(Dap22)-ShK(ShKDap22)peptideisasyntheticderivativeofthewell-knownShKtoxin#08SHK001(Stichodactylahelianthusneurotoxin)isolatedfromthevenomoftheCarribeanseaanemoneStoichactishelianthus.Wild-typeShKblockspotentlyKv1.3(KCNA3),Kv1.1(KCNA1),Kv1.4(KCNA4),andKv1.6(KCNA6)respectivelywithaKdof11pM,16pM,312pMand165pM.In(Dap22)-ShK,lysine22hasbeenreplacedbyadiaminopropionicacid(Dap)residuethatgreatlyimprovestheselectivityofthepeptideforthevoltage-gatedpotassiumchannelKv1.3(IC50around23pM)againstKv1.1(1.8nM),Kv1.4(37nM)andKv1.6(10nM)channels.Thehighselectivityof(Dap22)-ShkisachievedthankstothestrongbindingbetweentheDapandHis404/Asp386residuesofKv1.3channel.(Dap22)-ShKinhibitsTcellproliferationinducedbyanti-CD3atsubnanomolarconcentrations.
Description:
AAsequence:Arg-Ser-Cys3-Ile-Asp-Thr-Ile-Pro-Lys-Ser-Arg-Cys12-Thr-Ala-Phe-Gln-Cys17-Lys-His-Ser-Met-Dap-Tyr-Arg-Leu-Ser-Phe-Cys28-Arg-Lys-Thr-Cys32-Gly-Thr-Cys35-OH
Disulfidebonds:Cys3-Cys35,Cys12-Cys28andCys17-Cys32
Length(aa):35
Formula:C166H268N54O48S7
MolecularWeight:4012.8Da
Appearance:Whitelyophilizedsolid
Solubility:waterandsalinebuffer
CASnumber:220384-25-8
Source:Synthetic
Purityrate:>97%
Reference:
HypoxiamodulatesearlyeventsinTcellreceptor-mediatedactivationinhumanTlymphocytesviaKv1.3channels
Tlymphocytesareexposedtohypoxiaduringtheirdevelopmentandwhentheymigratetohypoxicpathologicalsites.AlthoughithasbeenshownthathypoxiainhibitsKv1.3channelsandproliferationinhumanTcells,themechanismsbywhichhypoxiaregulatesTcellactivationarenotfullyunderstood.HereinwetestthehypothesisthathypoxicinhibitionofKv1.3channelsinducesmembranedepolarization,thusmodulatingtheincreaseincytoplasmicCa2+thatoccursduringactivation.HypoxiacausesmembranedepolarizationinhumanCD3+Tcells,asmeasuredbyfluorescence-activatedcellsorting(FACS)withthevoltage-sensitivedyeDiBAC4(3).SimilardepolarizationisproducedbytheselectiveKv1.3channelblockersShK-Dap22andmargatoxin.FurThermore,pre-exposuretosuchblockerspreventsanyfurtherdepolarizationbyhypoxia.SincemembranedepolarizationisunfavourabletotheinfluxofCa2+throughtheCRACchannels(necessarytodrivemanyeventsinTcellactivationsuchascytokineproductionandproliferation),theeffectofhypoxiaonTcellreceptor-mediatedincreaseincytoplasmicCa2+wasdeterminedusingfura-2.HypoxiadepressestheincreaseinCa2+inducedbyanti-CD3/CD28antibodiesinapproximately50%oflymphocytes.Intheremainingcells,hypoxiaeitherdidnotelicitanychangeorproducedasmallincreaseincytoplasmicCa2+.Similareffectswereobservedinrestingandpre-activatedCD3+cellsandweremimickedbyShK-Dap22.TheseeffectsappeartobemediatedsolelybyKv1.3channels,aswefindnoinfluenceofhypoxiaonIKCa1andCRACchannels.OurfindingsindicatethathypoxiamodulatesCa2+homeostasisinTcellsviaKv1.3channelinhibitionandmembranedepolarization.
