The glutamic acid-rich protein-2 (GARP2) is a high affinity rod photoreceptor phosphodiesterase (PDE6)-binding protein that modulates its catalytic properties

J Biol Chem. 2006 Mar 3;281(9):5500-5. doi: 10.1074/jbc.M507488200. Epub 2006 Jan 3.

Abstract

The glutamic acid-rich protein-2 (GARP2) is a splice variant of the beta-subunit of the cGMP-gated ion channel of rod photoreceptors. GARP2 is believed to interact with several membrane-associated phototransduction proteins in rod photoreceptors. In this study, we demonstrated that GARP2 is a high affinity PDE6-binding protein and that PDE6 co-purifies with GARP2 during several stages of chromatographic purification. We found that hydrophobic interaction chromatography succeeds in quantitatively separating GARP2 from the PDE6 holoenzyme. Furthermore, the 17-kDa prenyl-binding protein, abundant in retinal cells, selectively released PDE6 (but not GARP2) from rod outer segment membranes, demonstrating the specificity of the interaction between GARP2 and PDE6. Purified GARP2 was able to suppress 80% of the basal activity of the nonactivated, membrane-bound PDE6 holoenzyme at concentrations equivalent to its endogenous concentration in rod outer segment membranes. However, GARP2 was unable to reverse the transducin activation of PDE6 (in contrast to a previous study) nor did it significantly alter catalysis of the fully activated PDE6 catalytic dimer. The high binding affinity of GARP2 for PDE6 and its ability to regulate PDE6 activity in its dark-adapted state suggest a novel role for GARP2 as a regulator of spontaneous activation of rod PDE6, thereby serving to lower rod photoreceptor "dark noise" and allowing these sensory cells to operate at the single photon detection limit.

Publication types

  • Research Support, N.I.H., Extramural

MeSH terms

  • 3',5'-Cyclic-GMP Phosphodiesterases / chemistry
  • 3',5'-Cyclic-GMP Phosphodiesterases / metabolism*
  • Animals
  • Cattle
  • Cell Membrane / chemistry
  • Cell Membrane / metabolism
  • Cyclic Nucleotide Phosphodiesterases, Type 6
  • Cyclic Nucleotide-Gated Cation Channels
  • Darkness
  • Enzyme Activation
  • Eye Proteins / chemistry
  • Eye Proteins / metabolism*
  • Nerve Tissue Proteins / chemistry
  • Nerve Tissue Proteins / isolation & purification
  • Nerve Tissue Proteins / metabolism*
  • Protein Binding
  • Protein Isoforms / chemistry
  • Protein Isoforms / metabolism
  • Protein Subunits / chemistry
  • Protein Subunits / metabolism
  • Retina / chemistry
  • Retina / cytology
  • Retina / metabolism*
  • Retinal Rod Photoreceptor Cells / cytology
  • Retinal Rod Photoreceptor Cells / enzymology*
  • Rod Cell Outer Segment / chemistry
  • Rod Cell Outer Segment / metabolism
  • Transducin / chemistry
  • Transducin / metabolism

Substances

  • CNGB1 protein, Bos taurus
  • Cyclic Nucleotide-Gated Cation Channels
  • Eye Proteins
  • Nerve Tissue Proteins
  • Protein Isoforms
  • Protein Subunits
  • 3',5'-Cyclic-GMP Phosphodiesterases
  • Cyclic Nucleotide Phosphodiesterases, Type 6
  • Transducin