| IED ID | IndEnz0002016398 |
| Enzyme Type ID | protease016398 |
| Protein Name |
Botulinum neurotoxin type F BoNT/F Bontoxilysin-F Cleaved into: Botulinum neurotoxin F light chain LC EC 3.4.24.69 ; Botulinum neurotoxin F heavy chain HC |
| Gene Name | F bont boNT/F CLI_0851 |
| Organism | Clostridium botulinum (strain Langeland / NCTC 10281 / Type F) |
| Taxonomic Lineage | cellular organisms Bacteria Terrabacteria group Firmicutes Clostridia Eubacteriales Clostridiaceae Clostridium Clostridium botulinum Clostridium botulinum F Clostridium botulinum (strain Langeland / NCTC 10281 / Type F) |
| Enzyme Sequence | MPVVINSFNYNDPVNDDTILYMQIPYEEKSKKYYKAFEIMRNVWIIPERNTIGTDPSDFDPPASLENGSSAYYDPNYLTTDAEKDRYLKTTIKLFKRINSNPAGEVLLQEISYAKPYLGNEHTPINEFHPVTRTTSVNIKSSTNVKSSIILNLLVLGAGPDIFENSSYPVRKLMDSGGVYDPSNDGFGSINIVTFSPEYEYTFNDISGGYNSSTESFIADPAISLAHELIHALHGLYGARGVTYKETIKVKQAPLMIAEKPIRLEEFLTFGGQDLNIITSAMKEKIYNNLLANYEKIATRLSRVNSAPPEYDINEYKDYFQWKYGLDKNADGSYTVNENKFNEIYKKLYSFTEIDLANKFKVKCRNTYFIKYGFLKVPNLLDDDIYTVSEGFNIGNLAVNNRGQNIKLNPKIIDSIPDKGLVEKIVKFCKSVIPRKGTKAPPRLCIRVNNRELFFVASESSYNENDINTPKEIDDTTNLNNNYRNNLDEVILDYNSETIPQISNQTLNTLVQDDSYVPRYDSNGTSEIEEHNVVDLNVFFYLHAQKVPEGETNISLTSSIDTALSEESQVYTFFSSEFINTINKPVHAALFISWINQVIRDFTTEATQKSTFDKIADISLVVPYVGLALNIGNEVQKENFKEAFELLGAGILLEFVPELLIPTILVFTIKSFIGSSENKNKIIKAINNSLMERETKWKEIYSWIVSNWLTRINTQFNKRKEQMYQALQNQVDAIKTVIEYKYNNYTSDERNRLESEYNINNIREELNKKVSLAMENIERFITESSIFYLMKLINEAKVSKLREYDEGVKEYLLDYISEHRSILGNSVQELNDLVTSTLNNSIPFELSSYTNDKILILYFNKLYKKIKDNSILDMRYENNKFIDISGYGSNISINGDVYIYSTNRNQFGIYSSKPSEVNIAQNNDIIYNGRYQNFSISFWVRIPKYFNKVNLNNEYTIIDCIRNNNSGWKISLNYNKIIWTLQDTAGNNQKLVFNYTQMISISDYINKWIFVTITNNRLGNSRIYINGNLIDEKSISNLGDIHVSDNILFKIVGCNDTRYVGIRYFKVFDTELGKTEIETLYSDEPDPSILKDFWGNYLLYNKRYYLLNLLRTDKSITQNSNFLNINQQRGVYQKPNIFSNTRLYTGVEVIIRKNGSTDISNTDNFVRKNDLAYINVVDRDVEYRLYADISIAKPEKIIKLIRTSNSNNSLGQIIVMDSIGNNCTMNFQNNNGGNIGLLGFHSNNLVASSWYYNNIRKNTSSNGCFWSFISKEHGWQEN |
| Enzyme Length | 1278 |
| Uniprot Accession Number | A7GBG3 |
| Absorption | |
| Active Site | ACT_SITE 228; /evidence=ECO:0000255|PROSITE-ProRule:PRU10095 |
| Activity Regulation | |
| Binding Site | BINDING 1111; /note="Host ganglioside GD1a binding"; /evidence="ECO:0000269|PubMed:21849494"; BINDING 1195; /note="Host ganglioside GD1a binding"; /evidence="ECO:0000269|PubMed:21849494, ECO:0007744|PDB:3RSJ"; BINDING 1256; /note="Host ganglioside GD1a binding"; /evidence="ECO:0000269|PubMed:21849494, ECO:0007744|PDB:3RSJ" |
| Calcium Binding | |
| catalytic Activity | CATALYTIC ACTIVITY: Reaction=Limited hydrolysis of proteins of the neuroexocytosis apparatus, synaptobrevins, SNAP25 or syntaxin. No detected action on small molecule substrates.; EC=3.4.24.69; Evidence={ECO:0000305|PubMed:19476346}; |
| DNA Binding | |
| EC Number | 3.4.24.69 |