RobbinsJR.,etal.(2005)HypoxiamodulatesearlyeventsinTcellreceptor-mediatedactivationinhumanTlymphocytesviaKv1.3channels.JPhysiol.PMID: 15677684
PotassiumchannelblockadebytheseaanemonetoxinShKforthetreatmentofmultiplesclerosisandotherautoimmunediseases
Expressionofthetwolymphocytepotassiumchannels,thevoltage-gatedchannelKv1.3andthecalciumactivatedchannelIKCa1,changesduringdifferentiationofhumanTcells.WhileIKCa1isthefunctionallydominantchannelinnaiveand“early”memoryTcells,Kv1.3iscrucialfortheactivationofterminallydifferentiatedeffectormemory(TEM)Tcells.BecauseoftheinvolvementofTEMcellsinautoimmuneprocesses,Kv1.3isregardedasapromisingtargetforthetreatmentofT-cellmediatedautoimmunediseasessuchasmultiplesclerosisandthepreventionofchronictransplantrejection.ShK,a35-residuepolypeptidetoxinfromtheseaanemone,Stichodactylahelianthus,blocksKv1.3atlowpicomolarconcentrations.ShKadoptsacentralhelix-kink-helixfold,andalanine-scanningandothermutagenesisstudieshavedefineditschannel-bindingsurface.ModelshavebeendevelopedofhowthistoxineffectsK+-channelblockadeandhowitsdockingconfigurationmightdifferinShK-Dap22,whichcontainsasinglesidechainsubstitutionthatconfersspecificityforKv1.3blockade.ShK,ShK-Dap22andtheKv1.3blockingscorpiontoxinkaliotoxinhavebeenshowntopreventandtreatexperimentalautoimmuneencephalomyelitisinrats,amodelformultiplesclerosis.AfluoresceinatedanalogofShK,ShK-F6CA,hasbeendeveloped,whichallowsthedetectionofactivatedTEMcellsinhumanandanimalbloodsamplesbyflowcytometryandthevisualizationofKv1.3channeldistributioninlivingcells.ShKanditsanalogsarecurrentlyundergoingfurtherevaluationasleadsinthedevelopmentofnewbiopharmaceuticalsforthetreatmentofmultiplesclerosisandotherT-cellmediatedautoimmunedisorders.
NortonRS.,etal.(2004)PotassiumchannelblockadebytheseaanemonetoxinShKforthetreatmentofmultiplesclerosisandotherautoimmunediseases.CurrMedChem. PMID: 15578998
SubstitutionofasingleresidueinStichodactylahelianthuspeptide,ShK-Dap22,revealsanovelpharmacologicalprofile
ShK,apeptideisolatedfromStichodactylahelianthusvenom,blocksthevoltage-gatedpotassiumchannels,K(v)1.1andK(v)1.3,withsimilarhighaffinity.ShK-Dap(22),asyntheticderivativeinwhichadiaminopropionicacidresiduehasbeensubstitutedatpositionLys(22),hasbeenreportedtobeaselectiveK(v)1.3inhibitorandtoblockthischannelwithequivalentpotencyasShK[Kalmanetal.(1998)J.Biol.Chem.273,32697-32707].Inthisstudy,alargebodyofevidenceispresentedwhichindicatesthatthepotenciesofwild-typeShKpeptideforbothK(v)1.3andK(v)1.1channelshavebeenpreviouslyunderestimated.Therefore,theaffinityofShK-Dap(22)forbothchannelsappearstobeca.10(2)-10(4)-foldweakerthanShK.ShK-Dap(22)doesdisplayca.20-foldselectivityforhumanK(v)1.3vsK(v)1.1whenmeasuredbythewhole-cellvoltageclampmethodbutnotinequilibriumbindingassays.ShK-Dap(22)haslowaffinityforK(v)1.2channels,butheteromultimericK(v)1.1-K(v)1.2channelsformareceptorwithca.200-foldhigheraffinityforShK-Dap(22)thanK(v)1.1homomultimers.Infact,K(v)1.1-K(v)1.2channelsbindShK-Dap(22)withonlyca.10-foldlesspotencythanShKandrevealanovelpharmacologynotpredictedfromthehomomultimersofK(v)1.1orK(v)1.2.TheconcentrationsofShK-Dap(22)neededtoinhibithumanTcellactivationwereca.10(3)-foldhigherthanthoseofShK,ingoodcorrelationwiththerelativeaffinitiesofthesepeptidesforinhibitingK(v)1.3channels.Allofthesedata,takentogether,suggestthatShK-Dap(22)willnothavethesameinvivoimmunosuppressantefficacyofotherK(v)1.3blockers,suchasmargatoxinorShK.Moreover,ShK-Dap(22)mayhaveundesiredsideeffectsduetoitsinteractionwithheteromultimericK(v)1.1-K(v)1.2channels,suchasthosepresentinbrainand/orperipheraltissues.
