The role of charged residues in independent glycine receptor folding domains for intermolecular interactions and ion channel function
| Autor: | Carmen Villmann, Georg Langlhofer |
|---|---|
| Rok vydání: | 2016 |
| Předmět: |
0301 basic medicine
Protein Folding Molecular Conformation Ionic bonding Biochemistry Ion Channels Ion 03 medical and health sciences Cellular and Molecular Neuroscience Receptors Glycine Humans Biotinylation Amino Acid Sequence Glycine receptor Ion channel Chemistry Intermolecular force Electrophysiological Phenomena Protein Structure Tertiary Folding (chemistry) Crystallography 030104 developmental biology HEK293 Cells Intramolecular force Biophysics Mutagenesis Site-Directed Ion Channel Gating Intracellular |
| Zdroj: | Journal of neurochemistry. 142(1) |
| ISSN: | 1471-4159 |
| Popis: | Glycine receptor (GlyR) truncations in the intracellular TM3-4 loop, documented in patients suffering from hyperekplexia and in the mouse mutant oscillator, lead to non-functionality of GlyRs. The missing part that contains the TM3-4 loop, TM4 and C-terminal sequences is essential for pentameric receptor arrangements. In vitro co-expressions of GlyRα1-truncated N-domains and C-domains were able to restore ion channel function. An ionic interaction between both domains was hypothesized as the underlying mechanism. Here, we analysed the proposed ionic interaction between GlyR N- and C-domains using C-terminal constructs with either positively or negatively charged N-termini. Charged residues at the N-terminus of the C-domain did interfere with receptor surface expression and ion channel function. In particular, presence of negatively charged residues at the N-terminus led to significantly decreased ion channel function. Presence of positive charges resulted in reduced maximal currents possibly as a result of repulsion of both domains. If the C-domain was tagged by a myc-epitope, low maximal current amplitudes were detected. Intrinsic charges of the myc-epitope and charged N-terminal ends of the C-domain most probably induce intramolecular interactions. These interactions might hinder the close proximity of C-domains and N-domains, which is a prerequisite for functional ion channel configurations. The remaining basic subdomains close to TM3 and 4 were sufficient for domain complementation and functional ion channel formation. Thus, these basic subdomains forming α-helical elements or an intracellular portal represent attractants for incoming negatively charged chloride ions and interact with the phospholipids thereby stabilizing the GlyR in a conformation that allows ion channel opening. |
| Databáze: | OpenAIRE |
| Externí odkaz: |
|
Plný text