Description | YeastSUMOprotease(alsoknownasUlp),isahighlyactivieandrobustcysteinylproteaseencodedbytheUbl-specificprotease1genefromSaccharomycescerevisiae.ThereareasmanyassixSUMOproteasesinhuman(SENP1–3,SENP5–7).BothhumanandyeastSUMOproteasesrecognizethetertiarystructureoftheubiquitin-like(UBL)protein,i.e.,SUMO-1,SUMO-2andSUMO-3,ratherthanashortstretchofaminoacidsequence,consequently,theseSUMOproteaseswillnotcleavewithinthefusedproteinofinterest.TurboSUMOProteaseisamixtureofcorecatalyticproteasesofbothhumanSENP2andyeastUlp1SUMOproteins,thusSUMO-fusionproteinscanbedigestedbytheTurboSUMOProteaseateither~15°Covernightorat30°Cforafewhours.Afterdigestion,TurboSUMOProteasecanbeeasilyremovedfromthecleavagereactionusingNiagarosebeads |
Application | RemovalofSUMOtagfromfusionproteins |
Source | E.coliover-expressingtheC-terminalSENP2andUlp1catalyticproteasedomainsfromhumanandSaccharomycescerevisiae,respectively. |
Purification | FPLC |
Purity | >95%bySDS-PAGE |
Components | 10U/µlTurboSUMOProteasein50mMHepes,pH7.5,150mMNaCl,0.5mMDTT,50%glycerol |
UnitDefinition | OneunitofSUMOProteaseisdefinedastheamountofenzymeneededtocleave85%of2µgofsubstrateproteinat30°Cor15°Cinonehour |
RecommendedReactionCondition | SUMOProtease:Add1unitto2µgoftargetSUMOfusionprotein Buffer:50mMTris-HCl,pH8.0,0.1%Igepal(NP-40),150mMNaCl,1mMDTT Temperature:30°C(1~3hours),15°C(overnight) OptimalSaltConcentration:imidazole:<150mM;NaCl:<300mM |
Storage | -20°C,orat–80°Cforlong-termstorage |