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Interactions between RaF proto-oncogenes and CDC25 phosphatases, and uses related thereto

  • US 6,037,136 A
  • Filed: 10/24/1994
  • Issued: 03/14/2000
  • Est. Priority Date: 10/24/1994
  • Status: Expired due to Fees
First Claim
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1. A method for identifying a compound which is an inhibitor of ras-mediated activation of a CDC25 phosphatase, comprising the steps of:

  • a) generating a reconstituted protein combination including;

    1) a test agent to be assessed;

    2) a purified protein preparation of a mammalian CDC25 phosphatase,3) a purified protein preparation of a kinase from a ras-mediated signal transduction cascade, which kinase phosphorylates the CDC25 phosphatase; and

    4) a substrate of the CDC25 phosphatase;

    b) maintaining the combination under conditions appropriate for the kinase to phosphorylate the CDC25 phosphatase and for an activated CDC25 phosphatase to convert the substrate to product; and

    c) measuring the conversion of the substrate to product,wherein a decrease in the conversion of substrate to product in the combination, relative to a control comprising the CDC25 phosphatase, the kinase, and the substrate and lacking the test agent, indicates that the test compound is an inhibitor of ras-mediated activation of the CDC25 phosphatase.

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