Role of the S4 segment in a voltage-dependent calcium-sensitive potassium (hSlo) channel

Diaz L.; Amigo, J; Alvarez, O; Latorre R.; Stefani E.; Meera, P; Toro, L.

Keywords: sequence, acid, membrane, animals, ion, protein, cell, gene, conductance, channel, calcium, mutation, laevis, channels, virus, site, humans, potassium, mutagenesis, domain, level, xenopus, rearrangement, molecular, data, article, hydrophobicity, gating, point, neutralization, amino, priority, nonhuman, journal, potential, directed


We investigated the role of individual charged residues of the S4 region of a MaxiK channel (hSlo) in channel gating. We measured macroscopic currents induced by wild type (WT) and point mutants of hSlo in inside-out membrane patches of Xenopus laevis oocytes. Of all the residues tested, only neutralizations of Arg-210 and Arg-213 were associated with a reduction in the number of gating charges as determined using the limiting slope method. Channel activation in WT and mutant channels was interpreted using an allosteric model. Mutations R207Q, R207E, and R210N facilitated channel opening in the absence of Ca 2+; however, this facilitation was not observed in the channels Ca 2+-bound state. Mutation R213Q behaved similarly to the WT channel in the absence of Ca 2+, but Ca 2+ was unable to stabilize the open state to the same extent as it does in the WT. Mutations R207Q, R207E, R210N, and R213Q reduced the coupling between Ca 2+ binding and channel opening when compared with the WT. Mutations L204R, L204H, Q216R, E219Q, and E219K in the S4 domain showed a similar phenotype to the WT channel. We conclude that the S4 region in the hSlo channel is part of the voltage sensor and that only two charged amino acid residues in this region (Arg-210 and Arg-213) contribute to the gating valence of the channel.

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Volumen: 273
Número: 49
Editorial: Elsevier
Fecha de publicación: 1998
Página de inicio: 32430
Página final: 32436