ABSTRACT
Phosphatidate phosphohydrolase (PAPase) and diacylglycerol lipase (DGL) enzymatic activities were found to be differently affected by preincubation of rod outer segments (ROS) under protein phosphorylation or dephosphorylation conditions in darkness or in light. Under protein kinase C (PKC) phosphorylation conditions, PAPase and DGL were inhibited in darkness and in light. The inhibitory effect on PAPase and DGL activities by PKC phosphorylation in the presence of light was more pronounced when the activities were compared with the activities in control membranes determined in the presence of EGTA. The addition of PKC activators such as phorbol-12,13-dibutyrate and dioctanoylglycerol (DOG) instead of DG produced the same pattern of changes in enzymatic activities. Pretreatment of ROS membranes with cAMP-dependent protein kinase (PKA) produced a significant increase in both enzymatic activities in the presence of light. No changes were observed when ROS proteins were phosphorylated by PKA in the dark. Dephosphorylation of ROS membranes with alkaline phosphatase resulted in a decrease in PAPase activity that was more marked under light than under dark conditions. DGL activity was not modified under dephosphorylation conditions. These findings suggest that the metabolization of phosphatidic acid in isolated ROS is differently affected by protein phosphorylation and dephosphorylation reactions.
