DIP Database of Interacting Proteins 611 Young Drive East; Los Angeles CA 90095; USA http://dip.doe-mbi.ucla.edu/ dip@mbi.ucla.edu Structural basis for recognition of synaptic vesicle protein 2C by botulinum neurotoxin A. Structural basis for recognition of synaptic vesicle protein 2C by botulinum neurotoxin A. Nature 2014 imex curation Benoit RM., Frey D., Hilbert M., Kevenaar JT., Wieser MM., Stirnimann CU., McMillan D., Ceska T., Lebon F., Jaussi R., Steinmetz MO., Schertler GF., Hoogenraad CC., Capitani G., Kammerer RA. Only protein-protein interactions in vitro In vitro pull down pull down weight by comassie molecular weight estimation by coomasie staining weight by comassie Benoit RM., Frey D., Hilbert M., Kevenaar JT., Wieser MM., Stirnimann CU., McMillan D., Ceska T., Lebon F., Jaussi R., Steinmetz MO., Schertler GF., Hoogenraad CC., Capitani G., Kammerer RA. DIP original experiment: DIP-259817X Nature 2014 IMEx Structural basis for recognition of synaptic vesicle protein 2C by botulinum neurotoxin A. Structural basis for recognition of synaptic vesicle protein 2C by botulinum neurotoxin A. Structural basis for recognition of synaptic vesicle protein 2C by botulinum neurotoxin A. Nature 2014 imex curation Benoit RM., Frey D., Hilbert M., Kevenaar JT., Wieser MM., Stirnimann CU., McMillan D., Ceska T., Lebon F., Jaussi R., Steinmetz MO., Schertler GF., Hoogenraad CC., Capitani G., Kammerer RA. Only protein-protein interactions in vitro In vitro x-ray diffraction x-ray crystallography X-ray X-ray x-ray diffraction predetermined predetermined participant predetermined Benoit RM., Frey D., Hilbert M., Kevenaar JT., Wieser MM., Stirnimann CU., McMillan D., Ceska T., Lebon F., Jaussi R., Steinmetz MO., Schertler GF., Hoogenraad CC., Capitani G., Kammerer RA. DIP original experiment: DIP-259815X Nature 2014 IMEx Structural basis for recognition of synaptic vesicle protein 2C by botulinum neurotoxin A. Structural basis for recognition of synaptic vesicle protein 2C by botulinum neurotoxin A. Structural basis for recognition of synaptic vesicle protein 2C by botulinum neurotoxin A. Nature 2014 imex curation Benoit RM., Frey D., Hilbert M., Kevenaar JT., Wieser MM., Stirnimann CU., McMillan D., Ceska T., Lebon F., Jaussi R., Steinmetz MO., Schertler GF., Hoogenraad CC., Capitani G., Kammerer RA. Only protein-protein interactions in vitro In vitro pull down pull down weight by comassie molecular weight estimation by coomasie staining weight by comassie Benoit RM., Frey D., Hilbert M., Kevenaar JT., Wieser MM., Stirnimann CU., McMillan D., Ceska T., Lebon F., Jaussi R., Steinmetz MO., Schertler GF., Hoogenraad CC., Capitani G., Kammerer RA. DIP original experiment: DIP-259818X Nature 2014 IMEx Structural basis for recognition of synaptic vesicle protein 2C by botulinum neurotoxin A. Structural basis for recognition of synaptic vesicle protein 2C by botulinum neurotoxin A. Structural basis for recognition of synaptic vesicle protein 2C by botulinum neurotoxin A. Nature 2014 imex curation Benoit RM., Frey D., Hilbert M., Kevenaar JT., Wieser MM., Stirnimann CU., McMillan D., Ceska T., Lebon F., Jaussi R., Steinmetz MO., Schertler GF., Hoogenraad CC., Capitani G., Kammerer RA. Only protein-protein interactions in vitro In vitro fluorescence spectr classical fluorescence spectroscopy fluorescence spectr predetermined predetermined participant predetermined Benoit RM., Frey D., Hilbert M., Kevenaar JT., Wieser MM., Stirnimann CU., McMillan D., Ceska T., Lebon F., Jaussi R., Steinmetz MO., Schertler GF., Hoogenraad CC., Capitani G., Kammerer RA. DIP original experiment: DIP-259816X Nature 2014 IMEx Structural basis for recognition of synaptic vesicle protein 2C by botulinum neurotoxin A. sv2c_human Synaptic vesicle glycoprotein 2C SV2C KIAA1054 protein protein human Homo sapiens Human