Reciprocal control of retinal rod cyclic GMP phosphodiesterase by its gamma subunit and transducin.
The switching on of the cGMP phosphodiesterase (PDE) in retinal rod outer segments by activated transducin (T alpha-GTP) is a key step in visual excitation. The finding that trypsin activates PDE (alpha beta gamma) by degrading its gamma subunit and the reversal of this activation by gamma led to the proposal that T alpha-GTP activates PDE by relieving an inhibitory constraint imposed by gamma (Hurley and Stryer: J. Biol. Chem. 257:11094-11099, 1982). We report here studies showing that the addition of gamma subunit also reverses the activation of PDE by T alpha-GTP-gamma S. A procedure for preparing gamma in high yield (50-80%) is presented. Analyses of SDS polyacrylamide gel slices confirmed that inhibitory activity resides in the gamma subunit. Nanomolar gamma blocks the activation of PDE by micromolar T alpha-GTP gamma S. The degree of activation of PDE depends reciprocally on the concentrations of gamma and T alpha-GTP gamma S. gamma remains bound to the disk membrane during the activation of PDE by transducin. The binding of gamma to the alpha beta subunits of native PDE is very tight; the dissociation constant is less than 10 pM, indicating that fewer than 1 in 1,700 PDE molecules in rod outer segments are activated in the absence of T alpha-GTP.
Wensel
T G
TG
Department of Cell Biology, Sherman Fairchild Center, Stanford University School of Medicine, California 94305.
Stryer
L
L
D015106:3',5'-Cyclic-GMP Phosphodiesterases / Q000378:metabolism*; D000818:Animals; D002417:Cattle; D004789:Enzyme Activation / Q000187:drug effects / Q000528:radiation effects; D066298:In Vitro Techniques; D008027:Light; D008565:Membrane Proteins / Q000494:pharmacology*; D010786:Photoreceptor Cells / Q000201:enzymology*; D011487:Protein Conformation; D012374:Rod Cell Outer Segment / Q000201:enzymology*; D015293:Transducin; D014357:Trypsin / Q000494:pharmacology
D008565:Membrane Proteins; D015106:3',5'-Cyclic-GMP Phosphodiesterases; D014357:Trypsin; D015293:Transducin