MINT MINT, Dpt of Biology, University of Rome Tor Vergata MINT http://mint.bio.uniroma2.it/mint url>http://mint.bio.uniroma2.it/mint/ https://mint.bio.uniroma2.it MINT-[0-9]+ https://mint.bio.uniroma2.it/index.php/results-interactions/?id=${ac} The Molecular INTeraction database (MINT) is a relational database designed to store interactions between biological molecules. http://mint.bio.uniroma2.it http://mint.bio.uniroma2.it/mint anonymous-2014x-1 Identification of the synaptic vesicle glycoprotein 2 receptor binding site in botulinum neurotoxin A in vitro In vitro pull down pull down weight by comassie molecular weight estimation by coomasie staining weight by comassie Rummel.Andreas@mh-hannover.de 2014 imex curation Only protein-protein interactions anonymous-2014x-2 Identification of the synaptic vesicle glycoprotein 2 receptor binding site in botulinum neurotoxin A in vitro In vitro anti tag coip anti tag coimmunoprecipitation anti tag coip western blot western blot Immuno blot Immuno blot Rummel.Andreas@mh-hannover.de 2014 imex curation Only protein-protein interactions sv2c_rat Synaptic vesicle glycoprotein 2C Sv2c protein protein rat Rattus norvegicus (Rat) Rat MEDSYKDRTSLMKGAKDIAKEVKKQTVKKVNQAVDRAQDEYTQRSYSRFQDEDDDDDYYPPGETYSGEANDDEGSSEATEGHDEEDEIYEGEYQGIPSTNQGKDSIVSVGQPKGDEYKDRRELESERRADEEELAQQYELIIQECGHGRFQWALFFVLGMALMADGVEVFVVGFVLPSAETDLCIPNSGSGWLGSIVYLGMMVGAFFWGGLADKVGRKQSLLICMSVNGFFAFLSSFVQGYGFFLLCRLLSGFGIGGAIPTVFSYFAEVLAREKRGEHLSWLCMFWMIGGIYASAMAWAIIPHYGWSFSMGSAYQFHSWRVFVIVCALPCVSSVVALTFMPESPRFLLEVGKHDEAWMILKLIHDTNMRARGQPEKVFTVNKIKTPKQIDELIEIESDTGTWYRRCFVRIRTELYGIWLTFMRCFNYPVRENTIKLTIVWFTLSFGYYGLSVWFPDVIKHLQSDEYALLTRNVQKDKYANFSINFTMENQVHTGMEYDNGRFLGVKFKSVTFKDSVFKSCTFDDVTSVNTYFKNCTFIDTLFENTDFEPYKFIDSEFQNCSFLHNKTGCQITFDDDYSAYWIYFVNFLGTLAVLPGNIVSALLMDRIGRLTMLGGSMVLSGISCFFLWFGTSESMMIGMLCLYNGLTISAWNSLDVVTVELYPTDRRATGFGFLNALCKAAAVLGNLIFGSLVSITKAIPILLASTVLVCGGLVGLRLPDTRTQVLM mint Plays a role in the control of regulated secretion in neural and endocrine cells, enhancing selectively low-frequency neurotransmission. Positively regulates vesicle fusion by maintaining the readily releasable pool of secretory vesicles Receptor for the botulinium neurotoxin type A/BOTA bxa1_clobo Botulinum neurotoxin type A botA atx Bontoxilysin-A Botulinum neurotoxin type A1 bonT protein protein clobo Clostridium botulinum MPFVNKQFNYKDPVNGVDIAYIKIPNVGQMQPVKAFKIHNKIWVIPERDTFTNPEEGDLNPPPEAKQVPVSYYDSTYLSTDNEKDNYLKGVTKLFERIYSTDLGRMLLTSIVRGIPFWGGSTIDTELKVIDTNCINVIQPDGSYRSEELNLVIIGPSADIIQFECKSFGHEVLNLTRNGYGSTQYIRFSPDFTFGFEESLEVDTNPLLGAGKFATDPAVTLAHELIHAGHRLYGIAINPNRVFKVNTNAYYEMSGLEVSFEELRTFGGHDAKFIDSLQENEFRLYYYNKFKDIASTLNKAKSIVGTTASLQYMKNVFKEKYLLSEDTSGKFSVDKLKFDKLYKMLTEIYTEDNFVKFFKVLNRKTYLNFDKAVFKINIVPKVNYTIYDGFNLRNTNLAANFNGQNTEINNMNFTKLKNFTGLFEFYKLLCVRGIITSKTKSLDKGYNKALNDLCIKVNNWDLFFSPSEDNFTNDLNKGEEITSDTNIEAAEENISLDLIQQYYLTFNFDNEPENISIENLSSDIIGQLELMPNIERFPNGKKYELDKYTMFHYLRAQEFEHGKSRIALTNSVNEALLNPSRVYTFFSSDYVKKVNKATEAAMFLGWVEQLVYDFTDETSEVSTTDKIADITIIIPYIGPALNIGNMLYKDDFVGALIFSGAVILLEFIPEIAIPVLGTFALVSYIANKVLTVQTIDNALSKRNEKWDEVYKYIVTNWLAKVNTQIDLIRKKMKEALENQAEATKAIINYQYNQYTEEEKNNINFNIDDLSSKLNESINKAMININKFLNQCSVSYLMNSMIPYGVKRLEDFDASLKDALLKYIYDNRGTLIGQVDRLKDKVNNTLSTDIPFQLSKYVDNQRLLSTFTEYIKNIINTSILNLRYESNHLIDLSRYASKINIGSKVNFDPIDKNQIQLFNLESSKIEVILKNAIVYNSMYENFSTSFWIRIPKYFNSISLNNEYTIINCMENNSGWKVSLNYGEIIWTLQDTQEIKQRVVFKYSQMINISDYINRWIFVTITNNRLNNSKIYINGRLIDQKPISNLGNIHASNNIMFKLDGCRDTHRYIWIKYFNLFDKELNEKEIKDLYDNQSNSGILKDFWGDYLQYDKPYYMLNLYDPNKYVDVNNVGIRGYMYLKGPRGSVMTTNIYLNSSLYRGTKFIIKKYASGNKDNIVRNNDRVYINVVVKNKEYRLATNASQAGVEKILSALEIPDVGNLSQVVVMKSKNDQGITNKCKMNLQDNNGNDIGFIGFHQFNNIAKLVASNWYNRQIERSSRTLGCSWEFIPVDDGWGERPL mint Inhibits acetylcholine release. The botulinum toxin binds with high affinity to peripheral neuronal presynaptic membrane to the secretory vesicle protein SV2. It binds directly to the largest luminal loop of SV2A, SV2B