The modulator protein dissociates the catalytic subunit of hepatic protein phosphatase G from glycogen.
1. The phosphorylase phosphatase and glycogen-synthase phosphatase activities associated with the glycogen particles from rat liver were progressively inhibited by incubation with modulator protein. However, the phosphorylase phosphatase activity of the catalytic subunit was entirely recovered after destruction of the modulator and the regulatory subunit(s) by trypsin. 2. Inhibition of protein phosphatase G by modulator was associated with a translocation of the phosphorylase phosphatase activity (measured after incubation with trypsin) from glycogen to the soluble fraction. The degree of inhibition of phosphatase G corresponded closely to the extent to which the phosphorylase phosphatase activity was released from the glycogen particles. Incubation of glycogen-free protein phosphatase G with modulator did not change the affinity of the enzyme for added glycogen, but decreased the amount of phosphatase that could be bound to glycogen. 3. The phosphorylase phosphatase activity that was released from the glycogen particles by modulator migrated on gel filtration as a complex (Mr 106,000) of the catalytic subunit with modulator. Phosphorylase phosphatase activity could be transferred from glycogen-bound protein phosphatase G to modulator that was covalently bound to Sepharose. After elution from the column, the enzyme was identified as the free catalytic subunit (Mr 37,000).
Bollen
M
M
Afdeling Biochemie, Faculteit Geneeskunde, Katholieke Universiteit Leuven, Belgium.
Stalmans
W
W
D000818:Animals; D002384:Catalysis; D002850:Chromatography, Gel; D006007:Glycogen-Synthase-D Phosphatase / Q000037:antagonists & inhibitors* / Q000378:metabolism; D066298:In Vitro Techniques; D008099:Liver / Q000187:drug effects / Q000201:enzymology*; D008112:Liver Glycogen / Q000378:metabolism*; D008970:Molecular Weight; D010749:Phosphoprotein Phosphatases / Q000037:antagonists & inhibitors*; D010765:Phosphorylase Phosphatase / Q000378:metabolism; D011485:Protein Binding; D011506:Proteins / Q000494:pharmacology*; D051381:Rats; D013347:Subcellular Fractions / Q000187:drug effects / Q000201:enzymology; D000097847:Protein Phosphatase Inhibitory Proteins
D006007:Glycogen-Synthase-D Phosphatase; D008112:Liver Glycogen; D010749:Phosphoprotein Phosphatases; D010765:Phosphorylase Phosphatase; D011506:Proteins; C024833:protein phosphatase inhibitor-2; D000097847:Protein Phosphatase Inhibitory Proteins