cGMP binding sites on photoreceptor phosphodiesterase: role in feedback regulation of visual transduction.
A central step in vertebrate visual transduction is the rapid drop in cGMP levels that causes cGMP-gated ion channels in the photoreceptor cell membrane to close. It has long been a puzzle that the cGMP phosphodiesterase (PDE) whose activation causes this decrease contains not only catalytic sites for cGMP hydrolysis but also noncatalytic cGMP binding sites. Recent work has shown that occupancy of these noncatalytic sites slows the rate of PDE inactivation. We report here that PDE activation induced by activated transduction lowers the cGMP binding affinity for noncatalytic sites on PDE and accelerates the dissociation of cGMP from these sites. These sites can exist in three states: high affinity (Kd = 60 nM) for the nonactivated PDE, intermediate affinity (Kd approximately 180 nM) when the enzyme is activated in a complex with transducin, and low affinity (Kd > 1 microM) when transducin physically removes the inhibitory subunits of PDE from the PDE catalytic subunits. Activation of PDE by transducin causes a 10-fold increase in the rate of cGMP dissociation from one of the two noncatalytic sites; physical removal of the inhibitory subunits from the PDE catalytic subunits further accelerates the cGMP dissociation rate from both sites > 50-fold. Because PDE molecules lacking bound cGMP inactivate more rapidly, this suggests that a prolonged cGMP decrease may act as a negative feedback regulator to generate the faster, smaller photoresponses characteristic of light-adapted photoreceptors.
Cote
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RH
Department of Biochemistry and Molecular Biology, University of New Hampshire, Durham 03824.
Bownds
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MD
Arshavsky
V Y
VY
D000818:Animals; D001665:Binding Sites; D002384:Catalysis; D006152:Cyclic GMP / Q000378:metabolism*; D004789:Enzyme Activation; D005246:Feedback; D010727:Phosphoric Diester Hydrolases / Q000378:metabolism*; D011892:Rana catesbeiana; D011898:Ranidae; D012374:Rod Cell Outer Segment / Q000201:enzymology*; D015398:Signal Transduction; D015293:Transducin / Q000378:metabolism; D014795:Visual Pathways / Q000378:metabolism
D010727:Phosphoric Diester Hydrolases; D015293:Transducin; D006152:Cyclic GMP