and SV2C. It is then internalized by receptor-mediated endocytosis. The C-terminus of the heavy chain (H) is responsible for the adherence of the toxin to the cell surface while the N-terminus mediates transport of the light chain from the endocytic vesicle to the cytosol. After translocation, the light chain (L) hydrolyzes the 197-Gln-|-Arg-198 bond in SNAP-25, thereby blocking neurotransmitter release. Inhibition of acetylcholine release results in flaccid paralysis, with frequent heart or respiratory failure sv2a_rat Synaptic vesicle glycoprotein 2A Sv2a Sv2 protein protein rat Rattus norvegicus (Rat) Rat MEEGFRDRAAFIRGAKDIAKEVKKHAAKKVVKGLDRVQDEYSRRSYSRFEEEEDDDDFPAPADGYYRGEGAQDEEEGGASSDATEGHDEDDEIYEGEYQGIPRAESGGKGERMADGAPLAGVRGGLSDGEGPPGGRGEAQRRKDREELAQQYETILRECGHGRFQWTLYFVLGLALMADGVEVFVVGFVLPSAEKDMCLSDSNKGMLGLIVYLGMMVGAFLWGGLADRLGRRQCLLISLSVNSVFAFFSSFVQGYGTFLFCRLLSGVGIGGSIPIVFSYFSEFLAQEKRGEHLSWLCMFWMIGGVYAAAMAWAIIPHYGWSFQMGSAYQFHSWRVFVLVCAFPSVFAIGALTTQPESPRFFLENGKHDEAWMVLKQVHDTNMRAKGHPERVFSVTHIKTIHQEDELIEIQSDTGTWYQRWGVRALSLGGQVWGNFLSCFSPEYRRITLMMMGVWFTMSFSYYGLTVWFPDMIRHLQAVDYAARTKVFPGERVEHVTFNFTLENQIHRGGQYFNDKFIGLRLKSVSFEDSLFEECYFEDVTSSNTFFRNCTFINTVFYNTDLFEYKFVNSRLVNSTFLHNKEGCPLDVTGTGEGAYMVYFVSFLGTLAVLPGNIVSALLMDKIGRLRMLAGSSVLSCVSCFFLSFGNSESAMIALLCLFGGVSIASWNALDVLTVELYPSDKRTTAFGFLNALCKLAAVLGISIFTSFVGITKAAPILFASAALALGSSLALKLPETRGQVLQ sv2b_rat Synaptic vesicle glycoprotein 2B Sv2b Sv2bb protein protein rat Rattus norvegicus (Rat) Rat MDDYRYRDNYEGYAPNDGYYRGNEQNPEEDAQSDVTEGHDEEDEIYEGEYQGIPHPDDVKSKQTKMAPSRADGLRGQADLMAERMEDEEQLAHQYETIIDECGHGRFQWTLFFVLVLALMADGVEVFVVSFALPSAEKDMCLSSSKKGMLGLIVYLGMMAGAFILGGLADKLGRKKVLSMSLAINASFASLSSFVQGYGAFLFCRLISGIGIGGSLPIVFAYFSEFLSREKRGEHLSWLGIFWMTGGIYASAMAWSIIPHYGWGFSMGTNYHFHSWRVFVIVCALPATVSMVALKFMPESPRFLLEMGKHDEAWMILKQVHDTNMRAKGTPEKVFTVSHIKTPKQMDEFIEIQSSTGTWYQRWLVRFMTIFKQVWDNALYCVMGPYRMNTLILAVVWFTMALSYYGLTVWFPDMIRYFQDEEYKSKMKVFFGEHVHGATINFTMENQIHQHGKLVNDKFIKMYFKHVLFEDTFFDKCYFEDVTSTDTYFKNCTIESTTFYNTDLYKHKFIDCRFINSTFLEQKEGCHMDFEEDNDFLIYLVSFLGSLSVLPGNIISALLMDRIGRLKMIGGSMLISAVCCFFLFFGNSESAMIGWQCLFCGTSIAAWNALDVITVELYPTNQRATAFGILNGLCKLGAILGNTIFASFVGITKVVPILLAAASLVGGGLVALRLPETREQVLM bxe_clobo Botulinum neurotoxin type E botE Bontoxilysin-E protein protein clobo Clostridium botulinum MPKINSFNYNDPVNDRTILYIKPGGCQEFYKSFNIMKNIWIIPERNVIGTTPQDFHPPTSLKNGDSSYYDPNYLQSDEEKDRFLKIVTKIFNRINNNLSGGILLEELSKANPYLGNDNTPDNQFHIGDASAVEIKFSNGSQDILLPNVIIMGAEPDLFETNSSNISLRNNYMPSNHRFGSIAIVTFSPEYSFRFNDNCMNEFIQDPALTLMHELIHSLHGLYGAKGITTKYTITQKQNPLITNIRGTNIEEFLTFGGTDLNIITSAQSNDIYTNLLADYKKIASKLSKVQVSNPLLNPYKDVFEAKYGLDKDASGIYSVNINKFNDIFKKLYSFTEFDLRTKFQVKCRQTYIGQYKYFKLSNLLNDSIYNISEGYNINNLKVNFRGQNANLNPRIITPITGRGLVKKIIRFCKNIVSVKGIRKSICIEINNGELFFVASENSYNDDNINTPKEIDDTVTSNNNYENDLDQVILNFNSESAPGLSDEKLNLTIQNDAYIPKYDSNGTSDIEQHDVNELNVFFYLDAQKVPEGENNVNLTSSIDTALLEQPKIYTFFSSEFINNVNKPVQAALFVSWIQQVLVDFTTEANQKSTVDKIADISIVVPYIGLALNIGNEAQKGNFKDALELLGAGILLEFEPELLIPTILVFTIKSFLGSSDNKNKVIKAINNALKERDEKWKEVYSFIVSNWMTKINTQFNKRKEQMYQALQNQVNAIKTIIESKYNSYTLEEKNELTNKYDIKQIENELNQKVSIAMNNIDRFLTESSISYLMKIINEVKINKLREYDENVKTYLLNYIIQHGSILGESQQELNSMVTDTLNNSIPFKLSSYTDDKILISYFNKFFKRIKSSSVLNMRYKNDKYVDTSGYDSNININGDVYKYPTNKNQFGIYNDKLSEVNISQNDYIIYDNKYKNFSISFWVRIPNYDNKIVNVNNEYTIINCMRDNNSGWKVSLNHNEIIWTFEDNRGINQKLAFNYGNANGISDYINKWIFVTITNDRLGDSKLYINGNLIDQKSILNLGNIHVSDNILFKIVNCSYTRYIGIRYFNIFDKELDETEIQTLYSNEPNTNILKDFWGNYLLYDKEYYLLNVLKPNNFIDRRKDSTLSINNIRSTILLANRLYSGIKVKIQRVNNSSTNDNLVRKNDQVYINFVASKTHLFPLYADTATTNKEKTIKISSSGNRFNQVVVMNSVGNCTMNFKNNNGNNIGLLGFKADTVVASTWYYTHMRDHTNSNGCFWNFISEEHGWQEK bxf_clobo Botulinum neurotoxin type F botF Bontoxilysin-F protein protein clobo Clostridium botulinum