| IED ID | IndEnz0002007650 |
| Enzyme Type ID | protease007650 |
| 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 | botF |
| Organism | Clostridium botulinum |
| Taxonomic Lineage | cellular organisms Bacteria Terrabacteria group Firmicutes Clostridia Eubacteriales Clostridiaceae Clostridium Clostridium botulinum |
| Enzyme Sequence | MPVAINSFNYNDPVNDDTILYMQIPYEEKSKKYYKAFEIMRNVWIIPERNTIGTNPSDFDPPASLKNGSSAYYDPNYLTTDAEKDRYLKTTIKLFKRINSNPAGKVLLQEISYAKPYLGNDHTPIDEFSPVTRTTSVNIKLSTNVESSMLLNLLVLGAGPDIFESCCYPVRKLIDPDVVYDPSNYGFGSINIVTFSPEYEYTFNDISGGHNSSTESFIADPAISLAHELIHALHGLYGARGVTYEETIEVKQAPLMIAEKPIRLEEFLTFGGQDLNIITSAMKEKIYNNLLANYEKIATRLSEVNSAPPEYDINEYKDYFQWKYGLDKNADGSYTVNENKFNEIYKKLYSFTESDLANKFKVKCRNTYFIKYEFLKVPNLLDDDIYTVSEGFNIGNLAVNNRGQSIKLNPKIIDSIPDKGLVEKIVKFCKSVIPRKGTKAPPRLCIRVNNSELFFVASESSYNENDINTPKEIDDTTNLNNNYRNNLDEVILDYNSQTIPQISNRTLNTLVQDNSYVPRYDSNGTSEIEEYDVVDFNVFFYLHAQKVPEGETNISLTSSIDTALLEESKDIFFSSEFIDTINKPVNAALFIDWISKVIRDFTTEATQKSTVDKIADISLIVPYVGLALNIIIEAEKGNFEEAFELLGVGILLEFVPELTIPVILVFTIKSYIDSYENKNKAIKAINNSLIEREAKWKEIYSWIVSNWLTRINTQFNKRKEQMYQALQNQVDAIKTAIEYKYNNYTSDEKNRLESEYNINNIEEELNKKVSLAMKNIERFMTESSISYLMKLINEAKVGKLKKYDNHVKSDLLNYILDHRSILGEQTNELSDLVTSTLNSSIPFELSSYTNDKILIIYFNRLYKKIKDSSILDMRYENNKFIDISGYGSNISINGNVYIYSTNRNQFGIYNSRLSEVNIAQNNDIIYNSRYQNFSISFWVRIPKHYKPMNHNREYTIINCMGNNNSGWKISLRTVRDCEIIWTLQDTSGNKENLIFRYEELNRISNYINKWIFVTITNNRLGNSRIYINGNLIVEKSISNLGDIHVSDNILFKIVGCDDETYVGIRYFKVFNTELDKTEIETLYSNEPDPSILKNYWGNYLLYNKKYYLFNLLRKDKYITLNSGILNINQQRGVTEGSVFLNYKLYEGVEVIIRKNGPIDISNTDNFVRKNDLAYINVVDRGVEYRLYADTKSEKEKIIRTSNLNDSLGQIIVMDSIGNNCTMNFQNNNGSNIGLLGFHSNNLVASSWYYNNIRRNTSSNGCFWSSISKENGWKE |
| Enzyme Length | 1274 |
| Uniprot Accession Number | P30996 |
| Absorption | |
| Active Site | ACT_SITE 228; /evidence=ECO:0000255|PROSITE-ProRule:PRU10095 |
| Activity Regulation | ACTIVITY REGULATION: Proteolysis inhibited by 1,10-phenanthroline, captopril and EDTA (PubMed:8505288). {ECO:0000269|PubMed:8505288}. |
| Binding Site | |
| 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:0000269|PubMed:8197120, ECO:0000269|PubMed:8505288}; |
| 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. 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. 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). Whole toxin only has protease activity after reduction, which releases LC (PubMed:8505288). Requires complex eukaryotic host polysialogangliosides for full neurotoxicity (By similarity). It is not clear whether a synaptic vesicle protein acts as its receptor; there is evidence for and against SV2 fulfilling this function (By similarity). {ECO:0000250|UniProtKB:A7GBG3, ECO:0000269|PubMed:8505288}.; FUNCTION: [Botulinum neurotoxin F light chain]: Has proteolytic activity (PubMed:8505288, PubMed:8175689, PubMed:8197120). After translocation into the eukaryotic host cytosol, inhibits neurotransmitter release by acting as a zinc endopeptidase that catalyzes the hydrolysis of the '60-Gln-|-Lys-61' bond of synaptobrevin-1/VAMP1 and the equivalent 'Gln-|-Lys' sites in VAMP2 and VAMP3 (PubMed:8505288, PubMed:8175689). Cleaves the '48-Gln-|-Lys-49' bond of A.californica synaptobrevin (AC P35589) (PubMed:8197120). {ECO:0000269|PubMed:8175689, ECO:0000269|PubMed:8197120, ECO:0000269|PubMed:8505288, ECO:0000305|PubMed:16128577}.; 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. It simultaneously recognizes 2 coreceptors; polysialated gangliosides and the receptor protein SV2A, SV2B and SV2C in close proximity on host synaptic vesicles; although not all evidence indicates these are the receptors (By similarity). 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). The TD inserts into synaptic vesicle membrane to allow translocation into the host cytosol (By similarity). {ECO:0000250|UniProtKB:A7GBG3}. |
| Temperature Dependency | |
| PH Dependency | |
| Pathway | |
| nucleotide Binding | |
| Features | Active site (1); Beta strand (18); Chain (3); Disulfide bond (1); Helix (14); Metal binding (3); Motif (1); Region (4); Turn (7) |
| Keywords | 3D-structure;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;Transmembrane;Virulence;Zinc |
| Interact With | |
| 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:8175689, ECO:0000305|PubMed:8505288}.; SUBCELLULAR LOCATION: [Botulinum neurotoxin F heavy chain]: Secreted {ECO:0000250|UniProtKB:P0DPI0}. Host cell junction, host synapse, host presynaptic cell membrane {ECO:0000305}. Host cytoplasmic vesicle, host secretory vesicle, host synaptic vesicle membrane {ECO:0000250|UniProtKB:P0DPI0}; Multi-pass membrane protein {ECO:0000305}. |
| Modified Residue | |
| Post Translational Modification | |
| Signal Peptide | |
| Structure 3D | X-ray crystallography (2) |
| Cross Reference PDB | 2A8A; 2A97; |
| Mapped Pubmed ID | 24583011; |
| Motif | MOTIF 1245..1248; /note=Host ganglioside-binding motif; /evidence=ECO:0000250|UniProtKB:P0DPI0 |
| Gene Encoded By | |
| Mass | 146,710 |
| Kinetics | |
| Metal Binding | METAL 227; /note="Zinc; via tele nitrogen; catalytic"; /evidence="ECO:0000255|PROSITE-ProRule:PRU10095, ECO:0000269|PubMed:16128577, ECO:0007744|PDB:2A8A, ECO:0007744|PDB:2A97"; METAL 231; /note="Zinc; via tele nitrogen; catalytic"; /evidence="ECO:0000255|PROSITE-ProRule:PRU10095, ECO:0000269|PubMed:16128577, ECO:0007744|PDB:2A8A, ECO:0007744|PDB:2A97"; METAL 266; /note="Zinc; catalytic"; /evidence="ECO:0000269|PubMed:16128577, ECO:0007744|PDB:2A8A, ECO:0007744|PDB:2A97" |
| Rhea ID | |
| Cross Reference Brenda | 3.4.24.69; |