β-cateninisakeymediatorfortheWntpathway.TheWntpathwayisclassifiedasaβ-catenin-dependentpathwayorβ-catenin-independentpathway.TheWnt/β-catenin-dependentpathwayisresponsIBLeformanycellularregulationssuchasproliferation,differentiationandcell-cellcommunication.HyperactivatedWnt/β-catenin-dependentpathwaytriggersprogressionofmanycancerssuchascolorectalcancers,livercancer,breastcancer,etc.ThetranscriptionoftheoveractiveWnt/β-catenin-dependentpathwayissolelydependentonthenuclearβ-catenin:T-cellfactor(Tcf)binding.Thus,inhibitorstargetingtheβ-catenin/Tcfprotein-proteininteractionappeartobeapromisingtherapeuticstrategyforcancer.Lately,literaturesuggestsactivationoftheβ-catenin/Tcfsignalingbythelipidgrowthfactor,lysophosphatidicacid(LPA),maybeinvolvedinneurogenesisaswell.

Echelon’sβ-catenin/Tcfprotein-proteininteractionassayisafluorescencepolarization(FP)assaydesignedforhighthroughputscreeningina384-wellformatforsmallmoleculeinhibitionortheβ-catenin:Tcfprotein-proteininteraction.Theβ-cateninproteinusedinthisassayisapartialproteinwhichincludesthedomainthatbindstothehumanTcf.AselectedshortpeptidesequenceofthehumanTcfwithTAMRAlabel(Tcf-TAMRA)isusedintheassay.Whenβ-cateninbindstotheTcf-TAMRA,theTAMRAfluorophore’srotationalmovementbecomesslowerduetotheincreaseinmolecularmassandthustheemittedlightremainspolarized(figure1).Therefore,theinhibitionbetweenβ-cateninandTcf-TAMRAbindingcanbemonitoredbythechangeinpolarizationoflight.PKF118-310(Lepourcelet2004)isincludedintheassayasacontrolinhibitor.

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