MPVAINSFNYNDPVNDDTILYMQIPYEEKSKKYYKAFEIMRNVWIIPERNTIGTNPSDFDPPASLKNGSSAYYDPNYLTTDAEKDRYLKTTIKLFKRINSNPAGKVLLQEISYAKPYLGNDHTPIDEFSPVTRTTSVNIKLSTNVESSMLLNLLVLGAGPDIFESCCYPVRKLIDPDVVYDPSNYGFGSINIVTFSPEYEYTFNDISGGHNSSTESFIADPAISLAHELIHALHGLYGARGVTYEETIEVKQAPLMIAEKPIRLEEFLTFGGQDLNIITSAMKEKIYNNLLANYEKIATRLSEVNSAPPEYDINEYKDYFQWKYGLDKNADGSYTVNENKFNEIYKKLYSFTESDLANKFKVKCRNTYFIKYEFLKVPNLLDDDIYTVSEGFNIGNLAVNNRGQSIKLNPKIIDSIPDKGLVEKIVKFCKSVIPRKGTKAPPRLCIRVNNSELFFVASESSYNENDINTPKEIDDTTNLNNNYRNNLDEVILDYNSQTIPQISNRTLNTLVQDNSYVPRYDSNGTSEIEEYDVVDFNVFFYLHAQKVPEGETNISLTSSIDTALLEESKDIFFSSEFIDTINKPVNAALFIDWISKVIRDFTTEATQKSTVDKIADISLIVPYVGLALNIIIEAEKGNFEEAFELLGVGILLEFVPELTIPVILVFTIKSYIDSYENKNKAIKAINNSLIEREAKWKEIYSWIVSNWLTRINTQFNKRKEQMYQALQNQVDAIKTAIEYKYNNYTSDEKNRLESEYNINNIEEELNKKVSLAMKNIERFMTESSISYLMKLINEAKVGKLKKYDNHVKSDLLNYILDHRSILGEQTNELSDLVTSTLNSSIPFELSSYTNDKILIIYFNRLYKKIKDSSILDMRYENNKFIDISGYGSNISINGNVYIYSTNRNQFGIYNSRLSEVNIAQNNDIIYNSRYQNFSISFWVRIPKHYKPMNHNREYTIINCMGNNNSGWKISLRTVRDCEIIWTLQDTSGNKENLIFRYEELNRISNYINKWIFVTITNNRLGNSRIYINGNLIVEKSISNLGDIHVSDNILFKIVGCDDETYVGIRYFKVFNTELDKTEIETLYSNEPDPSILKNYWGNYLLYNKKYYLFNLLRKDKYITLNSGILNINQQRGVTEGSVFLNYKLYEGVEVIIRKNGPIDISNTDNFVRKNDLAYINVVDRGVEYRLYADTKSEKEKIIRTSNLNDSLGQIIVMDSIGNNCTMNFQNNNGSNIGLLGFHSNNLVASSWYYNNIRRNTSSNGCFWSSISKENGWKE bota-sv2c 1481148 n/a rSV2C 1481150 weight by comassie molecular weight estimation by coomasie staining weight by comassie unspecified role unspecified role bait bait purified purified gst tag gst tag glutathione s tranferase tag glutathione S-tranferase tag glutathione S-tranferase tag gst tag undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined false binding region binding region binding-associated region binding region certain certain sequence position certain certain certain sequence position certain false n/a BoNT/A 1481151 weight by comassie molecular weight estimation by coomasie staining weight by comassie unspecified role unspecified role prey prey purified purified his tag his tag his tag Hexa-His-tag 6-His-tag Histidine-tag Hexa-His-tag Histidine-tag 6-His-tag undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined false direct interaction direct interaction false false false f1a This direct protein-protein interaction is clearly demonstrated in a GST pull down assay employing full-length recombinant H6tBoNTA and rat GST-rSV2C 454-579 (Fig. 1A). The weaker binding of HCA to recombinant rat GST-rSV2A 468-594 is also shown (Fig. S1) whereas binding to non-glycosylated rSV2B is hardly detectable. Well characterised neutralising monoclonal antibodies directed against different epitopes within the HC-fragment of BoNT/A [25-29] were screened for interference with the SV2C interaction sv2c-bota-1 1481148 n/a HcA 1481151 weight by comassie molecular weight estimation by coomasie staining weight by comassie unspecified role unspecified role prey prey purified purified binding region binding region binding-associated region binding region certain certain sequence position certain certain certain sequence position certain false tag tandem tag tandem tag undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined false n/a rSV2A 1481152 weight by comassie molecular weight estimation by coomasie staining weight by comassie unspecified role unspecified role bait bait purified purified binding region binding region binding-associated region binding region certain certain sequence position certain certain certain sequence position certain false gst tag gst tag glutathione s tranferase tag glutathione S-tranferase tag glutathione S-tranferase tag gst tag undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined false direct interaction direct interaction false false false sf1 This direct protein-protein interaction is clearly demonstrated in a GST pull down assay employing full-length recombinant H6tBoNTA and rat GST-rSV2C 454-579 (Fig. 1A). The weaker binding of HCA to recombinant rat GST-rSV2A 468-594 is also shown (Fig. S1) whereas binding to non-glycosylated rSV2B is hardly detectable. Well characterised neutralising monoclonal antibodies directed against different epitopes within the HC-fragment of BoNT/A [25-29] were screened for interference with the SV2C interaction sv2c-bota-2 1481148 n/a rSV2C 1481150 predetermined predetermined participant predetermined unspecified role unspecified role bait bait purified purified binding region binding region binding-associated region binding region certain certain sequence position certain certain certain sequence position certain false gst tag gst tag glutathione s tranferase tag glutathione S-tranferase tag glutathione S-tranferase tag gst tag undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined false n/a BoNT/A 1481151 predetermined predetermined participant predetermined unspecified role unspecified role prey prey purified purified P0DPI0:p.Arg1131Glu mutation decreasing mutation decreasing interaction hotspot hotspot mutation decreasing certain certain sequence position certain certain certain sequence position certain false his tag his tag his tag Hexa-His-tag 6-His-tag Histidine-tag Hexa-His-tag Histidine-tag 6-His-tag undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined false direct interaction direct interaction false false false f2 This direct protein-protein interaction is clearly demonstrated in a GST pull down assay employing full-length recombinant H6tBoNTA and rat GST-rSV2C 454-579 (Fig. 1A). The weaker binding of HCA to recombinant rat GST-rSV2A 468-594 is also shown (Fig. S1) whereas binding to non-glycosylated rSV2B is hardly detectable. Well characterised neutralising monoclonal antibodies directed against different epitopes within the HC-fragment of BoNT/A [25-29] were screened for interference with the SV2C interaction sv2c-bota-3 1481148 n/a BoNT/A 1481151 predetermined predetermined participant predetermined unspecified role unspecified role prey prey purified purified his tag his tag his tag Hexa-His-tag 6-His-tag Histidine-tag Hexa-His-tag Histidine-tag 6-His-tag undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined false P0DPI0:p.Lys1163Glu mutation decreasing mutation decreasing interaction hotspot hotspot mutation decreasing certain certain sequence position certain certain certain sequence position certain false n/a rSV2C 1481150 predetermined predetermined participant predetermined unspecified role unspecified role bait bait purified purified gst tag gst tag glutathione s tranferase tag glutathione S-tranferase tag glutathione S-tranferase tag gst tag undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined false binding region binding region binding-associated region binding region certain certain sequence position certain certain certain sequence position certain false direct interaction direct interaction false false false f2 This direct protein-protein interaction is clearly demonstrated in a GST pull down assay employing full-length recombinant H6tBoNTA and rat GST-rSV2C 454-579 (Fig. 1A). The weaker binding of HCA to recombinant rat GST-rSV2A 468-594 is also shown (Fig. S1) whereas binding to non-glycosylated rSV2B is hardly detectable. Well characterised neutralising monoclonal antibodies directed against different epitopes within the HC-fragment of BoNT/A [25-29] were screened for interference with the SV2C interaction sv2c-bota-4 1481148 n/a BoNT/A 1481151 predetermined predetermined participant predetermined unspecified role unspecified role prey prey purified purified his tag his tag his tag Hexa-His-tag 6-His-tag Histidine-tag Hexa-His-tag Histidine-tag 6-His-tag undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined false P0DPI0:p.Asn1188Ala mutation decreasing mutation decreasing interaction hotspot hotspot mutation decreasing certain certain sequence position certain certain certain sequence position certain false n/a rSV2C 1481150 predetermined predetermined participant predetermined unspecified role unspecified role bait bait purified purified gst tag gst tag glutathione s tranferase