Purification of GSK-3 by affinity chromatography on immobilized axin

Aline Primot, Blandine Baratte, Marie Gompel, Annie Borgne, Sylvie Liabeuf, Jean Louis Romette, Eek Hoon Jho, Frank Costantini, Laurent Meijer

Research output: Contribution to journalArticlepeer-review

90 Scopus citations


Glycogen synthase kinase 3 (GSK-3), an element of the Wnt signalling pathway, plays a key role in numerous cellular processes including cell proliferation, embryonic development, and neuronal functions. It is directly involved in diseases such as cancer (by controlling apoptosis and the levels of β-catenin and cyclin D1), Alzheimer's disease (tau hyperphosphorylation), and diabetes (as a downstream element of insulin action, GSK-3 regulates glycogen and lipid synthesis). We describe here a rapid and efficient method for the purification of GSK-3 by affinity chromatography on an immobilized fragment of axin. Axin is a docking protein which interacts with GSK-3β, β-catenin, phosphatase 2A, and APC. A polyhistidine-tagged axin peptide (residues 419-672) was produced in Escherichia coli and either immobilized on Ni-NTA agarose beads or purified and immobilized on CNBr-activated Sepharose 4B. These 'Axin-His6' matrices were found to selectively bind recombinant rat GSK-3β and native GSK-3 from yeast, sea urchin embryos, and porcine brain. The affinity-purified enzymes displayed high kinase activity. This single step purification method provides a convenient tool to follow the status of GSK-3 (protein level, phosphorylation state, kinase activity) under various physiological settings. It also provides a simple and efficient way to purify large amounts of active recombinant or native GSK-3 for screening purposes. (C) 2006 Academic Press.

Original languageEnglish
Pages (from-to)394-404
Number of pages11
JournalProtein Expression and Purification
Issue number3
StatePublished - 2000


  • APC
  • Alzheimer's disease
  • Armadillo
  • Axin
  • Cancer
  • Diabetes
  • GSK-3β
  • Kinase inhibitors
  • Protein kinase
  • Screening
  • WNT
  • β-catenin


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