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Data from: Activation mechanism of ATP-sensitive K+ channels explored with real-time nucleotide binding

Citation

Puljung, Michael; Vedovato, Natascia; Usher, Samuel; Ashcroft, Frances (2019), Data from: Activation mechanism of ATP-sensitive K+ channels explored with real-time nucleotide binding, Dryad, Dataset, https://doi.org/10.5061/dryad.6mh0sv3

Abstract

The response of ATP-sensitive K+ channels (KATP) to cellular metabolism is coordinated by three classes of nucleotide binding site (NBS). We used a novel approach involving labeling of intact channels in a native, membrane environment with a non-canonical fluorescent amino acid and measurement (using FRET with fluorescent nucleotides) of steady-state and time-resolved nucleotide binding to dissect the role of NBS2 of the accessory SUR1 subunit of KATP in channel gating. Binding to NBS2 was Mg2+-independent, but Mg was required to trigger a conformational change in SUR1. Mutation of a lysine (K1384A) in NBS2 that coordinates bound nucleotides increased the EC50 for trinitrophenyl-ADP binding to NBS2, but only in the presence of Mg2+, indicating that this mutation disrupts the ligand-induced conformational change. Comparison of nucleotide-binding with ionic currents suggests a model in which each nucleotide binding event to NBS2 of SUR1 is independent and promotes KATP activation by the same amount.

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