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 Identification of the synaptic vesicle glycoprotein 2 receptor binding site in botulinum neurotoxin A Identification of the synaptic vesicle glycoprotein 2 receptor binding site in botulinum neurotoxin A 2014 imex curation Rummel.Andreas@mh-hannover.de 10-Mar-2014: Contacted by IntAct-Help. Only protein-protein interactions 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 Accepted 2014-FEB-06 by LUANA 2014 imex curation Identification of the synaptic vesicle glycoprotein 2 receptor binding site in botulinum neurotoxin A Identification of the synaptic vesicle glycoprotein 2 receptor binding site in botulinum neurotoxin A 2014 imex curation Rummel.Andreas@mh-hannover.de 10-Mar-2014: Contacted by IntAct-Help. Only protein-protein interactions 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 Accepted 2014-FEB-06 by LUANA 2014 imex curation bxa1_clobo Botulinum neurotoxin type A botA atx Bontoxilysin-A Botulinum neurotoxin type A1 bonT protein protein clobo Clostridium botulinum MPFVNKQFNYKDPVNGVDIAYIKIPNVGQMQPVKAFKIHNKIWVIPERDTFTNPEEGDLNPPPEAKQVPVSYYDSTYLSTDNEKDNYLKGVTKLFERIYSTDLGRMLLTSIVRGIPFWGGSTIDTELKVIDTNCINVIQPDGSYRSEELNLVIIGPSADIIQFECKSFGHEVLNLTRNGYGSTQYIRFSPDFTFGFEESLEVDTNPLLGAGKFATDPAVTLAHELIHAGHRLYGIAINPNRVFKVNTNAYYEMSGLEVSFEELRTFGGHDAKFIDSLQENEFRLYYYNKFKDIASTLNKAKSIVGTTASLQYMKNVFKEKYLLSEDTSGKFSVDKLKFDKLYKMLTEIYTEDNFVKFFKVLNRKTYLNFDKAVFKINIVPKVNYTIYDGFNLRNTNLAANFNGQNTEINNMNFTKLKNFTGLFEFYKLLCVRGIITSKTKSLDKGYNKALNDLCIKVNNWDLFFSPSEDNFTNDLNKGEEITSDTNIEAAEENISLDLIQQYYLTFNFDNEPENISIENLSSDIIGQLELMPNIERFPNGKKYELDKYTMFHYLRAQEFEHGKSRIALTNSVNEALLNPSRVYTFFSSDYVKKVNKATEAAMFLGWVEQLVYDFTDETSEVSTTDKIADITIIIPYIGPALNIGNMLYKDDFVGALIFSGAVILLEFIPEIAIPVLGTFALVSYIANKVLTVQTIDNALSKRNEKWDEVYKYIVTNWLAKVNTQIDLIRKKMKEALENQAEATKAIINYQYNQYTEEEKNNINFNIDDLSSKLNESINKAMININKFLNQCSVSYLMNSMIPYGVKRLEDFDASLKDALLKYIYDNRGTLIGQVDRLKDKVNNTLSTDIPFQLSKYVDNQRLLSTFTEYIKNIINTSILNLRYESNHLIDLSRYASKINIGSKVNFDPIDKNQIQLFNLESSKIEVILKNAIVYNSMYENFSTSFWIRIPKYFNSISLNNEYTIINCMENNSGWKVSLNYGEIIWTLQDTQEIKQRVVFKYSQMINISDYINRWIFVTITNNRLNNSKIYINGRLIDQKPISNLGNIHASNNIMFKLDGCRDTHRYIWIKYFNLFDKELNEKEIKDLYDNQSNSGILKDFWGDYLQYDKPYYMLNLYDPNKYVDVNNVGIRGYMYLKGPRGSVMTTNIYLNSSLYRGTKFIIKKYASGNKDNIVRNNDRVYINVVVKNKEYRLATNASQAGVEKILSALEIPDVGNLSQVVVMKSKNDQGITNKCKMNLQDNNGNDIGFIGFHQFNNIAKLVASNWYNRQIERSSRTLGCSWEFIPVDDGWGERPL DIP protein P10845 original sequence version: 156 DIP protein P10845 original sequence version: 139 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 B731EF5BA5E62FDA sv2a_rat Synaptic vesicle glycoprotein 2A Sv2a Sv2 protein protein rat Rattus norvegicus (Rat) Rat MEEGFRDRAAFIRGAKDIAKEVKKHAAKKVVKGLDRVQDEYSRRSYSRFEEEEDDDDFPAPADGYYRGEGAQDEEEGGASSDATEGHDEDDEIYEGEYQGIPRAESGGKGERMADGAPLAGVRGGLSDGEGPPGGRGEAQRRKDREELAQQYETILRECGHGRFQWTLYFVLGLALMADGVEVFVVGFVLPSAEKDMCLSDSNKGMLGLIVYLGMMVGAFLWGGLADRLGRRQCLLISLSVNSVFAFFSSFVQGYGTFLFCRLLSGVGIGGSIPIVFSYFSEFLAQEKRGEHLSWLCMFWMIGGVYAAAMAWAIIPHYGWSFQMGSAYQFHSWRVFVLVCAFPSVFAIGALTTQPESPRFFLENGKHDEAWMVLKQVHDTNMRAKGHPERVFSVTHIKTIHQEDELIEIQSDTGTWYQRWGVRALSLGGQVWGNFLSCFSPEYRRITLMMMGVWFTMSFSYYGLTVWFPDMIRHLQAVDYAARTKVFPGERVEHVTFNFTLENQIHRGGQYFNDKFIGLRLKSVSFEDSLFEECYFEDVTSSNTFFRNCTFINTVFYNTDLFEYKFVNSRLVNSTFLHNKEGCPLDVTGTGEGAYMVYFVSFLGTLAVLPGNIVSALLMDKIGRLRMLAGSSVLSCVSCFFLSFGNSESAMIALLCLFGGVSIASWNALDVLTVELYPSDKRTTAFGFLNALCKLAAVLGISIFTSFVGITKAAPILFASAALALGSSLALKLPETRGQVLQ E10FC62BEEFE316A sv2b_rat Synaptic vesicle glycoprotein 2B Sv2b Sv2bb protein protein rat Rattus norvegicus (Rat) Rat MDDYRYRDNYEGYAPNDGYYRGNEQNPEEDAQSDVTEGHDEEDEIYEGEYQGIPHPDDVKSKQTKMAPSRADGLRGQADLMAERMEDEEQLAHQYETIIDECGHGRFQWTLFFVLVLALMADGVEVFVVSFALPSAEKDMCLSSSKKGMLGLIVYLGMMAGAFILGGLADKLGRKKVLSMSLAINASFASLSSFVQGYGAFLFCRLISGIGIGGSLPIVFAYFSEFLSREKRGEHLSWLGIFWMTGGIYASAMAWSIIPHYGWGFSMGTNYHFHSWRVFVIVCALPATVSMVALKFMPESPRFLLEMGKHDEAWMILKQVHDTNMRAKGTPEKVFTVSHIKTPKQMDEFIEIQSSTGTWYQRWLVRFMTIFKQVWDNALYCVMGPYRMNTLILAVVWFTMALSYYGLTVWFPDMIRYFQDEEYKSKMKVFFGEHVHGATINFTMENQIHQHGKLVNDKFIKMYFKHVLFEDTFFDKCYFEDVTSTDTYFKNCTIESTTFYNTDLYKHKFIDCRFINSTFLEQKEGCHMDFEEDNDFLIYLVSFLGSLSVLPGNIISALLMDRIGRLKMIGGSMLISAVCCFFLFFGNSESAMIGWQCLFCGTSIAAWNALDVITVELYPTNQRATAFGILNGLCKLGAILGNTIFASFVGITKVVPILLAAASLVGGGLVALRLPETREQVLM