MiddletonRE.,etal.(2003)SubstitutionofasingleresidueinStichodactylahelianthuspeptide,ShK-Dap22,revealsanovelpharmacologicalprofile.Biochemistry. PMID: 14622016
MutatingacriticallysineinShKtoxinaltersitsbindingconfigurationinthepore-vestibuleregionofthevoltage-gatedpotassiumchannel,Kv1.3
Thevoltage-gatedpotassiumchannelinTlymphocytes,Kv1.3,animportanttargetforimmunosuppressants,isblockedbypicomolarconcentrationsofthepolypeptideShKtoxinanditsanalogueShK-Dap22.ShK-Dap22showsincreasedselectivityforKv1.3,andourgoalwastodeterminethemolecularbasisforthisselectivitybyprobingtheinteractionsofShKandShK-Dap22withtheporeandvestibuleofKv1.3.Thefreeenergiesofinteractionsbetweentoxinandchannelresiduesweremeasuredusingmutantcycleanalyses.Thesedata,interpretedasapproximatedistancerestraints,guidedmoleculardynamicssimulationsinwhichthetoxinsweredockedwithamodelofKv1.3basedonthecrystalstructureofthebacterialK(+)-channelKcsA.Despitethesimilartertiarystructuresofthetwoligands,themutantcycledataimplythattheymakedifferentcontactswithKv1.3,andtheycanbedockedwiththechannelinconfigurationsthatareconsistentwiththemutantcycledataforeachtoxinbutquitedistinctfromoneanother.ShKbindstoKv1.3withLys22occupyingthenegativelychargedporeofthechannel,whereastheequivalentresidueinShK-Dap22interactswithresiduesfurtheroutinthevestibule,producingasignificantchangeintoxinorientation.TheincreasedselectivityofShK-Dap22isachievedbystronginteractionsofDap22withHis404andAsp386onKv1.3,withonlyweakinteractionsbetweenthechannelporeandthetoxin.PotentandspecificblockadeofKv1.3apparentlyoccurswithoutinsertionofapositivelychargedresidueintothechannelpore.Moreover,thefindingthatasingleresiduesubstitutionaltersthebindingconfigurationemphasizestheneedtoobtainconsistentdatafrommultiplemutantcycleexperimentsinattemptstodefineproteininteractionsurfacesusingthesedata.
LaniganMD.,etal.(2002)MutatingacriticallysineinShKtoxinaltersitsbindingconfigurationinthepore-vestibuleregionofthevoltage-gatedpotassiumchannel,Kv1.3.Biochemistry. PMID: 12356296
SelectiveblockadeofTlymphocyteK(+)channelsamelioratesexperimentalautoimmuneencephalomyelitis,amodelformultiplesclerosis
Adoptivetransferexperimentalautoimmuneencephalomyelitis(AT-EAE),adiseaseresemblingmultiplesclerosis,isinducedinratsbymyelinbasicprotein(MBP)-activatedCD4(+)Tlymphocytes.Bypatch-clampanalysis,encephalitogenicratTcellsstimulatedrepeatedlyinvitroexpressedauniquechannelphenotype(“chronicallyactivated”)withlargenumbersofKv1.3voltage-gatedchannels(approximately1500percell)andsmallnumbersofIKCa1Ca(2+)-activatedK(+)channels(approximately50-120percell).Incontrast,restingTcellsdisplayed0-10Kv1.3and10-20IKCa1channelspercell(“quiescent”phenotype),whereasTcellsstimulatedonceortwiceexpressedapproximately200Kv1.3andapproximately350IKCa1channelspercell(“acutelyactivated”phenotype).Consistentwiththeirchannelphenotype,[(3)H]thymidineincorporationbyMBP-stimulatedchronicallyactivatedTcellswassuppressedbythepeptideShK,ablockerofKv1.3andIKCa1,andbyananalog(ShK-Dap(22))engineeredtobehighlyspecificforKv1.3,butnotbyaselectiveIKCa1blocker(TRAM-34).ThecombinationofShK-Dap(22)andTRAM-34enhancedthesuppressionofMBP-stimulatedTcellproliferation.Basedontheseinvitroresults,weassessedtheefficacyofK(+)channelblockersinAT-EAE.SpecificandsimultaneousblockadeoftheTcellchannelsbyShKorbyacombinationofShK-Dap(22)plusTRAM-34preventedlethalAT-EAE.BlockadeofKv1.3alonewithShK-Dap(22),butnotofIKCa1withTRAM-34,wasalsoeffective.WhenadmiNISTeredaftertheonsetofsymptoms,ShKorthecombinationofShK-Dap(22)plusTRAM-34greatlyamelioratedtheclinicalcourseofbothmoderateandsevereAT-EAE.WeconcludethatselectivetargetingofKv1.3,aloneorwithIKCa1,mayprovideaneffectivenewmodeoftherapyformultiplesclerosis.
