MECHANISM BASED CROSS-LINKERS FOR IDENTIFYING KINASE-SUBSTRATE INTERACTIONS

BACKGROUND:

Protein kinases are enzymes that act at critical control points in many cellular processes, including the pathways responsible for regulating cell growth and differentiation. This central role makes protein kinases a very attractive target for drug design and treatment of diseases. Understanding which proteins interact in a given pathway is an important step towards modulating those pathways for therapeutic benefit. Standard methods for identifying protein-protein interactions such as 2-hybrid screens and pull-down assays are not generally effective in identifying kinase-substrate interactions due to their transient, low affinity binding. Molecular tools that capture kinase-substrate interactions are needed to map the multiple protein-protein interactions that control many cellular functions.

DESCRIPTION:

Researchers at the University of California, San Francisco have synthesized and validated a small molecule mechanism-based cross-linker capable of trapping kinase-substrate interactions. This new research tool has been designed to cross-link any serine/threonine kinase to its biological substrate through binding to the conserved kinase ATP-binding site and reacting with an invariant active-site lysine. Specificity for the substrate is obtained by replacing the normally phosphorylated substrate serine/threonine with a cysteine residue. This system allows validation of putative kinase-substrate binding as well as identification of unknown kinase-substrate interactions. The cross-linker has been tested under different solution conditions and with Akt1, p38 MAP kinase, PKA, and casein kinase II, showing the system to be robust and capable of cross-linking a wide range of kinases. Researchers are also testing the cross-linker with tyrosine kinases to further expand the utility of this tool.

PUBLICATIONS AND PATENTS

• A mechanism-based cross-linker for the identification of kinase-substrate pairs, Maly et al., J. Am. Chem. Soc.; 2004; 126(30) pp 9160 – 9161.


• Mechanism-Based Crosslinkers, International application no. PCT/US2005/026359, Publication no. WO/2006/012624.

BENEFITS AND APPLICATIONS FOR THIS TECHNOLOGY:

• Small molecule cross-linker for trapping kinase-substrate binding


• Capable of validating putative partners or identifying new kinase-substrate interactions


• Validated with multiple kinases and under different solution conditions
  

If you would like to receive further information about this technology and potential licensing opportunities, please contact:

Ellen S. Kats, Ph.D.
Licensing Associate
(415) 514-8210 phone
(415) 348-1579 fax
Ellen.Kats@ucsf.edu

Reference: OTM Case #SF2004-068