Differential activities of protein phosphatase types 1 and 2A in cytosolic and particulate fractions from rat forebrain.
The activities and concentrations of protein phosphatase type 1 (PP1) and type 2A (PP2A) were compared in cytosol and particulate fractions of rat forebrain. Although the activity of PP2A was highest in the cytosol, immunoblot analysis with a PP2A-specific antibody showed that there were significant levels of the enzyme in the particulate fraction. There was no significant difference between the concentration of PP2A in the cytosol and particulate fractions such that the low activity of PP2A in the particulate fraction represents an inactivation of this form of the enzyme. Similar analysis in skeletal muscle, heart, and liver showed this finding was unique to the brain. Similarly, the majority of PP1 activity was recovered in the cytosol, but most PP1 enzyme was associated with the particulate fraction. Comparison with other tissues showed that the activities of PP1 in the particulate fractions were similar but that the forebrain contained significantly more enzyme than the other tissues. Thus, like PP2A it appears that the specific activity of PP1 in the particulate fraction of rat forebrain is much lower than that of the cytosol and of the particulate fractions of other tissues. Elution of PP1 and PP2A from membranes with 0.5 M NaCl plus 0.3% Triton X-100 resulted in severalfold activation of both enzymes. That the majority of PP1 and PP2A in rat forebrain are associated with membrane structures but in a low activity state suggests that novel regulatory mechanisms exist that have considerable and unique potential for activation of protein dephosphorylation.
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Neuroscience Group, University of Newcastle, Callaghan, New South Wales, Australia.
Ratcliffe
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Mumby
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Villa-Moruzzi
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Rostas
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JA
D000818:Animals; D002352:Carrier Proteins; D002462:Cell Membrane / Q000201:enzymology; D003600:Cytosol / Q000201:enzymology; D004988:Ethers, Cyclic / Q000494:pharmacology; D015151:Immunoblotting; D047908:Intracellular Signaling Peptides and Proteins; D008099:Liver / Q000201:enzymology / Q000648:ultrastructure; D008297:Male; D009132:Muscles / Q000201:enzymology / Q000648:ultrastructure; D009206:Myocardium / Q000201:enzymology / Q000648:ultrastructure; D019319:Okadaic Acid; D010749:Phosphoprotein Phosphatases / Q000037:antagonists & inhibitors / Q000378:metabolism*; D016548:Prosencephalon / Q000201:enzymology* / Q000648:ultrastructure*; D011506:Proteins / Q000494:pharmacology; D051381:Rats; D017208:Rats, Wistar
D002352:Carrier Proteins; D004988:Ethers, Cyclic; D047908:Intracellular Signaling Peptides and Proteins; D011506:Proteins; C021042:protein phosphatase inhibitor-1; D019319:Okadaic Acid; D010749:Phosphoprotein Phosphatases