Purification and Characterization of Mg2+-Dependent Glycogen Synthase Phosphatase (Phosphoprotein Phosphatase IA) from Rat Liver

Abstract
Phosphoprotein phosphatase IA, which represents the major glycogen synthase phosphatase activity in rat liver cytosol, has been purified to apparent homogeneity by chromatography on DEAE-cellulose, histone–Sepharose-4B and Sephadex G-100. The molecular weight of the purified enzyme was 40000 by gel filtration and 48000 by sodium dodecyl sulafte gel electrophoresis, phosphatase IA is therefore a monomeric protein. When treated with 80% ethanol at room temperature, phosphatase IA underwent an inactivation which was totally prevented by 2 mM, MgCl2. Catalytically, phosphatase IA has a preference for glycogen synthase D compared with phosphatases IB and II and obligatorily requires Mg2+or Mn2+for activity. Maximum activity was attained at 5 mM Mgcl2. Since Mg2+does not activate other phosphoprotein phosphatases in rat liver cytosol, we propose the term ‘Mg2+-dependent glycogen synthase phosphatase’ for phosphatase IA.

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