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Kinase and phosphatase assays

  • US 7,727,752 B2
  • Filed: 07/14/2006
  • Issued: 06/01/2010
  • Est. Priority Date: 07/29/2003
  • Status: Active Grant
First Claim
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1. A method for identifying a modulator of activity of a kinase, comprising:

  • a) forming a kinase mixture comprising a protein kinase, a composition, and a test compound;

    b) contacting the kinase mixture with a protease to form a protease mixture;

    c) forming a control mixture comprising the protein kinase, the composition, and the protease; and

    d) comparing a measurable property in the protease mixture to the measurable property in the control mixture,wherein the test compound is identified as a modulator of activity of the kinase if the measurable property in the protease mixture is different from the measurable property in the control mixture, andwherein the composition comprises;

    (i) a peptide having a length from five to fifty amino acids, the peptide comprising a motif, wherein the motif is;

    TX1YVA, where X1 can be G, A, or E (SEQ ID NO;

         10);

    (ii) a first detectable moiety, wherein the first detectable moiety is associated with the peptide; and

    (iii) a protease cleavage site, wherein the protease cleavage site is located in a position relative to the motif such that enzymatic modification of the motif alters the proteolytic cleavage of the peptide;

    wherein the protein kinase is selected from the group consisting of CSF1R,FLT 3, and c-Kit.

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