Molecular architecture of photoreceptor phosphodiesterase elucidated by chemical cross-linking and integrative modeling.

Academic Article


  • Photoreceptor phosphodiesterase (PDE6) is the central effector enzyme in visual excitation pathway in rod and cone photoreceptors. Its tight regulation is essential for the speed, sensitivity, recovery and adaptation of visual detection. Although major steps in the PDE6 activation/deactivation pathway have been identified, mechanistic understanding of PDE6 regulation is limited by the lack of knowledge about the molecular organization of the PDE6 holoenzyme (αβγγ). Here, we characterize the PDE6 holoenzyme by integrative structural determination of the PDE6 catalytic dimer (αβ), based primarily on chemical cross-linking and mass spectrometric analysis. Our models built from high-density cross-linking data elucidate a parallel organization of the two catalytic subunits, with juxtaposed α-helical segments within the tandem regulatory GAF domains to provide multiple sites for dimerization. The two catalytic domains exist in an open configuration when compared to the structure of PDE2 in the apo state. Detailed structural elements for differential binding of the γ-subunit to the GAFa domains of the α- and β-subunits are revealed, providing insight into the regulation of the PDE6 activation/deactivation cycle.
  • Authors

  • Zeng-Elmore, Xiaohui
  • Gao, Xiong-Zhuo
  • Pellarin, Riccardo
  • Schneidman-Duhovny, Dina
  • Zhang, Xiu-Jun
  • Kozacka, Katie A
  • Tang, Yang
  • Sali, Andrej
  • Chalkley, Robert J
  • Cote, Rick
  • Chu, Feixia
  • Status

    Publication Date

  • November 11, 2014
  • Published In


  • Animals
  • Catalytic Domain
  • Cattle
  • Chromatography, Liquid
  • Cross-Linking Reagents
  • Cyclic Nucleotide Phosphodiesterases, Type 6
  • Eye Proteins
  • Models, Molecular
  • Peptide Fragments
  • Protein Binding
  • Protein Conformation
  • Protein Multimerization
  • Retina
  • Tandem Mass Spectrometry
  • chemical cross-linking
  • integrative modeling
  • mass spectrometry
  • phosphodiesterase 6 (PDE6)
  • visual transduction
  • Digital Object Identifier (doi)

    Start Page

  • 3713
  • End Page

  • 3728
  • Volume

  • 426
  • Issue

  • 22