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Purification and partial characterization of glycogen synthase kinase-3 from rabbit liver

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Summary

Glycogen synthase kinase-3 (GSK-3) was purified from rabbit liver to homogeneity by ultracentrifugation, ion-exchange chromatography on DEAE-cellulose, Cellulose phosphate, CM-Sephadex and Fast Protein Liquid Chromatography (FPLC) on Mono-S column. The enzyme was purified approximately 20,000 fold with an approximate 2% recovery. The purified enzyme showed a single band on SDS-polyacrylamide gel electrophoresis. GSK-3 is a monomeric enzyme with a molecular weight of 50,000–52,000 as derived from SDS-polyacrylamide gel electrophoresis and gel filtration. The purified enzyme was indeed a GSK-3 since it phosphorylated three sites, i.e., 3a, 3b, and 3c on liver glycogen synthase. GSK-3 incorporated up to 2.6 mol Pi/mol glycogen synthase subunit with a concomitant inactivation of glycogen synthase activity.

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Correspondence to Ramji L. Khandelwal.

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Randhawa, R., Khandelwal, R.L. Purification and partial characterization of glycogen synthase kinase-3 from rabbit liver. Mol Cell Biochem 95, 147–155 (1990). https://doi.org/10.1007/BF00219973

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Key words

  • glycogen synthase kinase-3
  • glycogen synthase
  • rabbit liver
  • protein enzyme purification
  • protein kinases
  • glycogen