Molecular organization of bovine rod cGMP-Phosphodiesterase 6.

Abstract

Phosphodiesterase 6 (PDE6), a multisubunit (αβγ2δ) 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 γ 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 αβδ 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 αβ 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 αβ complex, and place the catalytic and regulatory domains on a structural model.

Department

Molecular, Cellular and Biomedical Sciences

Publication Date

6-20-2001

Journal Title

Journal of Molecular Biology

Publisher

Springer

Digital Object Identifier (DOI)

10.1006/jmbi.2001.4813

Document Type

Article

Rights

© 2001 Academic Press.

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