| Enzyme Function | FUNCTION: [Botulinum neurotoxin type F]: Botulinum toxin causes flaccid paralysis by inhibiting neurotransmitter (acetylcholine) release from the presynaptic membranes of nerve terminals of the eukaryotic host skeletal and autonomic nervous system, with frequent heart or respiratory failure (PubMed:14423425). Precursor of botulinum neurotoxin F which may have 2 coreceptors; complex polysialylated gangliosides found on neural tissue and specific membrane-anchored proteins found in synaptic vesicles. Receptor proteins are exposed on host presynaptic cell membrane during neurotransmitter release, when the toxin heavy chain (HC) binds to them (PubMed:19476346, PubMed:19650874). Upon synaptic vesicle recycling the toxin is taken up via the endocytic pathway. When the pH of the toxin-containing endosome drops a structural rearrangement occurs so that the N-terminus of the HC forms pores that allows the light chain (LC) to translocate into the cytosol. Once in the cytosol the disulfide bond linking the 2 subunits is reduced and LC cleaves its target protein on synaptic vesicles, preventing their fusion with the cytoplasmic membrane and thus neurotransmitter release (By similarity). Requires complex gangliosides for full neurotoxicity (PubMed:19650874, PubMed:21483489). Electrical stimulation increases uptake of toxin, presumably by transiently exposing a receptor usually found in eukaryotic target synaptic vesicles (PubMed:19476346, PubMed:19650874). Blocks neurotransitter release by cleaving synaptobrevin-2/VAMP2 (PubMed:19476346). It is not clear whether a synaptic vesicle protein acts as its receptor; there is evidence for and against SV2 fulfilling this function (PubMed:19650874, PubMed:21483489, PubMed:19476346). {ECO:0000250|UniProtKB:P0DPI0, ECO:0000269|PubMed:14423425, ECO:0000269|PubMed:19476346, ECO:0000269|PubMed:19650874, ECO:0000269|PubMed:21483489}.; FUNCTION: [Botulinum neurotoxin F light chain]: Has protease activity (PubMed:19476346, PubMed:19543288). After translocation into the eukaryotic host cytosol, inhibits neurotransmitter release by acting as a zinc endopeptidase that catalyzes the hydrolysis of the '58-Gln-|-Lys-59' bond of synaptobrevin-2/VAMP2 and probably also the equivalent 'Gln-|-Lys' sites in VAMP1 and VAMP3 (PubMed:19476346, PubMed:19543288). Substrate specificity is conferred by multiple interactions of LC with substrate (PubMed:19543288). {ECO:0000250|UniProtKB:P30996, ECO:0000269|PubMed:19543288, ECO:0000305|PubMed:19476346}.; FUNCTION: [Botulinum neurotoxin F heavy chain]: Responsible for host epithelial cell transcytosis, host nerve cell targeting and translocation of light chain (LC) into host cytosol. Composed of 3 subdomains; the translocation domain (TD), and N-terminus and C-terminus of the receptor-binding domain (RBD). The RBD is responsible for the adherence of the toxin to the cell surface (PubMed:19476346, PubMed:19650874). The N-terminus of the TD wraps an extended belt around the perimeter of the LC, protecting Zn(2+) in the active site; it may also prevent premature LC dissociation from the translocation channel and protect toxin prior to translocation (By similarity). Isolated HC binds to host synaptosomes and neurons, significantly decreases uptake and toxicity of whole BoNT/F (PubMed:19476346, PubMed:19650874). Interferes with uptake of BoNT/E and to a lesser extent BoNT/C (PubMed:19650874). in vitro binds gangliosides GT1b, GD1b and GD1a (PubMed:19650874, PubMed:19476346, PubMed:21849494). Binds