Tyrosine phosphorylation of rod cyclic nucleotide-gated channels switches off Ca2+/calmodulin inhibition

J Neurosci. 2003 Nov 5;23(31):10100-6. doi: 10.1523/JNEUROSCI.23-31-10100.2003.

Abstract

Cyclic nucleotide-gated (CNG) ion channels are crucial for phototransduction in rod photoreceptors. Light triggers a biochemical cascade that reduces the concentration of cGMP in rods, closing CNG channels, which leads to membrane potential hyperpolarization and a decrease in the concentration of intracellular Ca2+. During light adaptation, the sensitivity of CNG channels to cGMP is decreased by Ca2+, which in conjunction with calmodulin (CaM), binds directly to CNG channels. The cGMP sensitivity of rod CNG channels is also reduced by phosphorylation of specific tyrosine residues in the three CNGA1 subunits and one CNGB1 subunit that comprise the rod channel. Here we show that phosphorylation prevents Ca2+/CaM inhibition. Experiments on native channels in rod outer segments and expressed channels in Xenopus oocytes show that Ca2+/CaM inhibition can be toggled off or on by promoting phosphorylation or dephosphorylation, respectively. Experiments in which the crucial tyrosine phosphorylation sites in CNGA1 and CNGB1 are replaced with phenylalanines show that residue Y498 in CNGA1 is the phosphorylation site responsible for regulating Ca2+/CaM inhibition. Ca2+/CaM inhibits the rod channel by binding to the N terminus of the CNGB1 subunit, causing it to uncouple from the C terminus of CNGA1. We propose that phosphorylation of CNGA1Y498, on the C terminus of CNGA1, triggers an equivalent uncoupling from the C terminus of CNGB1, thereby curtailing Ca2+/CaM inhibition. The control of CaM inhibition by CNG channel phosphorylation may be important for light adaptation and the regulation of phototransduction by IGF-1, a retinal paracrine factor that alters the tyrosine phosphorylation state of rod CNG channels.

Publication types

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

MeSH terms

  • Animals
  • Calcium / metabolism*
  • Calmodulin / metabolism*
  • Cattle
  • Cyclic Nucleotide-Gated Cation Channels
  • Eye Proteins / drug effects
  • Eye Proteins / genetics
  • Eye Proteins / metabolism
  • Ion Channel Gating / drug effects
  • Ion Channel Gating / physiology
  • Ion Channels / drug effects
  • Ion Channels / genetics
  • Ion Channels / metabolism*
  • Microinjections
  • Mutation
  • Oocytes / drug effects
  • Oocytes / metabolism
  • Patch-Clamp Techniques
  • Phosphorylation / drug effects
  • RNA, Complementary / genetics
  • Retinal Rod Photoreceptor Cells / metabolism*
  • Tyrosine / metabolism*
  • Vanadates / pharmacology
  • Xenopus laevis

Substances

  • CNGA1 protein, bovine
  • Calmodulin
  • Cyclic Nucleotide-Gated Cation Channels
  • Eye Proteins
  • Ion Channels
  • RNA, Complementary
  • pervanadate
  • Vanadates
  • Tyrosine
  • Calcium