TMR-ShK peptidetoxinisasyntheticderivativeofthewell-known ShKtoxin(Stichodactylahelianthusneurotoxin) isolatedfromthevenomoftheCarribeanseaanemone Stoichactishelianthus.Wild-type ShK blockspotentlyKv1.3(KCNA3),Kv1.1(KCNA1),Kv1.4(KCNA4)andKv1.6(KCNA6)withaKdvalueof11pM,16pM,312pMand165pM,respectively.ThefluorescentTMR-labeledShKblocksKv1.3andKv1.1withKdvaluesof52pMand397pM,respectively. TMR-ShK canbeusedtoidentifylymphocytesexpressingKv1.3channelssuchaseffectivememoryTcells(TEM)inthecaseofautoimmunediseases(type1diabetes,mellitusorrheumatoidarthritis)whichoverexpressKv1.3channels.
Productcode:SAT001-00100.Categories:Kv1.3channel,Potassiumchannels.Tags:Kv1.3,TRAM-34.
AAsequence: TMR-AEEAc-Arg-Ser-Cys3-Ile-Asp-Thr-Ile-Pro-Lys-Ser-Arg-Cys12-Thr-Ala-Phe-Gln-Cys17-Lys-His-Ser-Met-Lys-Tyr-Arg-Leu-Ser-Phe-Cys28-Arg-Lys-Thr-Cys32-Gly-Thr-Cys35-OH
Disulfidebonds: Cys3-Cys35,Cys12-Cys28 andCys17-Cys32
Length(aa): 35
Formula: C200H310N57O55S7
MolecularWeight: 4611.7Da
Appearance:redlyophilizedsolid
Solubility: waterorsalinebuffer
CASnumber: NA
Source: Synthetic
Purityrate: >97%
TMRfluorescentdye (5(6)-TAMRA): λex543nM, λem572nM
Tlymphocyteswithunusuallyhighexpressionofthevoltage-gatedKv1.3channel(Kv1.3(high)cells)havebeenimplicatedinthepathogenesisofexperimentalautoimmuneencephalomyelitis,ananimalmodelformultiplesclerosis.WehavedevelopedafluoresceinatedanalogofShK(ShK-F6CA),themostpotentknowninhibitorofKv1.3,fordetectionofKv1.3(high)cellsbyflowcytometry.ShK-F6CAblockedKv1.3atpicomolarconcentrationswithaHillcoefficientof1andexhibited>80-foldspecificityforKv1.3overKv1.1andotherK(V)channels.Inflowcytometryexperiments,ShK-F6CAspecificallystainedKv1.3-expressingcellswithadetectionlimitofapproximately600channelspercell.RatandhumanTcellsthathadbeenrepeatedlystimulated7-10timeswithantigenwerere
ADIlydistinguishedonthebasisoftheirhighlevelsofKv1.3channels(>600channels/cell)andShK-F6CAstainingfromrestingTcellsorcellsthathadundergone1-3roundsofactivation.FunctionalKv1.3expressionlevelsincreasedsubstantiallyinamyelin-specificratTcelllinefollowingmyelinantigenstimulation,peakingat15-20handthendecliningtobaselineoverthenext7days,inparallelwiththeacquisitionandlossofencephalitogenicity.Bothcalcium-andproteinkinaseC-dependentpathwayswererequiredfortheantigen-inducedKv1.3up-regulation.ShK-F6CAmightbeusefulforrapidandquantitativedetectionofKv1.3(high)expressingcellsinnormalanddiseasedtissues,andtovisualizethedistributionoffunctionalchannelsinintactcells.BeetonC.,etal.(2003)ANovelFluorescentToxintoDetectandInvestigateKv1.3ChannelUp-regulationinChronicallyActivatedTLymphocytes.JBC.PMID:12511563
