Determination of the Kinetics and Thermodynamics of Ligand Binding to a Specific Inactive Conformation in Protein Kinases

  • Sanjay B. HariEmail author
  • Pratistha Ranjitkar
  • Dustin J. Maly
Part of the Methods in Molecular Biology book series (MIMB, volume 928)


Recent interest in inactive kinase conformations has generated the need to develop new biochemical tools to study them. Here, we describe the use of a fluorescent probe that selectively and potently binds to a specific inactive conformation of protein kinases. This allows for the thermodynamics and kinetics of ligand binding to be determined.

Key words

Kinase Inactive conformation DFG-out Activation loop Fluorescent probe Protein ligand Dissociation constant 



This work was supported by the National Institute of General Medical Science (R01GM086858).


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Copyright information

© Springer Science+Business Media New York 2012

Authors and Affiliations

  • Sanjay B. Hari
    • 1
    Email author
  • Pratistha Ranjitkar
    • 1
  • Dustin J. Maly
    • 1
  1. 1.Department of ChemistryUniversity of WashingtonSeattleUSA

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