The pore region of the Kv1.2alpha subunit is an important component of recombinant Kv1.2 channel oxygen sensitivity

Biochem Biophys Res Commun. 2003 Jun 27;306(2):450-6. doi: 10.1016/s0006-291x(03)00989-6.

Abstract

Oxygen-sensitive K(+) channels are important elements in the cellular response to hypoxia. Although much progress has been made in identifying their molecular composition, the structural components associated to their O(2)-sensitivity are not yet understood. Recombinant Kv1.2 currents expressed in Xenopus oocytes are inhibited by a decrease in O(2) availability. On the contrary, heterologous Kv2.1 channels are O(2)-insensitive. To elucidate the protein segment responsible for the O(2)-sensitivity of Kv1.2 channels, we analyzed the response to anoxia of Kv1.2/Kv2.1 chimeric channels. Expression of chimeric Kv2.1 channels each containing the S4, the S1-S3 or the S6-COOH segments of Kv1.2 polypeptide resulted in a K(+) current insensitive to anoxia. In contrast, transferring the S5-S6 segment of Kv1.2 into Kv2.1 produced an O(2)-sensitive K(+) current. Finally, mutating a redox-sensitive methionine residue (M380) of Kv1.2 polypeptide did not affect O(2)-sensitivity. Thus, the pore and its surrounding regions of Kv1.2 polypeptide confer its hypoxic inhibition. This response is independent on the redox modulation of methionine residues in this protein segment.

Publication types

  • Research Support, Non-U.S. Gov't
  • Research Support, U.S. Gov't, Non-P.H.S.
  • Research Support, U.S. Gov't, P.H.S.

MeSH terms

  • Amino Acid Sequence
  • Animals
  • Electrophysiology
  • Hypoxia
  • Kv1.2 Potassium Channel
  • Molecular Sequence Data
  • Oocytes / metabolism
  • Oxidation-Reduction
  • Oxygen / metabolism
  • Peptides / chemistry
  • Point Mutation
  • Potassium / metabolism
  • Potassium Channels / chemistry*
  • Potassium Channels / metabolism
  • Potassium Channels, Voltage-Gated*
  • Protein Structure, Tertiary
  • RNA, Complementary / metabolism
  • Recombinant Proteins / chemistry*
  • Recombinant Proteins / metabolism
  • Sequence Homology, Amino Acid
  • Xenopus

Substances

  • Kv1.2 Potassium Channel
  • Peptides
  • Potassium Channels
  • Potassium Channels, Voltage-Gated
  • RNA, Complementary
  • Recombinant Proteins
  • Potassium
  • Oxygen