BeetonC.,etal.(2001)SelectiveblockadeofTlymphocyteK(+)channelsamelioratesexperimentalautoimmuneencephalomyelitis,amodelformultiplesclerosis.ProcNatlAcadSciUSA. PMID: 11717451
ShK-Dap22,apotentKv1.3-specificimmunosuppressivepolypeptide
Thevoltage-gatedpotassiumchannelinTlymphocytes,Kv1.3,isanimportantmoleculartargetforimmunosuppressiveagents.Astructurallydefinedpolypeptide,ShK,fromtheseaanemoneStichodactylahelianthusinhibitedKv1.3potentlyandalsoblockedKv1.1,Kv1.4,andKv1.6atsubnanomolarconcentrations.UsingmutantcycleanalysisinconjunctionwithcomplementarymutagenesisofShKandKv1.3,andutilizingthestructureofShK,wedeterminedalikelydockingconfigurationforthispeptideinthechannel.Baseduponthistopologicalinformation,wereplacedthecriticalLys22inShKwiththepositivelycharged,non-naturalaminoaciddiaminopropionicacid(ShK-Dap22)andgeneratedahighlyselectiveandpotentblockeroftheT-lymphocytechannel.ShK-Dap22,atsubnanomolarconcentrations,suppressedanti-CD3inducedhumanT-lymphocyte[3H]thymidineincorporationinvitro.Toxicitywiththismutantpeptidewaslowinarodentmodel,withamedianparalyticdoseofapproximately200mg/kgbodyweightfollowingintravenousadministration.TheoverallstructureofShK-Dap22insolution,asdeterminedfromNMRdata,issimilartothatofnativeShKtoxin,buttherearesomedifferencesintheresiduesinvolvedinpotassiumchannelbinding.Basedontheseresults,weproposethatShK-Dap22orastructuralanaloguemayhaveuseasanimmunosuppressantforthepreventionofgraftrejectionandforthetreatmentofautoimmunediseases.
