Improved assay for protein tyrosine phosphatases

G - Physics – 01 – N

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G01N 33/573 (2006.01) C12Q 1/42 (2006.01) C12Q 1/48 (2006.01)

Patent

CA 2435507

The invention relates in part to screening assays for identifying agents that alter the interaction between a protein tyrosine phosphatase (PTP) and its tyrosine phosphorylated polypeptide substrate, using fluorescence energy signals generated by detectably labeled substrates. Assays are provided in certain embodiments, including high throughput screening assays, wherein candidate agents are screened by fluorescence polarization for their ability to influence (i) binding of substrate trapping mutant PTPs to substrates, or (ii) dephosphorylation of tyrosine phosphorylated substrates by PTPs.

La présente invention concerne en partie des tests de criblage permettant d'identifier des agents altérant les interactions entre une protéine tyrosine phosphatase (PTP) et son substrat polypeptidique tyrosine phosphorylé, au moyen de signaux à énergie de fluorescence émis par des substrats marqués de façon à pouvoir être détectés. Des tests sont fournis dans certains modes de réalisation, notamment des tests de criblage à haut rendement, les agents candidats étant criblés par polarisation de fluorescence pour leur capacité à influencer: (I) la liaison des protéine-tyrosine-phosphatases (PTP) mutantes de piégeage aux substrats, ou (II) la déphosphorylation de substrats tyrosine phosphorylés par les PTP.

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