Purification of the cAMP receptor protein by affinity chromatography

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Summary

8-(6-aminohexyl)-amino-adenosine 3′,5′-monophosphate coupled to agarose has been utilized as affinity chromatography media for separation of the subunits of cAMP-dependent protein kinase and for purification of the regulatory cAMP receptor subunit. Quantitative recovery of highly purified cAMP receptor protein has been achieved using this method. The purified cAMP receptor protein is fully functional in binding cAMP and in reversibly combining with protein kinase catalytic subunits to generate the cAMP-dependent enzyme.

References (22)

  • GillG.N. et al.

    Biochem. Biophys. Res. Commun.

    (1970)
  • KumonA. et al.

    Biochem. Biophys. Res. Commun.

    (1970)
  • ReimannE.M. et al.

    Biochem. Biophys. Res. Commun.

    (1971)
  • ReimannE.M. et al.

    J. Biol. Chem.

    (1971)
  • RubinC.S. et al.

    J. Biol. Chem.

    (1972)
  • YamamuraH. et al.

    J. Biol. Chem.

    (1971)
  • SanbornB.M. et al.

    J. Biol. Chem.

    (1973)
  • ErlichmanJ. et al.

    J. Biol. Chem.

    (1973)
  • CuatrecasasP.

    J. Biol. Chem.

    (1970)
  • WalshD.A. et al.

    J. Biol. Chem.

    (1968)
  • LowryO.H. et al.

    J. Biol. Chem.

    (1951)
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