Method for screening agents modulating a lkb$g(a) protein...

C - Chemistry – Metallurgy – 12 – Q

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C12Q 1/25 (2006.01) C07K 14/47 (2006.01) G01N 33/566 (2006.01)

Patent

CA 2559867

The inventive method for screening agents modulating a lkBa protein ubiquitination by a functional ubiquine ligase protein complex containing .beta.-TrCP protein consists (a) in bringing a testable candidate agent into contact with recombinant yeast cells expressing in the nucleus thereof: (i) a fusion protein comprising I?B? polypeptide and at least a first detectable protein and (ii) a protein comprising a .beta.-TrCP polypeptide, (b) in quantifying said first protein in the yeast cells at the end of at least one predetermined period of time after the contact of the candidate agent with said cells and (c) in comparing the value obtained at the stage (b) with a control value obtained at the stage (a) in the absence of the candidate agent.

Procédé pour le criblage d~agents modulant l~ubiquitination de la protéine I.kappa.B.alpha. par un complexe protéique fonctionnel ubiquitine ligase comprenant la protéine .beta.-TrCP, ledit procédé comprenant les étapes suivantes: (a) mettre en contact un agent candidat à tester avec des cellules de levure recombinantes qui expriment dans leur noyau : (i) une protéine de fusion comprenant le polypeptide I.kappa.B.alpha. et au moins une première protéine détectable ; et (ii) une protéine comprenant le polypeptide .beta.- TrCP ; (b) quantifier ladite première protéine détectable dans les cellules de levure, à la fin d~au moins une période de temps prédéterminée après la mise en contact de l~agent candidat avec lesdites cellules ; (c) comparer la valeur obtenue à l~étape (b) avec une valeur témoin obtenue lorsque l~étape (a) est réalisée en l~absence de l~agent candidat.

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