A functional link between the dark Mg‐ATPase activity and the light‐induced enzymatic cascade in rod outer segments

Abstract

The characterization of a light-induced scattering change in suspensions of rod fragments, which requires previous swelling of the disks by the dark Mg/ATPase described by Uhl et al. [FEBS Lett. 107, 317-322 (1979)] is reported here. Reconstitution experiments demonstrate that this signal is dependent on the presence of G-protein, GTP and cGMP phosphodiesterase. Fast reversal associated with regenerability requires in addition the presence of some protein(s) of the cytoplasm (probably the rhodopsin kinase) and ATP. The amount of excited rhodopsin which saturates the signal is the same as that which saturates the previously described ''dissociation signal'' [Kuhn et al. (1981) Proc. Natl. Acad. Sci. USA 78, 6873-6877] associated with the formation of the phosphodiesterase activator G.alpha.GTP (.alpha. subunit of the G-protein with GTP bound). The kinetics of the signal is slightly slower than that of the dissociation signal and its amplitude is proportional to the extent of swelling of the disks. These results suggest that the interaction between G.alpha.GTP and the phosphodiesterase modifies some structural feature of the disks and provide evidence for the existence of a functional link between the dark Mg-ATPase and the light-induced enzymatic cascade.