The TRPM6 kinase domain determines the Mg·ATP sensitivity of TRPM7/M6 heteromeric ion channels

J Biol Chem. 2014 Feb 21;289(8):5217-27. doi: 10.1074/jbc.M113.512285. Epub 2014 Jan 2.

Abstract

The transient receptor potential melastatin member 7 (TRPM7) and member 6 (TRPM6) are divalent cation channel kinases essential for magnesium (Mg(2+)) homeostasis in vertebrates. It remains unclear how TRPM6 affects divalent cation transport and whether this involves functional homomeric TRPM6 plasma membrane channels or heteromeric channel assemblies with TRPM7. We show that homomeric TRPM6 is highly sensitive to intracellular free Mg(2+) and therefore unlikely to be active at physiological levels of [Mg(2+)]i. Co-expression of TRPM7 and TRPM6 produces heteromeric TRPM7/M6 channels with altered pharmacology and sensitivity to intracellular Mg·ATP compared with homomeric TRPM7. Strikingly, the activity of heteromeric TRPM7/M6 channels is independent of intracellular Mg·ATP concentrations, essentially uncoupling channel activity from cellular energy status. Disruption of TRPM6 kinase phosphorylation activity re-introduces Mg·ATP sensitivity to the heteromeric channel similar to that of TRPM7. Thus, TRPM6 modulates the functionality of TRPM7, and the TRPM6 kinase plays a critical role in tuning the phenotype of the TRPM7·M6 channel complex.

Keywords: Magnesium; Metabolic Regulation; Protein Complexes; Protein Kinases; TRP channels.

Publication types

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

MeSH terms

  • Adenosine Triphosphate / pharmacology*
  • Boron Compounds / pharmacology
  • HEK293 Cells
  • Humans
  • Intracellular Space / drug effects
  • Intracellular Space / metabolism
  • Ion Channel Gating / drug effects
  • Osmolar Concentration
  • Phosphotransferases / metabolism
  • Point Mutation / genetics
  • Protein Structure, Tertiary
  • Protein-Serine-Threonine Kinases
  • Solutions
  • Structure-Activity Relationship
  • TRPM Cation Channels / chemistry*
  • TRPM Cation Channels / metabolism*

Substances

  • Boron Compounds
  • Solutions
  • TRPM Cation Channels
  • TRPM6 protein, human
  • Adenosine Triphosphate
  • 2-aminoethoxydiphenyl borate
  • Phosphotransferases
  • Protein-Serine-Threonine Kinases
  • TRPM7 protein, human