Structural study of the sodium channel inactivation gate peptide including an isoleucine-phenylalanine-methionine motif and its analogous peptide (phenylalanine/glutamine) in trifluoroethanol solutions and SDS micelles.
In order to gain insight into the gating mechanisms of Na+ channels, in particular their inactivation mechanisms, we studied the structures of the Na+ channel inactivation gate related peptide which includes the IFM (Ile-Phe-Met) motif (Ac-KKKFGGQDIFMTEEQKK-NH2; K1480-K1496 in rat brain type-IIA Na+ channels, MP-3A) and its F/Q(Gln) substituted one (MP-4A) in trifluoroethanol (TFE) solutions and sodium dodecyl sulfate (SDS) micelles using circular dichroism (CD) and 1H-NMR spectroscopies. Based on observed nuclear Overhauser effect constraints, three-dimensional structures of MP-3A and MP-4A were determined using simulated annealing molecular dynamics/ energy minimization calculations. In TFE solutions, no appreciable differences in the structure were observed using either CD or NMR spectra. In SDS micelles, however, the two peptides exhibited definitely different structures from each other. It was found that in MP-3A, residues 11488 and T1491 were spatially proximate with each other owing to hydrogen bonding between the amide proton of 11488 and the hydroxyl oxygen atom of T1491, whereas in MP-4A, F/Q substitution separated them owing to conformational changes. The solvent-accessible surfaces calculated for the structures of MP-3A and MP-4A showed that the former has a smoother interaction surface to the hydrophobic docking site than the latter. In conclusion, the conformational changes, as well as decreased hydrophobicity around the IFM motif owing to the F/Q mutation, may be one reason why F1489Q mutated channels cannot inactivate almost completely.
Kuroda
Y
Y
Graduate School of Pharmaceutical Sciences, Kyoto University, Japan. yokuroda@pharm.kyoto-u.ac.jp
Miyamoto
K
K
Matsumoto
M
M
Maeda
Y
Y
Kanaori
K
K
Otaka
A
A
Fujii
N
N
Nakagawa
T
T
D020816:Amino Acid Motifs; D002942:Circular Dichroism; D015640:Ion Channel Gating; D007532:Isoleucine / Q000737:chemistry*; D008715:Methionine / Q000737:chemistry*; D019906:Nuclear Magnetic Resonance, Biomolecular / Q000379:methods; D009842:Oligopeptides / Q000737:chemistry* / Q000494:pharmacology; D010446:Peptide Fragments / Q000737:chemistry*; D010649:Phenylalanine / Q000737:chemistry*; D026941:Sodium Channel Blockers; D012967:Sodium Dodecyl Sulfate / Q000737:chemistry*; D014270:Trifluoroethanol / Q000737:chemistry*
D009842:Oligopeptides; D010446:Peptide Fragments; D026941:Sodium Channel Blockers; D007532:Isoleucine; D012967:Sodium Dodecyl Sulfate; D010649:Phenylalanine; D014270:Trifluoroethanol; D008715:Methionine