BF862EFD4B99EA7B bxf_clobo Botulinum neurotoxin type F botF Bontoxilysin-F protein protein clobo Clostridium botulinum MPVAINSFNYNDPVNDDTILYMQIPYEEKSKKYYKAFEIMRNVWIIPERNTIGTNPSDFDPPASLKNGSSAYYDPNYLTTDAEKDRYLKTTIKLFKRINSNPAGKVLLQEISYAKPYLGNDHTPIDEFSPVTRTTSVNIKLSTNVESSMLLNLLVLGAGPDIFESCCYPVRKLIDPDVVYDPSNYGFGSINIVTFSPEYEYTFNDISGGHNSSTESFIADPAISLAHELIHALHGLYGARGVTYEETIEVKQAPLMIAEKPIRLEEFLTFGGQDLNIITSAMKEKIYNNLLANYEKIATRLSEVNSAPPEYDINEYKDYFQWKYGLDKNADGSYTVNENKFNEIYKKLYSFTESDLANKFKVKCRNTYFIKYEFLKVPNLLDDDIYTVSEGFNIGNLAVNNRGQSIKLNPKIIDSIPDKGLVEKIVKFCKSVIPRKGTKAPPRLCIRVNNSELFFVASESSYNENDINTPKEIDDTTNLNNNYRNNLDEVILDYNSQTIPQISNRTLNTLVQDNSYVPRYDSNGTSEIEEYDVVDFNVFFYLHAQKVPEGETNISLTSSIDTALLEESKDIFFSSEFIDTINKPVNAALFIDWISKVIRDFTTEATQKSTVDKIADISLIVPYVGLALNIIIEAEKGNFEEAFELLGVGILLEFVPELTIPVILVFTIKSYIDSYENKNKAIKAINNSLIEREAKWKEIYSWIVSNWLTRINTQFNKRKEQMYQALQNQVDAIKTAIEYKYNNYTSDEKNRLESEYNINNIEEELNKKVSLAMKNIERFMTESSISYLMKLINEAKVGKLKKYDNHVKSDLLNYILDHRSILGEQTNELSDLVTSTLNSSIPFELSSYTNDKILIIYFNRLYKKIKDSSILDMRYENNKFIDISGYGSNISINGNVYIYSTNRNQFGIYNSRLSEVNIAQNNDIIYNSRYQNFSISFWVRIPKHYKPMNHNREYTIINCMGNNNSGWKISLRTVRDCEIIWTLQDTSGNKENLIFRYEELNRISNYINKWIFVTITNNRLGNSRIYINGNLIVEKSISNLGDIHVSDNILFKIVGCDDETYVGIRYFKVFNTELDKTEIETLYSNEPDPSILKNYWGNYLLYNKKYYLFNLLRKDKYITLNSGILNINQQRGVTEGSVFLNYKLYEGVEVIIRKNGPIDISNTDNFVRKNDLAYINVVDRGVEYRLYADTKSEKEKIIRTSNLNDSLGQIIVMDSIGNNCTMNFQNNNGSNIGLLGFHSNNLVASSWYYNNIRRNTSSNGCFWSSISKENGWKE 5B99756A7438B921 bxe_clobo Botulinum neurotoxin type E botE Bontoxilysin-E protein protein clobo Clostridium botulinum MPKINSFNYNDPVNDRTILYIKPGGCQEFYKSFNIMKNIWIIPERNVIGTTPQDFHPPTSLKNGDSSYYDPNYLQSDEEKDRFLKIVTKIFNRINNNLSGGILLEELSKANPYLGNDNTPDNQFHIGDASAVEIKFSNGSQDILLPNVIIMGAEPDLFETNSSNISLRNNYMPSNHRFGSIAIVTFSPEYSFRFNDNCMNEFIQDPALTLMHELIHSLHGLYGAKGITTKYTITQKQNPLITNIRGTNIEEFLTFGGTDLNIITSAQSNDIYTNLLADYKKIASKLSKVQVSNPLLNPYKDVFEAKYGLDKDASGIYSVNINKFNDIFKKLYSFTEFDLRTKFQVKCRQTYIGQYKYFKLSNLLNDSIYNISEGYNINNLKVNFRGQNANLNPRIITPITGRGLVKKIIRFCKNIVSVKGIRKSICIEINNGELFFVASENSYNDDNINTPKEIDDTVTSNNNYENDLDQVILNFNSESAPGLSDEKLNLTIQNDAYIPKYDSNGTSDIEQHDVNELNVFFYLDAQKVPEGENNVNLTSSIDTALLEQPKIYTFFSSEFINNVNKPVQAALFVSWIQQVLVDFTTEANQKSTVDKIADISIVVPYIGLALNIGNEAQKGNFKDALELLGAGILLEFEPELLIPTILVFTIKSFLGSSDNKNKVIKAINNALKERDEKWKEVYSFIVSNWMTKINTQFNKRKEQMYQALQNQVNAIKTIIESKYNSYTLEEKNELTNKYDIKQIENELNQKVSIAMNNIDRFLTESSISYLMKIINEVKINKLREYDENVKTYLLNYIIQHGSILGESQQELNSMVTDTLNNSIPFKLSSYTDDKILISYFNKFFKRIKSSSVLNMRYKNDKYVDTSGYDSNININGDVYKYPTNKNQFGIYNDKLSEVNISQNDYIIYDNKYKNFSISFWVRIPNYDNKIVNVNNEYTIINCMRDNNSGWKVSLNHNEIIWTFEDNRGINQKLAFNYGNANGISDYINKWIFVTITNDRLGDSKLYINGNLIDQKSILNLGNIHVSDNILFKIVNCSYTRYIGIRYFNIFDKELDETEIQTLYSNEPNTNILKDFWGNYLLYDKEYYLLNVLKPNNFIDRRKDSTLSINNIRSTILLANRLYSGIKVKIQRVNNSSTNDNLVRKNDQVYINFVASKTHLFPLYADTATTNKEKTIKISSSGNRFNQVVVMNSVGNCTMNFKNNNGNNIGLLGFKADTVVASTWYYTHMRDHTNSNGCFWNFISEEHGWQEK DIP protein Q00496 original sequence version: 85 DIP protein Q00496 original sequence version: 112 B05AA65FF1B30362 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 4C154C69341D8DB7 bota-sv2c 2 rSV2C 8 weight by comassie molecular weight estimation by coomasie staining weight by comassie 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 binding region binding region binding-associated region binding region certain certain sequence position certain certain certain sequence position certain BoNT/A 3 weight by comassie molecular weight estimation by coomasie staining weight by comassie 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 direct interaction direct interaction 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 