Interaction sites between the Slo1 pore and the NH2 terminus of the beta2 subunit, probed with a three-residue sensor

J Biol Chem. 2007 Jun 15;282(24):17720-8. doi: 10.1074/jbc.M607063200. Epub 2007 Apr 12.

Abstract

Calcium- and voltage-gated (BK) K(+) channels encoded by Slo1 play an essential role in nervous systems. Although it shares many common features with voltage-dependent K(V) channels, the BK channel exhibits differences in gating and inactivation. Using a mutant in which FWI replaces three residues (FIW) in the NH(2) terminus of wild-type beta2-subunits, in conjunction with alanine-scanning mutagenesis of the Slo1 S6 segment, we identify that the NH(2) terminus of beta2-subunits interacts with the residues near the cytosolic superficial mouth of BK channels during inactivation. The cytosolic blockers did not share the sites with NH(2) terminus of beta2-subunits. A novel blocking-inactivating scheme was proposed to account for the observed non-competition inactivation. Our results also suggest that the residue Ile-323 plays a dual role in interacting with the NH(2) terminus of beta2-subunits and modulating the gating of BK channels.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Amino Acid Sequence
  • Animals
  • Binding Sites
  • DNA Mutational Analysis
  • Humans
  • Large-Conductance Calcium-Activated Potassium Channel alpha Subunits / chemistry
  • Large-Conductance Calcium-Activated Potassium Channel alpha Subunits / genetics
  • Large-Conductance Calcium-Activated Potassium Channel alpha Subunits / metabolism*
  • Large-Conductance Calcium-Activated Potassium Channel beta Subunits* / chemistry
  • Large-Conductance Calcium-Activated Potassium Channel beta Subunits* / genetics
  • Large-Conductance Calcium-Activated Potassium Channel beta Subunits* / metabolism
  • Mice
  • Models, Molecular
  • Molecular Sequence Data
  • Molecular Structure
  • Mutagenesis, Site-Directed
  • Patch-Clamp Techniques
  • Protein Structure, Tertiary*
  • Sequence Alignment
  • Xenopus laevis

Substances

  • Kcnma1 protein, mouse
  • Large-Conductance Calcium-Activated Potassium Channel alpha Subunits
  • Large-Conductance Calcium-Activated Potassium Channel beta Subunits