Ion channel formation by synthetic transmembrane segments of the inhibitory glycine receptor--a model study.
The inhibitory glycine receptor (GlyR) of rat spinal cord contains an intrinsic transmembrane channel mediating agonist-gated anion flux. Here, synthetic peptides modelled after the predicted transmembrane domains M2 and M4 of its ligand-binding subunit were incorporated into lipid vesicle membranes and black lipid bilayers to analyze their channel forming capabilities. Both types of peptides prohibited the establishment of, or dissipated, preexisting transmembrane potentials in the vesicle system. Incorporation of peptide M2 into the black lipid bilayer elicited randomly gated single channel events with various conductance states and life-times. Peptide M4 increased the conductance of the bilayer without producing single channels. Exchange of the terminal arginine residues of peptide M2 by glutamate resulted in a significant shift towards cation selectivity of the respective channels as compared to peptide M2. In conclusion, the peptide channels observed differed significantly from native GlyR in both conductivity and ion-selectivity indicating that individual synthetic transmembrane segments are not sufficient to mimic a channel protein composed of subunits with multiple transmembrane segments.
Langosch
D
D
ZMBH, Universität Heidelberg, F.R.G.
Hartung
K
K
Grell
E
E
Bamberg
E
E
Betz
H
H
D000595:Amino Acid Sequence; D002942:Circular Dichroism; D004553:Electric Conductivity; D066298:In Vitro Techniques; D015640:Ion Channel Gating; D007473:Ion Channels / Q000737:chemistry*; D008051:Lipid Bilayers; D008564:Membrane Potentials; D008565:Membrane Proteins / Q000737:chemistry; D008969:Molecular Sequence Data; D010446:Peptide Fragments; D011487:Protein Conformation; D018009:Receptors, Glycine; D017981:Receptors, Neurotransmitter / Q000737:chemistry*; D012995:Solubility
D007473:Ion Channels; D008051:Lipid Bilayers; D008565:Membrane Proteins; D010446:Peptide Fragments; D018009:Receptors, Glycine; D017981:Receptors, Neurotransmitter