Can phosphorylation and dephosphorylation of rod outer segment membranes affect phosphatidate phosphohydrolase and diacylglycerol lipase activities?
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.
Pérez Roque
M E
ME
Instituto de Investigaciones Bioquímicas, Universidad Nacional del Sur y Consejo Nacional de Investigaciones Científicas y Técnicas, Bahía Blanca, Argentina.
Pasquaré
S J
SJ
Castagnet
P I
PI
Giusto
N M
NM
D000818:Animals; D002417:Cattle; D017868:Cyclic AMP-Dependent Protein Kinases / Q000378:metabolism; D003624:Darkness; D066298:In Vitro Techniques; D008027:Light; D008071:Lipoprotein Lipase / Q000378:metabolism*; D008565:Membrane Proteins / Q000378:metabolism; D008566:Membranes / Q000378:metabolism / Q000528:radiation effects; D010711:Phosphatidate Phosphatase / Q000378:metabolism*; D010766:Phosphorylation; D012243:Rhodopsin / Q000378:metabolism; D012374:Rod Cell Outer Segment / Q000378:metabolism* / Q000528:radiation effects
D008565:Membrane Proteins; D012243:Rhodopsin; D017868:Cyclic AMP-Dependent Protein Kinases; D008071:Lipoprotein Lipase; D010711:Phosphatidate Phosphatase