Molecular organization of bovine rod cGMP-phosphodiesterase 6

J Mol Biol. 2001 Jul 20;310(4):781-91. doi: 10.1006/jmbi.2001.4813.

Abstract

Phosphodiesterase 6 (PDE6), a multisubunit (alphabetagamma(2)delta) enzyme, plays a major role in visual function by hydrolysing cGMP in response to a light stimulus. Solubilized bovine rod PDE6 molecules depleted of their gamma subunits were purified to homogeneity from bovine retinal rods and their molecular organization was investigated by electron microscopy. Image analysis of single particles revealed the three-dimensional dimeric arrangement of the purified alphabetadelta complex, and the internal organization of each catalytic subunit into three distinct domains at a resolution of 2.8 nm. The relative volume of each domain is consistent with sequence analysis and functional data, which suggest that these domains correspond to the catalytic and two GAF domains. This hypothesis was confirmed by immunolabelling experiments, which located the N-terminal part of the catalytic subunit where the major interaction between the two alphabeta subunits was found to occur. The 3D molecular organization of human platelet PDE5 appears highly homologous to that of bovine rod PDE6, as predicted by similarities in their primary sequences. These observations describe the quaternary organization of the catalytic PDE6 alphabeta complex, and place the catalytic and regulatory domains on a structural model.

Publication types

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

MeSH terms

  • 3',5'-Cyclic-GMP Phosphodiesterases / chemistry*
  • 3',5'-Cyclic-GMP Phosphodiesterases / isolation & purification
  • 3',5'-Cyclic-GMP Phosphodiesterases / metabolism
  • 3',5'-Cyclic-GMP Phosphodiesterases / ultrastructure*
  • Animals
  • Blood Platelets / enzymology
  • Catalytic Domain
  • Cattle
  • Cyclic Nucleotide Phosphodiesterases, Type 5
  • Cyclic Nucleotide Phosphodiesterases, Type 6
  • Humans
  • Microscopy, Electron
  • Microscopy, Immunoelectron
  • Phosphoric Diester Hydrolases / chemistry
  • Phosphoric Diester Hydrolases / isolation & purification
  • Phosphoric Diester Hydrolases / metabolism
  • Phosphoric Diester Hydrolases / ultrastructure
  • Protein Structure, Quaternary
  • Protein Subunits
  • Retinal Rod Photoreceptor Cells / enzymology*

Substances

  • Protein Subunits
  • Phosphoric Diester Hydrolases
  • 3',5'-Cyclic-GMP Phosphodiesterases
  • Cyclic Nucleotide Phosphodiesterases, Type 5
  • Cyclic Nucleotide Phosphodiesterases, Type 6
  • PDE5A protein, human
  • PDE6B protein, human