Basedonproteindomainsknowntopossessanaffinityforubiquitin,TandemUbiquitinBindingEntities(TUBEs)havebeendevelopedfortheisolationandidentificationofubiquitylatedproteins.TUBEsdisplayuptoa1000-foldincreaseinaffinityforpoly-ubiquitinmoietiesoverthesingleubiquitinbindingassociateddomain(UBA).Inaddition,TUBEsdisplayaprotectiveeffectonpolyubiquitinatedproteins,allowingfordetectionatrelativelylowabundance.Thesepropertieseffectively“capture”proteinsintheirpolyubiquitylatedstate.
Theubiquitylationstateofcertaincellularproteinscanbedirectlyrelatedtovariousdiseasesincludingneurodegeneration,inflammation,andcancer.However,ubiquitylatedproteinsareintrinsicallyunstable,whichprecludesthecharacterizationoftheubiquitinylationstateinmanycases.TUBEswillfillthisvoidinubiquitinresearch,allowingtheidentificationofubiquitin-modifiedproteinsthatcannotbedetectedwithcurrenttechnologies.TUBEscanbeusedtoisolate,enrich,andidentifyubiquitylatedproteinsfromcells,tissuesandorgans.
TUBE2isbasedonUBA1fromhumanRAD23A.TheaffinityofTUBE2forK63linkedtetra-ubiquitinandK48tetra-ubiquitinlinkedchainsisequivalent.
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Info
| Molecularweight | 38.5kD |
| AffinityTag | His6 |
| Affinity | His6-TUBE2displaysequivalentaffinitytowardsK48andK63-linkedtetra-ubiquitin. |
Applications
- Pulldownofpoly-ubiquitylatedproteinsfromcell andtissuelysates
- Protectionofpoly-ubiquitylatedproteinsfrombothdeubiquitylationanddegradationbytheproteasome
Benefits
- Upto1000-foldhigheraffinitythanthesingleUBAdomain
- Avoidoverexpressionofaffinity-taggedubiquitinsforpulldowns
- Protectpoly-ubiquitinchainswithoutinhibitorsspecifictotheproteasomeordeubiquitylaseactivity
- Replaceubiquitinspecificantibodiesforenrichmentofpoly-ubiquitylatedproteins
Documents
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那糖酵解中依赖NAD的酶不是要在线粒体内反应?
蛋白酶分布广泛,主要存在于人和动物消化道中,而消化道通过口与肛门与外界相通,属于外环境或外界环境
(溶菌酶、凝血酶原等)分布在内环境
(消化)酶分布在消化道

