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Smartox/Selective blocker of TTX-S VGSC/13HTX003-00500/0.5mg

Hainantoxin-III(HNTX-III;hainantoxin-3) isapeptidethathasbeenisolatedfromthevenomoftheChinesebirdspider Seleconosmiahainana. Hainantoxin-III specifically blocksmammalianneuronaltetrodotoxin-sensitivevoltage-gatedsodiumchannels (VGSCs). HainantoxinIII wasfoundinactiveontetrodotoxin-resistantVGSCsandvoltage-gatedCa²⁺channels(bothhighandlowvoltage-activated).
Hainantoxin-III stronglydepressedtheamplitudeofratDRGtetrodotoxin-sensitiveNa⁺ currentswithanIC₅₀ valueof1.1nM.Like Hainantoxin-IV, Hainantoxin-III causesahyperpolarizingshiftofabout10mVinthevoltagemidpointofsteady-stateNa⁺ channelinactivation.Similarto Huwentoxin-IV, Hainantoxin-III and Hainantoxin-IV donotaffecttheactivationandinactivationkineticsofNa⁺ currents.
Hainantoxin-IIIinhibitsNa_v1.7 currentamplitudewithoutsignificantlyalteringtheactivationandinactivationkinetics.Hainantoxin-III increasesthedeactivationoftheNa_v1.7currentafterextremedepolarizations. Hainantoxin-III seemstointeractwithsite4andtotrapthedomainIIvoltagesensorintheclosedstate.TheinhibitionofNa_v1.7byhainantoxin-IIIisreversIBLeuponwashing,butnoreversibilitywasobservedfor Hainantoxin-IV and Huwentoxin-IV. Hainantoxin-IIIwasshowntoblock Na_v1.1,Na_v1.2,Na_v1.3andNa_v1.7 expressedinHEK293cellswithIC₅₀ valuesof1.27µM,275nM,491nMand232nM,respectively.

Description:

Productcode:N/A.Category:Sodiumchannels.Tags:nav,tetrodotoxin,ttx.

AAsequence: Gly-Cys²-Lys-Gly-Phe-Gly-Asp-Ser-Cys⁹-Thr-Pro-Gly-Lys-Asn-Glu-Cys¹⁶-Cys¹⁷-Pro-Asn-Tyr-Ala-Cys²²-Ser-Ser-Lys-His-Lys-Trp-Cys²⁹-Lys-Val-Tyr-Leu-NH₂
Disulfidebonds: Cys²-Cys¹⁷,Cys⁹-Cys²²,andCys¹⁶-Cys²⁹
Length(aa): 33
Formula: C₁₅₄H₂₂₈N₄₄O₄₅S₆
MolecularWeight: 3608.20Da
Appearance:Whitelyophilizedsolid
Solubility: waterandsalinebuffer
CASnumber: Notavailable
Source: Synthetic
Purityrate: >97%

Reference:

Inhibitionofneuronaltetrodotoxin-sensitiveNa+channelsbytwospidertoxins:hainantoxin-IIIandhainantoxin-IV.

Hainantoxin-IIIandhainantoxin-IV,isolatedfromthevenomoftheChinesebirdspiderSeleconosmiahainana,areneurotoxicpeptidescomposedof33-35residueswiththreedisulfidebonds.Usingwhole-cellpatch-clamptechnique,weinvestigatedtheiractiononionicchannelsofadultratdorsalrootganglionneurons.ItwasfoundthatthetwotoxinsdidnotaffectCa2+channels(bothhighvoltageactivatedandlowvoltageactivatedtypes)nortetrodotoxin-resistantvoltage-gatedNa+channels(VGSCs).However,hainantoxin-IIIandhainantoxin-IVstronglydepressedtheamplitudeoftetrodotoxin-sensitiveNa+currentswithIC50valuesof1.1and44.6nM,respectively.Bothhainantoxin-III(1nM)andhainantoxin-IV(50nM)causedahyperpolarizingshiftofabout10mVinthevoltagemidpointofsteady-stateNa+channelinactivation,buttheyshoweddifferenceinthereprimekineticsofVGSCs:hainantoxin-IIIsignificantlydecreasedtherecoveryratefrominactivationataprepulsepotentialof-80mVwhilehainantoxin-IVdidnotdo.Itisinterestingtonotethatsimilartohuwentoxin-IV,thetwohainantoxinsdidnotaffecttheactivationandinactivationkineticsofNa+currentsandataconcentrationof1microMtheycompletelyinhibitedtheslowinginactivationcurrentsinducedbyBMK-I(toxinIfromthescorpionButhusmartensiKarsch),ascorpionalpha-liketoxin.Theresultsindicatethathainantoxin-IIIandhainantoxin-IVarenovelspidertoxinsandaffectthemammalneuralNa+channelsthroughamechanismquitedifferentfromotherspidertoxinstargetingtheneuralreceptorsite3,suchasdelta-aractoxinsandmu-agatoxins.

