Identification of gamma-aminobutyric acid receptor-interacting factor 1 (TRAK2) as a trafficking factor for the K+ channel Kir2.1

J Biol Chem. 2006 Oct 6;281(40):30104-11. doi: 10.1074/jbc.M602439200. Epub 2006 Aug 8.

Abstract

To identify proteins that regulate potassium channel activity and expression, we performed functional screening of mammalian cDNA libraries in yeast that express the mammalian K(+) channel Kir2.1. Growth of Kir2.1-expressing yeast in media with low K(+) concentration is a function of K(+) uptake via Kir2.1 channels. Therefore, the host strain was transformed with a human cDNA library, and cDNA clones that rescued growth at low K(+) concentration were selected. One of these clones was identical to the protein of unknown function isolated previously as gamma-aminobutyric acid receptor-interacting factor 1 (GRIF-1) (Beck, M., Brickley, K., Wilkinson, H., Sharma, S., Smith, M., Chazot, P., Pollard, S., and Stephenson, F. (2002) J. Biol. Chem. 277, 30079-30090). GRIF-1 specifically enhanced Kir2.1-dependent growth in yeast and Kir2.1-mediated (86)Rb(+) efflux in HEK293 cells. Quantitative microscopy and flow cytometry analysis of immunolabeled surface Kir2.1 channel showed that GRIF-1 significantly increased the number of Kir2.1 channels in the plasma membrane of COS and HEK293 cells. Physical interaction of Kir2.1 channel and GRIF-1 was demonstrated by co-immunoprecipitation from HEK293 lysates and yeast two-hybrid assay. In vivo association of Kir2.1 and GRIF-1 was demonstrated by co-immunoprecipitation from brain lysate. Yeast two-hybrid assays showed that an N-terminal region of GRIF-1 interacts with a C-terminal region of Kir2.1. These results indicate that GRIF-1 binds to Kir2.1 and facilitates trafficking of this channel to the cell surface.

Publication types

  • Research Support, N.I.H., Extramural
  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • COS Cells
  • Carrier Proteins / isolation & purification
  • Carrier Proteins / metabolism
  • Carrier Proteins / physiology*
  • Cell Line
  • Cell Membrane / metabolism
  • Chlorocebus aethiops
  • Humans
  • Intracellular Signaling Peptides and Proteins
  • Nerve Tissue Proteins / isolation & purification
  • Nerve Tissue Proteins / metabolism
  • Nerve Tissue Proteins / physiology*
  • Potassium Channels, Inwardly Rectifying / metabolism*
  • Protein Transport / physiology
  • Saccharomyces cerevisiae / genetics
  • Two-Hybrid System Techniques

Substances

  • Carrier Proteins
  • Intracellular Signaling Peptides and Proteins
  • KCNJ2 protein, human
  • Nerve Tissue Proteins
  • Potassium Channels, Inwardly Rectifying
  • TRAK2 protein, human