2 HcA 3 weight by comassie molecular weight estimation by coomasie staining weight by comassie 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 tag tandem tag tandem tag undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined rSV2A 4 weight by comassie molecular weight estimation by coomasie staining weight by comassie 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 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 direct interaction direct interaction 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 2 rSV2C 8 predetermined predetermined participant predetermined 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 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 BoNT/A 3 predetermined predetermined participant predetermined weight by comassie molecular weight estimation by coomasie staining weight by comassie 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 R E Wed May 02 17:43:49 BST 2018 This feature has been corrected as a result of our quality control procedures. The original label was 'arg1131glu' Tue Jun 30 16:12:33 BST 2020 This feature has been corrected as a result of our quality control procedures. The original label was 'p.Arg1131Glu' 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 direct interaction direct interaction 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 2 BoNT/A 3 predetermined predetermined participant predetermined 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 P0DPI0:p.Lys1163Glu mutation decreasing mutation decreasing interaction hotspot hotspot mutation decreasing certain certain sequence position certain certain certain sequence position certain K E Wed May 02 18:19:04 BST 2018 This feature has been corrected as a result of our quality control procedures. The original label was 'lys1163glu' Tue Jun 30 18:01:02 BST 2020 This feature has been corrected as a result of our quality control procedures. The original label was 'p.Lys1163Glu' rSV2C 8 predetermined predetermined participant predetermined 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 binding region binding region binding-associated region binding region certain certain sequence position certain certain certain sequence position certain direct interaction direct interaction 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 2 BoNT/A 3 predetermined predetermined participant predetermined 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 P0DPI0:p.Asn1188Ala mutation decreasing mutation decreasing interaction hotspot hotspot mutation decreasing certain certain sequence position certain certain certain sequence position certain N A Wed May 02 20:25:56 BST 2018 This feature has been corrected as a result of our quality control procedures. The original label was 'asn1188ala' Tue Jun 30 20:18:10 BST 2020 This feature has been corrected as a result of our quality control procedures. The original label was 'p.Asn1188Ala' rSV2C 8 predetermined predetermined participant predetermined 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 binding region binding region binding-associated region binding region certain certain sequence position certain certain certain sequence position certain direct interaction direct interaction 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 2 BoNT/A 3 predetermined predetermined participant predetermined weight by comassie molecular weight estimation by coomasie staining weight by comassie 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 N Y Wed May 02 18:35:51 BST 2018 This feature has been corrected as a result of our quality control procedures. The original label was 'asn905tyr' Tue Jun 30 19:35:43 BST 2020 This feature has been corrected as a result of our quality control procedures. The original label was 'p.Asn905Tyr' 