MEDSYKDRTSLMKGAKDIAREVKKQTVKKVNQAVDRAQDEYTQRSYSRFQDEEDDDDYYPAGETYNGEANDDEGSSEATEGHDEDDEIYEGEYQGIPSMNQAKDSIVSVGQPKGDEYKDRRELESERRADEEELAQQYELIIQECGHGRFQWALFFVLGMALMADGVEVFVVGFVLPSAETDLCIPNSGSGWLGSIVYLGMMVGAFFWGGLADKVGRKQSLLICMSVNGFFAFLSSFVQGYGFFLFCRLLSGFGIGGAIPTVFSYFAEVLAREKRGEHLSWLCMFWMIGGIYASAMAWAIIPHYGWSFSMGSAYQFHSWRVFVIVCALPCVSSVVALTFMPESPRFLLEVGKHDEAWMILKLIHDTNMRARGQPEKVFTVNKIKTPKQIDELIEIESDTGTWYRRCFVRIRTELYGIWLTFMRCFNYPVRDNTIKLTIVWFTLSFGYYGLSVWFPDVIKPLQSDEYALLTRNVERDKYANFTINFTMENQIHTGMEYDNGRFIGVKFKSVTFKDSVFKSCTFEDVTSVNTYFKNCTFIDTVFDNTDFEPYKFIDSEFKNCSFFHNKTGCQITFDDDYSAYWIYFVNFLGTLAVLPGNIVSALLMDRIGRLTMLGGSMVLSGISCFFLWFGTSESMMIGMLCLYNGLTISAWNSLDVVTVELYPTDRRATGFGFLNALCKAAAVLGNLIFGSLVSITKSIPILLASTVLVCGGLVGLCLPDTRTQVLM DIP protein Q496J9 original sequence version: 79 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 bota-sv2c-1 2 6 unspecified role unspecified role unspecified role unspecified role Region 871-1296 Region 871-1296 sufficient to bind sufficient binding region sufficient to bind certain certain sequence position certain certain certain sequence position certain ecolx Escherichia coli 5 unspecified role unspecified role unspecified role unspecified role Region 456-574 Region 456-574 sufficient to bind sufficient binding region sufficient to bind certain certain sequence position certain certain certain sequence position certain ecolx Escherichia coli direct interaction direct interaction bota-sv2c-2 3 6 unspecified role unspecified role bait bait His-tag His-tag his tag his tag Hexa-His-tag 6-His-tag Histidine-tag Hexa-His-tag Histidine-tag 6-His-tag n-term range n-terminal range n-term range n-term range n-terminal range n-term range Region 871-1296 Region 871-1296 sufficient to bind sufficient binding region sufficient to bind certain certain sequence position certain certain certain sequence position certain ecolx Escherichia coli 5 unspecified role unspecified role prey prey Region 456-574 Region 456-574 sufficient to bind sufficient binding region sufficient to bind certain certain sequence position certain certain certain sequence position certain Q496J9:p.Phe563Ala F563A mutation disrupting strength mutation disrupting interaction strength mutation disrupting strength alanine scanning alanine scanning certain certain sequence position certain certain certain sequence position certain F A This feature has been corrected as a result of our quality control procedures. The original label was 'F563A'. Wed May 02 16:57:54 BST 2018 This feature has been corrected as a result of our quality control procedures. The original label was 'phe563ala' Tue Jun 30 16:51:40 BST 2020 This feature has been corrected as a result of our quality control procedures. The original label was 'p.Phe563Ala' ecolx Escherichia coli direct interaction direct interaction bota-sv2c-3 1 6 unspecified role unspecified role bait bait Region 871-1296 Region 871-1296 sufficient to bind sufficient binding region sufficient to bind certain certain sequence position certain certain certain sequence position certain P0DPI0:p.Arg1294Ala R1294A mutation decreasing strength mutation decreasing interaction strength mutation decreasing strength alanine scanning alanine scanning certain certain sequence position certain certain certain sequence position certain R A This feature has been corrected as a result of our quality control procedures. The original label was 'R1294A'. Wed May 02 19:26:28 BST 2018 This feature has been corrected as a result of our quality control procedures. The original label was 'arg1294ala' Tue Jun 30 19:54:37 BST 2020 This feature has been corrected as a result of our quality control procedures. The original label was 'p.Arg1294Ala' P0DPI0:p.Arg1156Glu R1156E mutation decreasing strength mutation decreasing interaction strength mutation decreasing strength substitut analysis substitution analysis substitut analysis certain certain sequence position certain certain certain sequence position certain R E This feature has been corrected as a result of our quality control procedures. The original label was 'R1156E'. Wed May 02 15:31:04 BST 2018 This feature has been corrected as a result of our quality control procedures. The original label was 'arg1156glu' Tue Jun 30 16:33:09 BST 2020 This feature has been corrected as a result of our quality control procedures. The original label was 'p.Arg1156Glu' P0DPI0:p.