tag glutathione S-tranferase tag glutathione S-tranferase tag gst tag undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined false binding region binding region binding-associated region binding region certain certain sequence position certain certain certain sequence position certain false direct interaction direct interaction false false false f2 This direct protein-protein interaction is clearly demonstrated in a GST pull down assay employing full-length recombinant H6tBoNTA and rat GST-rSV2C 454-579 (Fig. 1A). The weaker binding of HCA to recombinant rat GST-rSV2A 468-594 is also shown (Fig. S1) whereas binding to non-glycosylated rSV2B is hardly detectable. Well characterised neutralising monoclonal antibodies directed against different epitopes within the HC-fragment of BoNT/A [25-29] were screened for interference with the SV2C interaction sv2c-bota-5 1481148 n/a BoNT/A 1481151 predetermined predetermined participant predetermined unspecified role unspecified role prey prey purified purified P0DPI0:p.Asn905Tyr mutation decreasing mutation decreasing interaction hotspot hotspot mutation decreasing certain certain sequence position certain certain certain sequence position certain false his tag his tag his tag Hexa-His-tag 6-His-tag Histidine-tag Hexa-His-tag Histidine-tag 6-His-tag undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined false n/a rSV2C 1481150 predetermined predetermined participant predetermined unspecified role unspecified role bait bait purified purified binding region binding region binding-associated region binding region certain certain sequence position certain certain certain sequence position certain false gst tag gst tag glutathione s tranferase tag glutathione S-tranferase tag glutathione S-tranferase tag gst tag undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined false direct interaction direct interaction false false false f2 This direct protein-protein interaction is clearly demonstrated in a GST pull down assay employing full-length recombinant H6tBoNTA and rat GST-rSV2C 454-579 (Fig. 1A). The weaker binding of HCA to recombinant rat GST-rSV2A 468-594 is also shown (Fig. S1) whereas binding to non-glycosylated rSV2B is hardly detectable. Well characterised neutralising monoclonal antibodies directed against different epitopes within the HC-fragment of BoNT/A [25-29] were screened for interference with the SV2C interaction sv2c-bota-6 1481148 n/a BoNT/A 1481151 predetermined predetermined participant predetermined unspecified role unspecified role prey prey purified purified his tag his tag his tag Hexa-His-tag 6-His-tag Histidine-tag Hexa-His-tag Histidine-tag 6-His-tag undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined false P0DPI0:p.Thr1063Pro mutation decreasing mutation decreasing interaction hotspot hotspot mutation decreasing certain certain sequence position certain certain certain sequence position certain false n/a rSV2C 1481150 predetermined predetermined participant predetermined unspecified role unspecified role bait bait purified purified binding region binding region binding-associated region binding region certain certain sequence position certain certain certain sequence position certain false gst tag gst tag glutathione s tranferase tag glutathione S-tranferase tag glutathione S-tranferase tag gst tag undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined false direct interaction direct interaction false false false f2 This direct protein-protein interaction is clearly demonstrated in a GST pull down assay employing full-length recombinant H6tBoNTA and rat GST-rSV2C 454-579 (Fig. 1A). The weaker binding of HCA to recombinant rat GST-rSV2A 468-594 is also shown (Fig. S1) whereas binding to non-glycosylated rSV2B is hardly detectable. Well characterised neutralising monoclonal antibodies directed against