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Mutations in the pore-forming subunit of the ATP-sensitive K(+) (K(ATP)) channel Kir6.2 cause neonatal diabetes. Understanding the molecular mechanism of action of these mutations has provided valuable insight into the relationship between the structure and function of the K(ATP) channel. When Kir6.2 containing a mutation (F333I) in the putative ATP-binding site is coexpressed with the cardiac type of regulatory K(ATP) channel subunit, SUR2A, the channel sensitivity to ATP inhibition is reduced and the intrinsic open probability (P(o)) is increased. However, the extent of macroscopic current activation by MgADP was unaffected. Here we examine rundown and MgADP activation of wild-type and Kir6.2-F333I/SUR2A channels using single-channel recording, noise analysis and spectral analysis. We also compare the effect of mutating the adjacent residue, G334, on rundown and MgADP activation. All three approaches indicated that rundown of Kir6.2-F333I/SUR2A channels is due to a reduction in the number of active channels in the patch and that MgADP reactivation involves recruitment of inactive channels. In contrast, rundown and MgADP reactivation of wild-type and Kir6.2-G334D/SUR2A channels, and of Kir6.2-F333I/SUR1 channels, involve a gradual change in P(o). Our results suggest that F333 in Kir6.2 interacts functionally with SUR2A to modulate channel rundown and MgADP activation. This interaction is fairly specific as it is not disturbed when the adjacent residue (G334) is mutated. It is also not a consequence of the enhanced P(o) of Kir6.2-F333I/SUR2A channels, as it is not found for other mutant channels with high P(o) (Kir6.2-I296L/SUR2A).

Original publication

DOI

10.1113/jphysiol.2007.143149

Type

Journal article

Journal

J Physiol

Publication Date

01/11/2007

Volume

584

Pages

743 - 753

Keywords

ATP-Binding Cassette Transporters, Adenosine Diphosphate, Adenosine Triphosphate, Animals, Binding Sites, Electrophysiology, Humans, KATP Channels, Molecular Biology, Mutation, Oocytes, Potassium Channels, Inwardly Rectifying, Protein Binding, Rats, Receptors, Drug, Sulfonylurea Receptors, Xenopus laevis