Enhancement by phosphodiesterase subunits of the rate of GTP hydrolysis by transducin in bovine retinal rods. Essential role of the phosphodiesterase catalytic core.
Phosphodiesterase (PDE) in bovine retinal rod outer segments is activated when it forms a membrane-bound complex with the alpha-subunit of transducin loaded with GTP (T alpha*). At maximal activation, this complex contains two T alpha* and all the subunits of native PDE (PDE alpha, PDE beta, and two inhibitory PDE gamma). We observed previously (Pagès, F., Deterre, P., and Pfister, C. (1992) J. Biol. Chem. 267, 22018-22021) that the rate of GTP hydrolysis by transducin in a rod outer segment suspension is enhanced when T alpha* is bound to native PDE (PDE alpha beta gamma 2). In this article, we compare the effects of PDE species with different PDE gamma contents. We show that T alpha* hydrolyzes its GTP faster not only when bound to PDE alpha beta gamma 2, but also when bound to PDE alpha beta gamma or PDE alpha beta. Moreover, trypsin-treated PDE (PDE gamma-deprived soluble PDE) also induces an acceleration of GTP hydrolysis. On the contrary, addition of isolated PDE gamma alone does not accelerate GTP hydrolysis. The interaction between T alpha* and PDE gamma, which is essential for the activation of PDE by T alpha*, is apparently not responsible of the feedback of PDE on T alpha*. The interaction of primary importance for the acceleration of GTP hydrolysis would be that existing between T alpha* and PDE alpha beta.
Pagès
F
F
Département de Biologie Moléculaire et Structurale, Centre d'Etudes Nucléaires de Grenoble, France.
Deterre
P
P
Pfister
C
C
D000818:Animals; D002384:Catalysis; D002417:Cattle; D006160:Guanosine Triphosphate / Q000378:metabolism*; D006868:Hydrolysis; D007700:Kinetics; D008565:Membrane Proteins / Q000378:metabolism; D010727:Phosphoric Diester Hydrolases / Q000378:metabolism*; D017948:Retinal Rod Photoreceptor Cells / Q000378:metabolism*; D015293:Transducin / Q000378:metabolism*
D008565:Membrane Proteins; D006160:Guanosine Triphosphate; D010727:Phosphoric Diester Hydrolases; D015293:Transducin