Direct activation of cGMP-dependent channels of retinal rods by the cGMP phosphodiesterase.
The cationic conductances of purified bovine retinal rod membranes were studied by incorporation of vesicles into planar lipid bilayers. When the membranes were stripped of all peripheral proteins [guanine nucleotide-binding protein (G protein) and cGMP phosphodiesterase (3',5'-cyclic-GMP 5'-nucleotidohydrolase), EC 3.1.4.35], sodium and calcium fluxes were almost only observed in the presence of cGMP. Reconstitution experiments in which purified cGMP phosphodiesterase alone or with G protein were reassociated to the vesicles in proportions similar to those found in the native rod provide evidence for a direct interaction between the cGMP-dependent channel protein and the phosphodiesterase. (i) In its inhibited state, phosphodiesterase markedly stimulates the activity of the channels in the presence of cGMP (situation in the dark-adapted rod) but is not capable of activating the channels in the absence of cGMP. (ii) In the absence of cGMP, activation of the phosphodiesterase by G protein with GTP bound (equivalent to photoexcitation) induces the opening of cation channels that have the same conductance for sodium ions as cGMP-activated channels (20-22 pS, with two sublevels of about 7 pS and 13 pS).
Bennett
N
N
Centre National de la Recherche Scentifique, URA 520, Centre d'Etudes Nucléaires de Grenoble, France.
Ildefonse
M
M
Crouzy
S
S
Chapron
Y
Y
Clerc
A
A
D015106:3',5'-Cyclic-GMP Phosphodiesterases / Q000502:physiology*; D000818:Animals; D002417:Cattle; D006152:Cyclic GMP / Q000502:physiology*; D004110:Diltiazem / Q000494:pharmacology; D004553:Electric Conductivity; D019204:GTP-Binding Proteins / Q000502:physiology*; D066298:In Vitro Techniques; D007473:Ion Channels / Q000502:physiology*; D008564:Membrane Potentials; D008970:Molecular Weight; D010786:Photoreceptor Cells / Q000502:physiology*; D012964:Sodium / Q000502:physiology
D007473:Ion Channels; D012964:Sodium; D015106:3',5'-Cyclic-GMP Phosphodiesterases; D019204:GTP-Binding Proteins; D004110:Diltiazem; D006152:Cyclic GMP