XiaoY, etal.(2003)Inhibitionofneuronaltetrodotoxin-sensitiveNa+channelsbytwospidertoxins:hainantoxin-IIIandhainantoxin-IV. EurJPharmacol. PMID: 14512091

StructureandFunctionofHainantoxin-III,aSelectiveAntagoNISTofNeuronalTetrodotoxin-sensitiveVoltage-gatedSodiumChannelsIsolatedfromtheChineseBirdSpiderOrnithoctonushainana

Inthepresentstudy,weinvestigatedthestructureandfunctionofhainantoxin-III(HNTX-III),a33-residuepolypeptidefromthevenomofthespiderOrnithoctonushainana.Itisaselectiveantagonistofneuronaltetrodotoxin-sensitivevoltage-gatedsodiumchannels.HNTX-IIIsuppressedNav1.7currentamplitudewithoutsignificantlyalteringtheactivation,inactivation,andreprimingkinetics.Shortextremedepolarizationspartiallyactivatedthetoxin-boundchannel,indicatingvoltage-dependentinhibitionofHNTX-III.HNTX-IIIincreasedthedeactivationoftheNav1.7currentafterextremedepolarizations.TheHNTX-III·Nav1.7complexwasgraduallydissociateduponprolongedstrongdepolarizationsinavoltage-dependentmanner,andtheunboundtoxinreboundtoNav1.7afteralongrepolarization.Moreover,analysisofchimericchannelsshowedthattheDIIS3-S4linkerwascriticalforHNTX-IIIbindingtoNav1.7.ThesedataareconsistentwithHNTX-IIIinteractingwithNav1.7site4andtrappingthedomainIIvoltagesensorintheclosedstate.ThesolutionstructureofHNTX-IIIwasdeterminedbytwo-dimensionalNMRandshowntopossessaninhibitorcystineknotmotif.Structuralanalysisindicatedthatcertainbasic,hydrophobic,andaromaticresiduesmainlylocalizedintheCterminusmayconstituteanamphiphilicsurfacepotentiallyinvolvedinHNTX-IIIbindingtoNav1.7.Takentogether,ourresultsshowthatHNTX-IIIisdistinctfromβ-scorpiontoxinsandotherβ-spidertoxinsinitsmechanismofactionandbindingspecificityandaffinity.Thepresentfindingscontributetoourunderstandingofthemechanismoftoxin-sodiumchannelinteractionandprovideausefultoolfortheinvestigationofthestructureandfunctionofsodiumchannelisoformsandforthedevelopmentofanalgesics.

LiuZ., etal. (2013)StructureandFunctionofHainantoxin-III,aSelectiveAntagonistofNeuronalTetrodotoxin-sensitiveVoltage-gatedSodiumChannelsIsolatedfromtheChineseBirdSpiderOrnithoctonushainana. JBC. PMID: 23703613

Determinationofdisulfidebridgesoftwospidertoxins:Hainantoxin-IIIandHainantoxin-IV

Peptidetoxinsareusuallyhighlybridgedproteinswithmultipairsofintrachaindisulfidebonds.Analysisofdisulfideconnectivityisanimportantfacetofproteinstructuredetermination.Inthispaper,wesuccessfullyassignedthedisulfidelinkageoftwonovelpeptidetoxins,calledHNTX-IIIandHNTX-IV,isolatedfromthevenomofOrnithoctonushainanaspider.BothpeptidesareusefulinhibitorsofTTX-sensitivevoltage-gatedsodiumchannelsandarecomposedofsixcysteineresiduesthatformthreedisulfidebonds,respectively.Firstly,thepeptideswerepartiallyreducedbytris(2-carboxyethyl)-phosphine(TCEP)in0.1Mcitratebuffercontaining6Mguanidine-HClat40°Cfortenminutes.Subsequently,thepartiallyreducedintermediatescontainingfreethiolswereseparatedbyreversed-phasehigh-performanceliquidchromatography(RP-HPLC)andalkylatedbyrapidcarboxamidomethylation.Then,thedisulfidebondsoftheintermediateswereanalyzedbyEdmandegradation.Byusingthestrategyabove,disulfidelinkagesofHNTX-IIIandHNTX-IVweredeterminedasI-IV,II-VandIII-VIpattern.Inaddition,thisstudyalsoshowedthatthismethodmayhaveagreatpotentialfordeterminingthedisulfidebondsofspiderpeptidetoxins.