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 rSV2C 8 predetermined predetermined participant predetermined 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 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 direct interaction direct interaction 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 2 BoNT/A 3 predetermined predetermined participant predetermined 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 P0DPI0:p.Thr1063Pro mutation decreasing mutation decreasing interaction hotspot hotspot mutation decreasing certain certain sequence position certain certain certain sequence position certain T P Wed May 02 16:38:38 BST 2018 This feature has been corrected as a result of our quality control procedures. The original label was 'thr1063pro' Tue Jun 30 18:01:06 BST 2020 This feature has been corrected as a result of our quality control procedures. The original label was 'p.Thr1063Pro' rSV2C 8 predetermined predetermined participant predetermined 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 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 direct interaction direct interaction 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 2 BoNT/A 3 predetermined predetermined participant predetermined weight by comassie molecular weight estimation by coomasie staining weight by comassie 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 H G Wed May 02 15:36:44 BST 2018 This feature has been corrected as a result of our quality control procedures. The original label was 'his1064gly' Tue Jun 30 15:48:25 BST 2020 This feature has been corrected as a result of our quality control procedures. The original label was 'p.His1064Gly' 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 rSV2C 8 predetermined predetermined participant predetermined 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 binding region binding region binding-associated region binding region certain certain sequence position certain certain certain sequence position certain direct interaction direct interaction 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 2 rSV2C 8 predetermined predetermined participant predetermined 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 binding region binding region binding-associated region binding region certain certain sequence position certain certain certain sequence position certain BoNT/A 3 predetermined predetermined participant predetermined 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 P0DPI0:p.Asp1108Arg mutation decreasing mutation decreasing interaction hotspot hotspot mutation decreasing certain certain sequence position certain certain certain sequence position certain D R Wed May 02 17:23:47 BST 2018 This feature has been corrected as a result of our quality control procedures. The original label was 'asp1108arg' Tue Jun 30 19:22:05 BST 2020 This feature has been corrected as a result of our quality control procedures. The original label was 'p.Asp1108Arg' direct interaction direct interaction 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 2 rSV2C 8 predetermined predetermined participant predetermined 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 binding region binding region binding-associated region binding region certain certain sequence position certain certain certain sequence position certain BoNT/A 3 predetermined predetermined participant predetermined 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 P0DPI0:p.Arg1156Met mutation decreasing mutation decreasing interaction hotspot hotspot mutation decreasing certain certain sequence position certain certain certain sequence position certain R M Wed May 02 17:36:46 BST 2018 This feature has been corrected as a result of our quality control procedures. The original label was 'arg1156met' Tue Jun 30 18:35:09 BST 2020 This feature has