A rationally‐designed chimeric KDM1A/KDM1B histone demethylase tower domain deletion mutant retaining enzymatic activity

摘要

Atargetwiththerapeuticpotential,lysine‐specificdemethylase1A(KDM1A)isaregulatorofgeneexpressionwhosetowerdomainisaprotein–proteininteractionmotif.ThisdomainfacilitatestheinteractionofKDM1Awithcoregulatorsandmultiproteincomplexesthatdirectitsactivitytonucleosomes.Wedescribethedesignandcharacterizationofachimeric‘towerless’KDM1A,termednΔ150KDM1AΔTowerKDM1Bchimera(chKDM1AΔTower),whichincorporatesaregionfromtheparaloglysine‐specificdemethylase1B(KDM1B).ThischimeracopurifieswithFADanddisplaysdemethylaseactivity,butfailstobindthepartnerproteincorepressoroftheRE1‐silencingtranscriptionfactor(CoREST).WeconcludethatKDM1Acatalysiscanbedecoupledfromtower‐dependentinteractions,lendingchKDM1AΔTowerusefulfordissectingmolecularcontributionstoKDM1Afunction...