Unit

UNIT 14.13 Allosteric Activation of Kinases: Design and Application of RapR Kinases

  1. Andrei V. Karginov,
  2. Klaus M. Hahn

Published Online: 1 DEC 2011

DOI: 10.1002/0471143030.cb1413s53

Current Protocols in Cell Biology

Current Protocols in Cell Biology

How to Cite

Karginov, A. V. and Hahn, K. M. 2011. Allosteric Activation of Kinases: Design and Application of RapR Kinases. Current Protocols in Cell Biology. 53:14.13:14.13.1–14.13.16.

Author Information

  1. Department of Pharmacology, University of North Carolina at Chapel Hill, Chapel Hill, North Carolina

Publication History

  1. Published Online: 1 DEC 2011
  2. Published Print: DEC 2011

Abstract

Here we describe a method for the engineered regulation of protein kinases in living cells, the design and application of RapR (rapamycin regulated) kinases. The RapR kinase method enables activation of kinases with high specificity and precise temporal control. Insertion of an engineered allosteric switch, the iFKBP domain, at a structurally conserved position within the kinase catalytic domain makes the modified kinase inactive. Treatment with rapamycin or its non-immunosuppressive analogs triggers interaction with a small FKBP-rapamycin-binding domain (FRB), restoring the activity of the kinase. The reagents used in this method are genetically encoded or membrane permeable, enabling ready application in many systems. Based on the structural similarity of kinase catalytic domains, this method is likely applicable to a wide variety of kinases. Successful regulation has already been demonstrated for three kinases representing both tyrosine and serine/threonine kinase families (p38, FAK, Src). Procedures for designing and testing RapR kinases are discussed. Curr. Protoc. Cell Biol. 53:14.13.1-14.13.16. © 2011 by John Wiley & Sons, Inc.

Keywords:

  • kinase;
  • allosteric;
  • activation;
  • phosphorylation