KalmanK,etal.(1998)ShK-Dap22,apotentKv1.3-specificimmunosuppressivepolypeptide.JBiolChem. PMID: 9830012
Smartox Biotechnolgy的多肽毒素产品如下：
1. 作用于钠离子通道(Sodium channel)的毒素
Toxin name | Catalog # | Target |
Phrixotoxin-3 | 13PHX003 | Selective blocker of Nav1.2 |
µ-conotoxin GIIIB | CON020 | Selective blocker of Nav1.4 |
µ-conotoxin CnIIIC | CON021 | Selective blocker of Nav1.4 |
μ-conotoxin PIIIA | 08CON006 | Selective blocker of Nav1.4 |
Jingzhaotoxin-III | 12JZH003 | Selective blocker of Nav1.5 |
ProTx-II | 07PTX002 | Selective blocker of Nav1.7 |
ProTx-II Biotin | 12PTB002 | Selective blocker of Nav1.7 |
ProTx-I | 12PTX001 | Blocker of Nav1.8, Nav1.2, Nav1.5, Nav1.7 |
Huwentoxin-I | 07HWT001 | Blocker of TTX-S |
Huwentoxin-IV | 08HWT002 | Blocker of TTX-S |
Hainantoxin-III | 13HTX003 | Blocker of TTX-S |
Hainantoxin-IV | 12HTX001 | Blocker of TTX-S |
GsAF-I | 12GSF001 | Blocker of TTX-S |
GsAF-II | 12GSF002 | Blocker of TTX-S |
2. 作用于钾离子通道(Potassium channel)的毒素
Toxin name | Catalog # | Target |
KCa channels | ||
Apamin 蜜蜂神经毒素 | 08APA001 | SK1, SK2, SK3 |
Charybdotoxin 蝎毒素 | 11CHA001 | KCa1.1, KCa3.1 - Kv1.2, Kv1.3, Kv1.6 |
Iberiotoxin | 12IBX001 | KCa1.1 |
Leiurotoxin 1 (Scyllatoxin) | 10LEI001 | SK1, SK2, SK3 |
Tamapin | 10TAM001 | SK1, SK2, SK3 |
Kaliotoxin-1 | 08KTX002 | BK, Kv1.1, Kv1.2, Kv1.3 |
Kv channels | ||
ShK | 08SHK001 | Kv1.3, Kv1.1, Kv1.4, Kv1.6 |
TMR-ShK | SAT001 | Kv1.3, Kv1.1 |
Margatoxin | 08MAG001 | Kv1.3 |
(Dap22)-ShK | 13SHD001 | Kv1.3 |
ADWX-1 | 13ADW001 | Kv1.3 |
HsTx1 | 08NEU001 | Kv1.3, Kv1.2 |
Agitoxin-2 | 13AGI002 | Kv1.3, Kv1.1 |
Maurotoxin | 08MAR001 | Kv1.2, KCa3.1 |
Guangxitoxin 1E | 11GUA002 | Kv2.1, Kv2.2 |
Stromatoxin 1 NEW | SCT01 | Kv2.1, Kv2.2 |
Kaliotoxin-1 | 08KTX002 | BK, Kv1.1, Kv1.2, Kv1.3 |
Charybdotoxin | 11CHA001 | KCa1.1, KCa3.1 - Kv1.2, Kv1.3, Kv1.6 |
Phrixotoxin-2 | PHX002 | Kv4.2, Kv4.3 |
AmmTx3 NEW | AMX001 | A-type potassium channels |
Inwardly rectifying potassium channels | ||
TertiapinQ | 08TER001 | Kir1.1, Kir3.1/3.4, Kir3.1/3.2-KCa1.1 |
hERG/Kv11.1 | ||
BeKm-1 | 13BEK001 | ERG1 |
3. 作用于钙离子通道(Calcium channel)的毒素
Toxin name | Catalog # | Target |
High voltage-gated Ca2+ channels | ||
ω-agatoxin IVA | 11AGA001 | P/Qtype |
ω-Conotoxin MVIIC | 08CON002 | P/Qtype, N-type |
ω-Conotoxin MVIIA | 08CON001 | N-type |
ω-Conotoxin GVIA | 08CON003 | N-type |
ω-Conotoxin SO3 | 08CON013 | N-type |
Huwentoxin I | 07HWT001 | N-type |
ProTx-II | 07PTX002 | T-type, L-type |
Intermediate voltage-gated Ca2+ channels | ||
SNX482 | 08SNX002 | R-type |
Low voltage-gated Ca2+ channels | ||
ProTx-I | 12PTX001 | T-type |
ProTx-II | 07PTX002 | T-type, L-type |
