / End Page
Related Subject Headings
- Sodium Channels
- Sequence Homology, Amino Acid
- Mutation
- Mutagenesis, Site-Directed
- Ion Channel Gating
- Insecta
- Biochemistry & Molecular Biology
- Animals
- Amino Acid Sequence
- 0601 Biochemistry and Cell Biology