title: Ion channels of N-terminally linked alamethicin dimers: enhancement of cation-selectivity by substitution of Glu for Gln at position 7.
abstract: Alamethicin forms voltage-gated ion channels that have moderate cation-selectivity. The enhancement of the cation-selectivity by introducing negatively charged residues at positions 7 and 18 has been studied using the tethered homodimers of alamethicin with Q7 and E18 (di-alm-Q7E18) and its analog with E7 and Q18 (di-alm-E7Q18). In the dimeric peptides, monomer peptides are linked at the N-termini by a disulfide bond. Both the peptides formed long lasting ion channels at cis-positive voltages when added to the cis-side membrane. Their long open duration enabled us to obtain current-voltage (I-V(m)) relations and reversal potentials at the single-channel level by applying a voltage ramp during the channel opening. The reversal potentials measured in asymmetric KCl solutions indicated that ionized E7 provided strong cation-selectivity, whereas ionized E18 little influenced the charge selectivity. This was also the case for the macroscopic charge selectivity determined from the reversal potentials obtained by the macroscopic I-V(m) measurements. The results are accounted for by stronger electrostatic interactions between permeant ions and negatively charged residues at the narrowest part of the pore than at the pore mouth.
authors.affiliation: Graduate School of Science, Osaka University, Toyonaka 560-0043, Japan.
authors.forename: Takashi
authors.initials: T
authors.lastname: Okazaki
authors.forename: Yasuo
authors.initials: Y
authors.lastname: Nagaoka
authors.forename: Koji
authors.initials: K
authors.lastname: Asami
mesh_terms: D000408:Alamethicin / Q000737:chemistry*; D019943:Amino Acid Substitution; D002412:Cations; D019281:Dimerization; D004553:Electric Conductivity; D004563:Electrochemistry / Q000379:methods*; D018698:Glutamic Acid / Q000737:chemistry*; D005973:Glutamine / Q000737:chemistry*; D015640:Ion Channel Gating; D007473:Ion Channels / Q000737:chemistry*; D012680:Sensitivity and Specificity; D013329:Structure-Activity Relationship
chemical_list: D002412:Cations; D007473:Ion Channels; D005973:Glutamine; D000408:Alamethicin; D018698:Glutamic Acid