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In vivo protein screen based on enzyme-assisted chemically induced dimerization ("CID")

  • US 20030138785A1
  • Filed: 02/25/2002
  • Published: 07/24/2003
  • Est. Priority Date: 12/21/2001
  • Status: Abandoned Application
First Claim
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1. A method for identifying which protein from a pool of candidate proteins catalyzes in a cell a bond forming reaction between a first substrate and a second substrate, comprising:

  • (a) providing a dimeric small molecule which comprises a known moiety that binds a known receptor domain covalently linked with a moiety that contains the first substrate;

    (b) introducing the dimeric molecule into a cell which comprises i) a first fusion protein comprising the known receptor domain, ii) a second fusion protein comprising the second substrate, iii) a protein from the pool of candidate proteins, and iv) a reporter gene wherein expression of the reporter gene is conditioned on the proximity of the first fusion protein to the second fusion protein;

    (c) permitting the dimeric molecule to bind to the first fusion protein and to enzymatically form a bond with the second fusion protein so as to activate the expression of the reporter gene;

    (d) selecting which cell expresses the reporter gene; and

    (e) identifying the protein that catalyzes the bond formation reaction in the cell between the first substrate and the second substrate.

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