been corrected as a result of our quality control procedures. The original label was 'p.Arg1156Met' direct interaction direct interaction 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 2 BoNT/A 3 predetermined predetermined participant predetermined weight by comassie molecular weight estimation by coomasie staining weight by comassie 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 V E Wed May 02 16:18:08 BST 2018 This feature has been corrected as a result of our quality control procedures. The original label was 'val1287glu' Tue Jun 30 17:00:48 BST 2020 This feature has been corrected as a result of our quality control procedures. The original label was 'p.Val1287Glu' 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 rSV2C 8 predetermined predetermined participant predetermined 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 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 direct interaction direct interaction 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 2 BoNT/A 3 predetermined predetermined participant predetermined weight by comassie molecular weight estimation by coomasie staining weight by comassie 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 G R Wed May 02 19:16:18 BST 2018 This feature has been corrected as a result of our quality control procedures. The original label was 'gly1292arg' Tue Jun 30 17:53:21 BST 2020 This feature has been corrected as a result of our quality control procedures. The original label was 'p.Gly1292Arg' 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 rSV2C 8 predetermined predetermined participant predetermined 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 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 direct interaction direct interaction 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 1 HcA 3 western blot western blot Immuno blot Immuno blot 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 binding region binding region binding-associated region binding region certain certain sequence position certain certain certain sequence position certain P0DPI0:p.Gly1292Arg mutation disrupting mutation disrupting interaction mutation disrupting certain certain sequence position certain certain certain sequence position certain G R Wed May 02 18:48:31 BST 2018 This feature has been corrected as a result of our quality control procedures. The original label was 'gly1292arg' Tue Jun 30 16:40:03 BST 2020 This feature has been corrected as a result of our quality control procedures. The original label was 'p.Gly1292Arg' SV2B 5 western blot western blot Immuno blot Immuno blot 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 SV2A 4 western blot western blot Immuno blot Immuno blot 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 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 1 SV2B 5 western blot western blot Immuno blot Immuno blot 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 SV2A 4 western blot western blot Immuno blot Immuno blot 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 HcE 7 western blot western blot Immuno blot Immuno blot 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 tag tandem tag tandem tag undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined physical association physical association 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 1 HcF 6 western blot western blot Immuno blot Immuno blot 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 tag tandem tag tandem tag undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined SV2A 4 western blot western blot Immuno blot Immuno blot 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 SV2B 5 western blot western blot Immuno blot Immuno blot 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 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