different epitopes within the HC-fragment of BoNT/A [25-29] were screened for interference with the SV2C interaction sv2c-bota-7 1481148 n/a BoNT/A 1481151 predetermined predetermined participant predetermined unspecified role unspecified role prey prey purified purified P0DPI0:p.His1064Gly mutation decreasing mutation decreasing interaction hotspot hotspot mutation decreasing certain certain sequence position certain certain certain sequence position certain false his tag his tag his tag Hexa-His-tag 6-His-tag Histidine-tag Hexa-His-tag Histidine-tag 6-His-tag undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined false n/a rSV2C 1481150 predetermined predetermined participant predetermined unspecified role unspecified role bait bait purified purified gst tag gst tag glutathione s tranferase tag glutathione S-tranferase tag glutathione S-tranferase tag gst tag undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined false binding region binding region binding-associated region binding region certain certain sequence position certain certain certain sequence position certain false direct interaction direct interaction false false false f2 This direct protein-protein interaction is clearly demonstrated in a GST pull down assay employing full-length recombinant H6tBoNTA and rat GST-rSV2C 454-579 (Fig. 1A). The weaker binding of HCA to recombinant rat GST-rSV2A 468-594 is also shown (Fig. S1) whereas binding to non-glycosylated rSV2B is hardly detectable. Well characterised neutralising monoclonal antibodies directed against different epitopes within the HC-fragment of BoNT/A [25-29] were screened for interference with the SV2C interaction sv2c-bota-8 1481148 n/a rSV2C 1481150 predetermined predetermined participant predetermined unspecified role unspecified role bait bait purified purified gst tag gst tag glutathione s tranferase tag glutathione S-tranferase tag glutathione S-tranferase tag gst tag undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined false binding region binding region binding-associated region binding region certain certain sequence position certain certain certain sequence position certain false n/a BoNT/A 1481151 predetermined predetermined participant predetermined unspecified role unspecified role prey prey purified purified his tag his tag his tag Hexa-His-tag 6-His-tag Histidine-tag Hexa-His-tag Histidine-tag 6-His-tag undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined false P0DPI0:p.Asp1108Arg mutation decreasing mutation decreasing interaction hotspot hotspot mutation decreasing certain certain sequence position certain certain certain sequence position certain false direct interaction direct interaction false false false f2 This direct protein-protein interaction is clearly demonstrated in a GST pull down assay employing full-length recombinant H6tBoNTA and rat GST-rSV2C 454-579 (Fig. 1A). The weaker binding of HCA to recombinant rat GST-rSV2A 468-594 is also shown (Fig. S1) whereas binding to non-glycosylated rSV2B is hardly detectable. Well characterised neutralising monoclonal antibodies directed against different epitopes within the HC-fragment of BoNT/A [25-29] were screened for interference with the SV2C interaction sv2c-bota-9 1481148 n/a rSV2C 1481150 predetermined predetermined participant predetermined unspecified role unspecified role bait bait purified purified gst tag gst tag glutathione s tranferase tag glutathione S-tranferase tag glutathione S-tranferase tag gst tag undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined false binding region binding region binding-associated region binding region certain certain sequence position certain certain certain sequence position certain false n/a BoNT/A 1481151 predetermined predetermined participant predetermined unspecified role unspecified role prey prey purified purified his tag his tag