Ryanodine receptors | ||
Maurocalcine | 07PAU001 | Ryr1 |
4. 作用于氯离子通道(Chloride channel)的毒素
Toxin name | Catalog # | Target |
Chlorotoxin | 08CHL001 | Blocker of small conductance Cl- channels |
GaTx1 | 13GTX001 | Selective blocker of CFTR channel |
GaTx2 | 10GTX002 | Selective blocker of ClC-2 channel |
5. 作用于乙酰胆碱受体(Acetylcholine receptor)的毒素
Toxin name | Catalog # | Target |
α-conotoxin PeIA | 13CON017 | α9α10, α3β2 subunits |
α-Conotoxin PrXA | 13CON016 | α1/β1/ε/δ, α1/β1/γ/δ subunits |
Waglerin-1 | 12WAG001 | MusclenAChR |
α-conotoxin MI | 08CON012 | α1/δsubunits |
α-conotoxin GI | 08CON005 | α/δsite |
α-conotoxin IMI | 08CON011 | α7 homomeric nAChR |
α-conotoxin GID | CON019 | Blocker of α3β2, α7 and α4β2 nAChRs |
6. 含N-甲基-D-天冬氨酸NR2B
(NMDA, NR2B containing N-methyl-D-aspartate)
Conantokin-G
选择性、特异性抑制含NR2B的NMDAR。Conantokin-G能剂量依赖性抑制Ca2+内流，抑制NMDA诱导的兴奋性中毒效应。研究表明，在小鼠皮层神经元，Conantokin-G阻滞NMDA引发的电流信号的IC50值为480 nM。
7. 作用于酸敏感离子通道(ASIC channel, Acid-Sensing Ion Channel)的毒素
Toxin name | Catalog # | Target |
APETx2 | 07APE002 | Selective blocker of ASIC3 |
Psalmotoxin1/PcTx1 | 13PCT001 | Selective blocker of ASIC1a |
Ugr9-1 | 13UGR001 | Blocker of ASIC3 |
8. 作用于瞬时受体电位(TRP channel, transient receptor potential)的毒素
Toxin name | Catalog # | Target |
GsMTx4 | 08GSM001 | TRPC, TRPA |
Vanillotoxin3 | 10VAN003 | Activator of TRPV1 |
ProTx-I | 12PTX001 | Antagonist of TRPA1 |
9. 作用于嘌呤能通道(Purinergic channel)的毒素
Purotoxin-1
选择性抑制P2X3受体。100 nM Purotoxin-1 (PT-1)选择性抑制P2X3受体通道，在大鼠DRG神经元上，使用膜片钳实验表明：PT-1对电压门控通道和TRPV1均无抑制效应。10 µM ATP和100 µM α,β Methylene-ATP浓度下Purotoxin-1对P2X3受体有选择性作用，在该ATP浓度下Purotoxin-1对P2X2和杂化二聚体P2X2/3并无激动作用。Purotoxin-1对疼痛的潜在治疗作用。
10. 作用于其它膜受体通道(Others)的毒素
Smartox Biotechnology还提供其他类型的膜受体抑制剂：
Toxin name | Catalog # | Target |
Morphiceptin | 011CAS001 | Agonist of µ-opoid receptors |
Lys-conopressin G | 11CON14 | Vasopressin-like peptide |
GsMTx4 | 08GSM001 | Mechano sensitive ion channels |
Obtustatin | 10OBT001 | Blocks the binding of α1β1 integrin to collagen IV |
Rho-Conotoxin TIA | CON022 | Blocks α1-adrenergic receptor |
公司简介
Smartox Biotechnology是全球唯一一家专门生产动物毒液多肽毒素，用于细胞离子通道功能研究的生物医药公司。多肽毒素在生物制药领域具有重要的使用价值。
Smartox Biotechnology于2009年由来自Grenoble神经科学研究所(Grenoble Institute of Neuroscience)的Michel De Waard博士创立。Smartox Biotechnology专门研究动物毒液，制作合成多种毒液中的多肽成分（常称为毒素）。De Waard博士研究离子通道与毒素多肽的关系，尤其是鉴定、开发毒素多肽作为治疗性分子或细胞穿透肽(cell penetrating peptides, CPP)。其研究团队在毒液分离，药理性活性肽鉴定、富半胱氨酸肽定性、制作和优化等方面具有独特、丰富的经验。2010年，Smartox Biotechnolgy被法国研究部(Ministry of Research)授予“新兴企业OSEO奖(OSEO prize for emerging businesses)”。
总之，Smartox Biotechnolgy提供一系列高质量、具开创价值的多肽毒素。这些化合物在离子通道 研究中具有高的亲和性和选择性，是相应领域科学研究理想的生物毒素提供商和贴心的合作伙伴。
本网站将在规定时间内给予删除等相关处理。
