Photo-Cross-Linking of I Ks Demonstrates State-Dependent Interactions between KCNE1 and KCNQ1
| Autor: | Christopher I. Murray, Jodene Eldstrom, Maartje Westhoff, David Fedida |
|---|---|
| Rok vydání: | 2017 |
| Předmět: |
0301 basic medicine
chemistry.chemical_classification Membrane potential congenital hereditary and neonatal diseases and abnormalities endocrine system diseases urogenital system Stereochemistry Biophysics Cardiac action potential Gating Potassium channel Amino acid 03 medical and health sciences Transmembrane domain 030104 developmental biology 0302 clinical medicine Protein structure chemistry cardiovascular system Patch clamp 030217 neurology & neurosurgery |
| Zdroj: | Biophysical Journal. 113:415-425 |
| ISSN: | 0006-3495 |
| DOI: | 10.1016/j.bpj.2017.06.005 |
| Popis: | The slow delayed rectifier potassium current (IKs) is a key repolarizing current during the cardiac action potential. It consists of four KCNQ1 α-subunits and up to four KCNE1 β-subunits, which are thought to reside within external clefts of the channel. The interaction of KCNE1 with KCNQ1 dramatically delays opening of the channel but the mechanisms by which this occur are not yet fully understood. Here, we have used unnatural amino acid photo-cross-linking to investigate the dynamic interactions that occur between KCNQ1 and KCNE1 during activation gating. The unnatural amino acid p-Benzoylphenylalanine was successfully incorporated into two residues within the transmembrane domain of KCNE1: F56 and F57. UV-induced cross-linking suggested that F56Bpa interacts with KCNQ1 in the open state, whereas F57Bpa interacts predominantly in resting channel conformations. When UV was applied at progressively more depolarized preopen holding potentials, cross-linking of F57Bpa with KCNQ1 was slowed, which indicates that KCNE1 is displaced within the channel's cleft early during activation, or that conformational changes in KCNQ1 alter its interaction with KCNE1. In E1R/R4E KCNQ1, a mutant with constitutively activated voltage sensors, F56Bpa and F57Bpa KCNE1 were cross-linked in open and closed states, respectively, which suggests that their actions are mediated mainly by modulation of KCNQ1 pore function. |
| Databáze: | OpenAIRE |
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