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Engineered allosteric activation of kinases in living cells.

Studies of cellular and tissue dynamics benefit greatly from tools that can control protein activity with specificity and precise timing in living systems. Here we describe an approach to confer allosteric regulation specifically on the catalytic activity of protein kinases. A highly conserved portion of the kinase catalytic domain is modified with a small protein insert that inactivates catalytic activity but does not affect other protein functions (Fig. 1a). Catalytic activity is restored by addition of rapamycin or non-immunosuppresive rapamycin analogs. Molecular modeling and mutagenesis indicate that the protein insert reduces activity by increasing the flexibility of the catalytic domain. Drug binding restores activity by increasing rigidity. We demonstrate the approach by specifically activating focal adhesion kinase (FAK) within minutes in living cells and show that FAK is involved in the regulation of membrane dynamics. Successful regulation of Src and p38 by insertion of the rapamycin-responsive element at the same conserved site used in FAK suggests that our strategy will be applicable to other kinases.

Pubmed ID: 20581846


  • Karginov AV
  • Ding F
  • Kota P
  • Dokholyan NV
  • Hahn KM


Nature biotechnology

Publication Data

July 12, 2010

Associated Grants

  • Agency: NIGMS NIH HHS, Id: GM057464
  • Agency: NIGMS NIH HHS, Id: GM080742
  • Agency: NIGMS NIH HHS, Id: GM080742- 03S1
  • Agency: NIGMS NIH HHS, Id: GM64346
  • Agency: NIGMS NIH HHS, Id: R01 GM057464
  • Agency: NIGMS NIH HHS, Id: R01 GM057464-10
  • Agency: NIGMS NIH HHS, Id: U54 GM064346
  • Agency: NIGMS NIH HHS, Id: U54 GM064346-099026

Mesh Terms

  • Allosteric Regulation
  • Catalytic Domain
  • Models, Molecular
  • Mutagenesis
  • Phosphorylation
  • Protein Kinases