[Thr1145Ala;Thr1146Ala] T1145A/T1146A mutation disrupting strength mutation disrupting interaction strength mutation disrupting strength alanine scanning alanine scanning certain certain sequence position certain certain certain sequence position certain T A certain certain sequence position certain certain certain sequence position certain T A This feature has been corrected as a result of our quality control procedures. The original label was 'T1145A/T1146A'. Wed May 02 20:09:18 BST 2018 This feature has been corrected as a result of our quality control procedures. The original label was 'thr1145ala,thr1146ala' Tue Jun 30 17:03:08 BST 2020 This feature has been corrected as a result of our quality control procedures. The original label was 'p.[Thr1145Ala;Thr1146Ala]' His-tag His-tag his tag his tag Hexa-His-tag 6-His-tag Histidine-tag Hexa-His-tag Histidine-tag 6-His-tag n-term range n-terminal range n-term range n-term range n-terminal range n-term range ecolx Escherichia coli 5 unspecified role unspecified role prey prey Region 456-574 Region 456-574 sufficient to bind sufficient binding region sufficient to bind certain certain sequence position certain certain certain sequence position certain ecolx Escherichia coli direct interaction direct interaction bota-sv2c-4 4 6 unspecified role unspecified role unspecified role unspecified role Region 871-1296 Region 871-1296 sufficient to bind sufficient binding region sufficient to bind certain certain sequence position certain certain certain sequence position certain P0DPI0:p.Arg1294Ala R1294A mutation decreasing strength mutation decreasing interaction strength mutation decreasing strength alanine scanning alanine scanning certain certain sequence position certain certain certain sequence position certain R A This feature has been corrected as a result of our quality control procedures. The original label was 'R1294A'. Wed May 02 18:41:45 BST 2018 This feature has been corrected as a result of our quality control procedures. The original label was 'arg1294ala' Tue Jun 30 15:55:47 BST 2020 This feature has been corrected as a result of our quality control procedures. The original label was 'p.Arg1294Ala' P0DPI0:p.Arg1156Glu R1156E mutation decreasing strength mutation decreasing interaction strength mutation decreasing strength substitut analysis substitution analysis substitut analysis certain certain sequence position certain certain certain sequence position certain R E This feature has been corrected as a result of our quality control procedures. The original label was 'R1156E'. Wed May 02 14:36:08 BST 2018 This feature has been corrected as a result of our quality control procedures. The original label was 'arg1156glu' Tue Jun 30 20:13:46 BST 2020 This feature has been corrected as a result of our quality control procedures. The original label was 'p.Arg1156Glu' P0DPI0:p.[Thr1145Ala;Thr1146Ala] T1145A/T1146A mutation decreasing strength mutation decreasing interaction strength mutation decreasing strength alanine scanning alanine scanning certain certain sequence position certain certain certain sequence position certain T A certain certain sequence position certain certain certain sequence position certain T A This feature has been corrected as a result of our quality control procedures. The original label was 'T1145A/T1146A'. Wed May 02 20:28:28 BST 2018 This feature has been corrected as a result of our quality control procedures. The original label was 'thr1145ala,thr1146ala' Tue Jun 30 16:51:47 BST 2020 This feature has been corrected as a result of our quality control procedures. The original label was 'p.[Thr1145Ala;Thr1146Ala]' ecolx Escherichia coli 5 unspecified role unspecified role unspecified role unspecified role Region 456-574 Region 456-574 sufficient to bind sufficient binding region sufficient to bind certain certain sequence position certain certain certain sequence position certain Fluorescein-label Fluorescein-label fluor-5-maleimide fluorescein-5-maleimide label fluor-5-maleimide undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined ecolx Escherichia coli direct interaction direct interaction