Substitution of a hydrophobic residue alters the conformational stability of Shaker K+ channels during gating and assembly.
A leucine residue at position 370 (L370) in 29-4 Shaker K+ channels resides within two overlapping sequence motifs conserved among most voltage-gated channels: the S4 segment and a leucine heptad repeat. Here we investigate the effects observed upon substitution of L370 with many other uncharged amino acid residues. We find that smaller or more hydrophilic residues produce greater alterations in both activation and inactivation gating than does substitution with other large hydrophobic residues. In addition, subunits containing less conservative substitutions at position 370 are restricted in their assembly with wild-type subunits and are unlikely to form homomultimeric channel complexes. Consistent with the idea that L370 influences the tertiary structure of these channels, the results indicate that L370 undergoes specific hydrophobic interactions during the conformational transitions of gating; similar interactions may take place during the folding, insertion, or assembly of Shaker K+ channel subunits.
McCormack
K
K
Department of Cellular and Molecular Physiology, Yale University School of Medicine, New Haven, Connecticut 06510.
Lin
L
L
Sigworth
F J
FJ
D000595:Amino Acid Sequence; D000818:Animals; D001483:Base Sequence; D055592:Biophysical Phenomena; D001703:Biophysics; D018076:DNA, Complementary / Q000235:genetics; D004330:Drosophila; D004355:Drug Stability; D005260:Female; D015640:Ion Channel Gating; D008564:Membrane Potentials; D008969:Molecular Sequence Data; D016297:Mutagenesis, Site-Directed; D009865:Oocytes / Q000378:metabolism; D010455:Peptides / Q000737:chemistry / Q000235:genetics / Q000378:metabolism*; D015221:Potassium Channels / Q000737:chemistry / Q000235:genetics / Q000378:metabolism*; D011487:Protein Conformation; D051637:Shaker Superfamily of Potassium Channels; D013816:Thermodynamics; D014982:Xenopus laevis
D018076:DNA, Complementary; D010455:Peptides; D015221:Potassium Channels; C076413:Shaker B potassium channel polypeptide; D051637:Shaker Superfamily of Potassium Channels