his tag Hexa-His-tag 6-His-tag Histidine-tag Hexa-His-tag Histidine-tag 6-His-tag undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined false P0DPI0:p.Arg1156Met mutation decreasing mutation decreasing interaction hotspot hotspot mutation decreasing certain certain sequence position certain certain certain sequence position certain false direct interaction direct interaction false false false f2 This direct protein-protein interaction is clearly demonstrated in a GST pull down assay employing full-length recombinant H6tBoNTA and rat GST-rSV2C 454-579 (Fig. 1A). The weaker binding of HCA to recombinant rat GST-rSV2A 468-594 is also shown (Fig. S1) whereas binding to non-glycosylated rSV2B is hardly detectable. Well characterised neutralising monoclonal antibodies directed against different epitopes within the HC-fragment of BoNT/A [25-29] were screened for interference with the SV2C interaction sv2c-bota-10 1481148 n/a BoNT/A 1481151 predetermined predetermined participant predetermined unspecified role unspecified role prey prey purified purified P0DPI0:p.Val1287Glu mutation decreasing mutation decreasing interaction hotspot hotspot mutation decreasing certain certain sequence position certain certain certain sequence position certain false his tag his tag his tag Hexa-His-tag 6-His-tag Histidine-tag Hexa-His-tag Histidine-tag 6-His-tag undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined false n/a rSV2C 1481150 predetermined predetermined participant predetermined unspecified role unspecified role bait bait purified purified binding region binding region binding-associated region binding region certain certain sequence position certain certain certain sequence position certain false gst tag gst tag glutathione s tranferase tag glutathione S-tranferase tag glutathione S-tranferase tag gst tag undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined false direct interaction direct interaction false false false f2 This direct protein-protein interaction is clearly demonstrated in a GST pull down assay employing full-length recombinant H6tBoNTA and rat GST-rSV2C 454-579 (Fig. 1A). The weaker binding of HCA to recombinant rat GST-rSV2A 468-594 is also shown (Fig. S1) whereas binding to non-glycosylated rSV2B is hardly detectable. Well characterised neutralising monoclonal antibodies directed against different epitopes within the HC-fragment of BoNT/A [25-29] were screened for interference with the SV2C interaction sv2c-bota-11 1481148 n/a BoNT/A 1481151 predetermined predetermined participant predetermined unspecified role unspecified role prey prey purified purified P0DPI0:p.Gly1292Arg mutation decreasing mutation decreasing interaction hotspot hotspot mutation decreasing certain certain sequence position certain certain certain sequence position certain false his tag his tag his tag Hexa-His-tag 6-His-tag Histidine-tag Hexa-His-tag Histidine-tag 6-His-tag undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined false n/a rSV2C 1481150 predetermined predetermined participant predetermined unspecified role unspecified role bait bait purified purified binding region binding region binding-associated region binding region certain certain sequence position certain certain certain sequence position certain false gst tag gst tag glutathione s tranferase tag glutathione S-tranferase tag glutathione S-tranferase tag gst tag undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined false direct interaction direct interaction false false false f This direct protein-protein interaction is clearly demonstrated in a GST pull down assay employing full-length recombinant H6tBoNTA and rat GST-rSV2C 454-579 (Fig. 1A). The weaker binding of HCA to recombinant rat GST-rSV2A 468-594 is also shown (Fig. S1) whereas binding to non-glycosylated rSV2B is hardly detectable. Well characterised neutralising