to synaptic vesicle glycoproteins SV2A, SV2B and SV2C which may serve as coreceptors with gangliosides (PubMed:19650874, PubMed:19476346). Interaction with SV2 proteins requires SV2 glycosylation (PubMed:19476346). However knockout SV2A/SV2B mice still cleave synaptobrevin, leaving the identification of its receptor unclear (PubMed:21483489). {ECO:0000250|UniProtKB:P0DPI0, ECO:0000269|PubMed:19476346, ECO:0000269|PubMed:19650874, ECO:0000269|PubMed:21483489}. |
| Temperature Dependency | |
| PH Dependency | |
| Pathway | |
| nucleotide Binding | |
| Features | Active site (1); Beta strand (47); Binding site (3); Chain (3); Coiled coil (1); Disulfide bond (1); Helix (19); Metal binding (3); Motif (1); Mutagenesis (13); Region (6); Turn (10) |
| Keywords | 3D-structure;Coiled coil;Disulfide bond;Host cell junction;Host cell membrane;Host cytoplasm;Host cytoplasmic vesicle;Host membrane;Host synapse;Hydrolase;Lipid-binding;Membrane;Metal-binding;Metalloprotease;Neurotoxin;Protease;Secreted;Toxin;Virulence;Zinc |
| Interact With | P63027 |
| Induction | |
| Subcellular Location | SUBCELLULAR LOCATION: [Botulinum neurotoxin type F]: Secreted {ECO:0000250|UniProtKB:P0DPI0}.; SUBCELLULAR LOCATION: [Botulinum neurotoxin F light chain]: Secreted {ECO:0000250|UniProtKB:P0DPI0}. Host cytoplasm, host cytosol {ECO:0000305|PubMed:19476346, ECO:0000305|PubMed:19543288}.; SUBCELLULAR LOCATION: [Botulinum neurotoxin F heavy chain]: Secreted {ECO:0000250|UniProtKB:P0DPI0}. Host cell junction, host synapse, host presynaptic cell membrane {ECO:0000250|UniProtKB:P0DPI0}. Host cytoplasmic vesicle, host secretory vesicle, host synaptic vesicle membrane {ECO:0000305|PubMed:19476346}; Multi-pass membrane protein {ECO:0000305}. |
| Modified Residue | |
| Post Translational Modification | |
| Signal Peptide | |
| Structure 3D | X-ray crystallography (4) |
| Cross Reference PDB | 3FIE; 3FII; 3FUQ; 3RSJ; |
| Mapped Pubmed ID | 22172278; |
| Motif | MOTIF 1245..1248; /note="Host ganglioside-binding motif"; /evidence="ECO:0000250|UniProtKB:P0DPI0, ECO:0000305|PubMed:19476346, ECO:0000305|PubMed:19650874" |
| Gene Encoded By | |
| Mass | 147,075 |
| Kinetics | BIOPHYSICOCHEMICAL PROPERTIES: Kinetic parameters: KM=19.0 uM for over-expressed human VAMP1 {ECO:0000269|PubMed:22289120}; KM=24.5 uM for over-expressed human VAMP2 {ECO:0000269|PubMed:22289120}; KM=15.0 uM for over-expressed human VAMP3 {ECO:0000269|PubMed:22289120}; Note=kcat is 16.12, 34.37 and 28.57 sec(-1) for over-expressed human VAMP1, VAMP2 and VAMP3 respectively. {ECO:0000269|PubMed:22289120}; |
| Metal Binding | METAL 227; /note="Zinc; via tele nitrogen; catalytic"; /evidence="ECO:0000255|PROSITE-ProRule:PRU10095, ECO:0000269|PubMed:19543288, ECO:0007744|PDB:3FIE, ECO:0007744|PDB:3FII"; METAL 231; /note="Zinc; via tele nitrogen; catalytic"; /evidence="ECO:0000255|PROSITE-ProRule:PRU10095, ECO:0000269|PubMed:19543288, ECO:0007744|PDB:3FIE, ECO:0007744|PDB:3FII"; METAL 266; /note="Zinc; catalytic"; /evidence="ECO:0000269|PubMed:19543288, ECO:0007744|PDB:3FIE, ECO:0007744|PDB:3FII" |
| Rhea ID | |
| Cross Reference Brenda | 3.4.24.69; |