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Method for the screening of compounds that inhibit the interaction between a proline-rich peptide and a SH3 domain comprising peptide

  • US 20030108975A1
  • Filed: 07/24/2002
  • Published: 06/12/2003
  • Est. Priority Date: 07/30/2001
  • Status: Abandoned Application
First Claim
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1. A method for the screening of a candidate compound for inhibiting the interaction between a proline-rich peptide and a SH3 domain-comprising peptide by Homogeneous Time-Resolved Fluorescence technique, said method comprising the steps of:

  • a) providing a buffer solution containing a proline-rich peptide which is labelled with a first fluorescent marker, wherein said first fluorescent marker is the first partner or the second partner of paired FRET fluorescence markers;

    b) adding the candidate compound to the solution obtained at step a);

    c) adding the SH3 domain-comprising peptide which is directly or indirectly labelled with a second fluorescent marker and wherein said second fluorescent marker is the second partner or the first partner of paired FRET fluorescence markers;

    d) submitting the mixture obtained at step d) to a source of energy at a wavelength corresponding to the excitation wavelength of the first partner of paired FRET fluorescence markers and measuring the fluorescence signal at the emission wavelength of the second partner of paired FRET fluorescence markers;

    e) comparing the fluorescence signal value obtained at step e) with the fluorescence signal value obtained when step b) is omitted to determine if the candidate compound inhibits the interaction between the proline-rich peptide and the SH3 domain-comprising peptide.

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