Properties and content of cyclic nucleotide phosphodiesterase in photoreceptor outer segments of ground squirrel retina.
Cyclic GMP-specific phosphodiesterase (3',5'-cyclic-nucleotide 5'-nucleotidohydrolase, EC 1.3.4.17) (PDE) is thought to be a key enzyme of the retinal-rod phototransduction cyclic nucleotide pathway. We attempted to investigate the properties and content of PDE in retinal-cone photoreceptors. The fractions obtained from cone-dominant ground squirrel retinas were analyzed for cone visual pigment content and PDE activity. The cone visual pigment content was estimated to be approx. 65 pmol per retina. The distribution of cone visual pigment coincided with that of the PDE activity through several steps of photoreceptor membrane purification by sucrose density gradient centrifugation. The ground squirrel retinal PDE was similar to the retinal-rod PDE by its kinetic properties, thermostability, sensitivity to tryptic activation, Stokes radius and pI values. The cone visual pigment enriched fractions contained the heat-stable trypsin-inactivated PDE inhibitor. Its functional properties seem to be similar to those of the retinal-rod PDE inhibitory subunit. The PDE content in ground squirrel retina was roughly estimated to be about five copies of enzyme per 100 cone visual pigment molecules. The obtained results indicated that the major portion of ground squirrel retinal cyclic GMP-specific PDE is the endogenous cone photoreceptor membrane enzyme and strongly supported the conception about the key role of PDE in cone phototransduction. The existence of essential differences between rod and cone systems rapidly returning cyclic GMP-specific amplification cascade components to the dark (or inactivated) states after photon absorption was suggested. If this suggestion is true, the well-known distinctions between response kinetics and light sensitivity of these two kinds of photoreceptor can be explained.
Orlov NYa
Institute of Biological Physics, U.S.S.R. Academy of Sciences, Pushchino, Moscow Region.
Kalinin
E V
EV
Orlova
T G
TG
Freidin
A A
AA
D015106:3',5'-Cyclic-GMP Phosphodiesterases / Q000037:antagonists & inhibitors / Q000502:physiology*; D000818:Animals; D002458:Cell Fractionation; D007700:Kinetics; D010786:Photoreceptor Cells / Q000201:enzymology*; D012589:Sciuridae; D013057:Spectrum Analysis
D015106:3',5'-Cyclic-GMP Phosphodiesterases