Plant Kin and Kout channels: Approaching the trait of opposite rectification by analyzing more than 250 KAT1-SKOR chimeras
| Autor: | Klaas Wulfetange, Hervé Sentenac, Ingo Dreyer, Fabien Porée, Armando Carpaneto, Adam Bertl, Jean-Baptiste Thibaud, Anja Roller, Alessia Naso, Gabriel Natura |
|---|---|
| Jazyk: | angličtina |
| Rok vydání: | 2005 |
| Předmět: |
Potassium Channels
Molecular Sequence Data Biophysics Yeast complementation Rectification Biology Biochemistry Membrane Potentials 03 medical and health sciences Chimera (genetics) Structure-Activity Relationship Chimera Potassium channel Site-directed mutagenesis Amino Acid Sequence Amino Acid Substitution Arabidopsis Proteins Ion Channel Gating Potassium Potassium Channels Inwardly Rectifying Protein Isoforms Recombinant Proteins Shaker Superfamily of Potassium Channels Molecular Biology 030304 developmental biology Genetics 0303 health sciences 030302 biochemistry & molecular biology Depolarization Cell Biology Yeast strain Triple mutant Cell biology Inwardly Rectifying Communication channel |
| Popis: | Members of the Shaker-like plant K(+) channel family share a common structure, but are highly diverse in their function: they behave as either hyperpolarization-activated inward-rectifying (K(in)) channels, or leak-like (K(weak)) channels, or depolarization-activated outward-rectifying (K(out)) channels. Here we created 256 chimeras between the K(in) channel KAT1 and the K(out) channel SKOR. The chimeras were screened in a potassium-uptake deficient yeast strain to identify those, which mediate potassium inward currents, i.e., which are functionally equivalent to KAT1. This strategy allowed us to identify three chimeras which differ from KAT1 in three parts of the polypeptide: the cytosolic N-terminus, the cytosolic C-terminus, and the putative voltage-sensor S4. Additionally, mutations in the K(out) channel SKOR were generated in order to localize molecular entities underlying its depolarization activation. The triple mutant SKOR-D312N-M313L-I314G, carrying amino-acid changes in the S6 segment, was identified as a channel which did not display any rectification in the tested voltage-range. |
| Databáze: | OpenAIRE |
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