monoclonal antibodies directed against different epitopes within the HC-fragment of BoNT/A [25-29] were screened for interference with the SV2C interaction bota-sv2a 1481149 n/a HcA 1481151 western blot western blot Immuno blot Immuno blot unspecified role unspecified role bait bait purified purified tag tandem tag tandem tag undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined false binding region binding region binding-associated region binding region certain certain sequence position certain certain certain sequence position certain false P0DPI0:p.Gly1292Arg mutation disrupting mutation disrupting interaction mutation disrupting certain certain sequence position certain certain certain sequence position certain false n/a SV2B 1481153 western blot western blot Immuno blot Immuno blot unspecified role unspecified role prey prey subcellular prep subcellular preparation subcellular prep physiological level physiological level endogenous level endogenous endogenous endogenous level n/a SV2A 1481152 western blot western blot Immuno blot Immuno blot unspecified role unspecified role prey prey subcellular prep subcellular preparation subcellular prep physiological level physiological level endogenous level endogenous endogenous endogenous level physical association physical association false false false f4 HCA mutants R1156M and V1287E showed similar affinity to SV2A and SV2B as wild-type HCA indicating subtle differences between the mode of interaction of SV2A/B and of SV2C with BoNT/A bota-sv2a-1 1481149 n/a SV2B 1481153 western blot western blot Immuno blot Immuno blot unspecified role unspecified role prey prey subcellular prep subcellular preparation subcellular prep physiological level physiological level endogenous level endogenous endogenous endogenous level n/a SV2A 1481152 western blot western blot Immuno blot Immuno blot unspecified role unspecified role prey prey subcellular prep subcellular preparation subcellular prep physiological level physiological level endogenous level endogenous endogenous endogenous level n/a HcE 1481154 western blot western blot Immuno blot Immuno blot unspecified role unspecified role bait bait purified purified binding region binding region binding-associated region binding region certain certain sequence position certain certain certain sequence position certain false tag tandem tag tandem tag undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined false physical association physical association false false false f4 HCA mutants R1156M and V1287E showed similar affinity to SV2A and SV2B as wild-type HCA indicating subtle differences between the mode of interaction of SV2A/B and of SV2C with BoNT/A bxe-sv2a 1481149 n/a HcF 1481155 western blot western blot Immuno blot Immuno blot unspecified role unspecified role bait bait purified purified binding region binding region binding-associated region binding region certain certain sequence position certain certain certain sequence position certain false tag tandem tag tandem tag undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined false n/a SV2A 1481152 western blot western blot Immuno blot Immuno blot unspecified role unspecified role prey prey subcellular prep subcellular preparation subcellular prep physiological level physiological level endogenous level endogenous endogenous endogenous level n/a SV2B 1481153 western blot western blot Immuno blot Immuno blot unspecified role unspecified role prey prey subcellular prep subcellular preparation subcellular prep physiological level physiological level endogenous level endogenous endogenous endogenous level physical association physical association false false false f4 HCA mutants R1156M and V1287E showed similar affinity to SV2A and SV2B as wild-type HCA indicating subtle differences between the mode of interaction of SV2A/B and of SV2C with BoNT/A