ATP-sensitive K+ channels: Regulation of bursting by the sulphonylurea receptor, PIP2 and regions of Kir6.2

Bernard Ribalet, Scott A. John, Lai Hua Xie, James N. Weiss

Research output: Contribution to journalArticlepeer-review

26 Scopus citations

Abstract

ATP-sensitive K+ channels composed of the pore-forming protein Kir6.2 and the sulphonylurea receptor SUR1 are inhibited by ATP and activated by Phosphatidylinositol Bisphosphate (PIP2). Residues involved in binding of these ligands to the Kir6.2 cytoplasmic domain have been identified, and it has been hypothesized that gating mechanisms involve conformational changes in the regions of the bundle crossing and/or the selectivity filter of Kir6.2. Regulation of Kir6.2 by SUR1, however, is not well-understood, even though this process is ATP and PIP2 dependent. In this study, we investigated the relationship between channel regulation by SUR1 and PIP2 by comparing a number of single and double mutants known to affect open probability (Po), PIP2 affinity, and sulphonylurea and MgADP sensitivity. When coexpressed with SUR1, the Kir6.2 mutant C166A, which is characterized by a Po value close to 0.8, exhibits no sulphonylurea or MgADP sensitivity. However, when Po was reduced by combiningmutations at the PIP2 -sensitive residues R176 and R177 with C166A, sulphonylurea and MgADP sensitivities were restored. These effects correlated with a dramatic decrease in PIP2 affinity, as assessed by PIP2 -induced channel reactivation and inhibition by neomycin, an antagonist of PIP2 binding. Based on macroscopic and single-channel data, we propose a model in which entry into the high-Po bursting state by the C166A mutation or by SUR1 depends on the interaction of PIP2 with R176 and R177 and, to a lesser extent, R54. In conjunction with this PIP2-dependent process, SUR1 also regulates channel activity via a PIP2-independent, but MgADP-dependent process.

Original languageEnglish (US)
Pages (from-to)303-317
Number of pages15
JournalJournal of Physiology
Volume571
Issue number2
DOIs
StatePublished - Mar 2006
Externally publishedYes

All Science Journal Classification (ASJC) codes

  • Physiology

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