| IED ID | IndEnz0002014557 |
| Enzyme Type ID | protease014557 |
| Protein Name |
Botulinum neurotoxin type G BoNT/G Bontoxilysin-G Cleaved into: Botulinum neurotoxin G light chain LC EC 3.4.24.69 ; Botulinum neurotoxin G heavy chain HC |
| Gene Name | botG |
| Organism | Clostridium botulinum |
| Taxonomic Lineage | cellular organisms Bacteria Terrabacteria group Firmicutes Clostridia Eubacteriales Clostridiaceae Clostridium Clostridium botulinum |
| Enzyme Sequence | MPVNIKXFNYNDPINNDDIIMMEPFNDPGPGTYYKAFRIIDRIWIVPERFTYGFQPDQFNASTGVFSKDVYEYYDPTYLKTDAEKDKFLKTMIKLFNRINSKPSGQRLLDMIVDAIPYLGNASTPPDKFAANVANVSINKKIIQPGAEDQIKGLMTNLIIFGPGPVLSDNFTDSMIMNGHSPISEGFGARMMIRFCPSCLNVFNNVQENKDTSIFSRRAYFADPALTLMHELIHVLHGLYGIKISNLPITPNTKEFFMQHSDPVQAEELYTFGGHDPSVISPSTDMNIYNKALQNFQDIANRLNIVSSAQGSGIDISLYKQIYKNKYDFVEDPNGKYSVDKDKFDKLYKALMFGFTETNLAGEYGIKTRYSYFSEYLPPIKTEKLLDNTIYTQNEGFNIASKNLKTEFNGQNKAVNKEAYEEISLEHLVIYRIAMCKPVMYKNTGKSEQCIIVNNEDLFFIANKDSFSKDLAKAETIAYNTQNNTIENNFSIDQLILDNDLSSGIDLPNENTEPFTNFDDIDIPVYIKQSALKKIFVDGDSLFEYLHAQTFPSNIENLQLTNSLNDALRNNNKVYTFFSTNLVEKANTVVGASLFVNWVKGVIDDFTSESTQKSTIDKVSDVSIIIPYIGPALNVGNETAKENFKNAFEIGGAAILMEFIPELIVPIVGFFTLESYVGNKGHIIMTISNALKKRDQKWTDMYGLIVSQWLSTVNTQFYTIKERMYNALNNQSQAIEKIIEDQYNRYSEEDKMNINIDFNDIDFKLNQSINLAINNIDDFINQCSISYLMNRMIPLAVKKLKDFDDNLKRDLLEYIDTNELYLLDEVNILKSKVNRHLKDSIPFDLSLYTKDTILIQVFNNYISNISSNAILSLSYRGGRLIDSSGYGATMNVGSDVIFNDIGNGQFKLNNSENSNITAHQSKFVVYDSMFDNFSINFWVRTPKYNNNDIQTYLQNEYTIISCIKNDSGWKVSIKGNRIIWTLIDVNAKSKSIFFEYSIKDNISDYINKWFSITITNDRLGNANIYINGSLKKSEKILNLDRINSSNDIDFKLINCTDTTKFVWIKDFNIFGRELNATEVSSLYWIQSSTNTLKDFWGNPLRYDTQYYLFNQGMQNIYIKYFSKASMGETAPRTNFNNAAINYQNLYLGLRFIIKKASNSRNINNDNIVREGDYIYLNIDNISDESYRVYVLVNSKEIQTQLFLAPINDDPTFYDVLQIKKYYEKTTYNCQILCEKDTKTFGLFGIGKFVKDYGYVWDTYDNYFCISQWYLRRISENINKLRLGCNWQFIPVDEGWTE |
| Enzyme Length | 1297 |
| Uniprot Accession Number | Q60393 |
| Absorption | |
| Active Site | ACT_SITE 231; /evidence=ECO:0000255|PROSITE-ProRule:PRU10095 |
| Activity Regulation | |
| 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:7910017}; |
| DNA Binding | |
| EC Number | 3.4.24.69 |
| Enzyme Function | FUNCTION: [Botulinum neurotoxin type G]: 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:15123599). Precursor of botulinum neurotoxin G which has 2 coreceptors; complex polysialylated gangliosides found on neural tissue and specific membrane-anchored proteins found in synaptic vesicles (PubMed:15123599). 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). Binds to host peripheral neuronal presynaptic membranes via synaptotagmins 1 and 2 (SYT1 and SYT2) (PubMed:15123599). Toxin binds to the membrane proximal extra-cytoplasmic region of SYT1 and SYT2 that is transiently exposed outside of cells during exocytosis; exogenous gangliosides do not enhance binding and subsequent uptake of toxin into host cells (PubMed:15123599). Toxin activity can be blocked by the appropriate synaptotagmin protein fragments in cell culture (PubMed:15123599). {ECO:0000250|UniProtKB:P0DPI0, ECO:0000269|PubMed:15123599}.; FUNCTION: [Botulinum neurotoxin G light chain]: Has proteolytic activity. After translocation into the eukaryotic host cytosol acts as a zinc endopeptidase that cleaves synaptobrevins-1, -2 and -3 (also called VAMP1, VAMP2 and VAMP3) (PubMed:7910017). Hydrolyzes the '81-Ala-|-Ala-82' bond of VAMP2, and probably also the equivalent 'Ala-|-Ala' sites in VAMP1 and VAMP3 (PubMed:7910017). Has low activity on A.californica synaptobrevin and none on D.melanogaster synaptobrevin or cellubrevin, have a slightly different sequence (PubMed:7910017). {ECO:0000269|PubMed:7910017}.; FUNCTION: [Botulinum neurotoxin G heavy chain]: Responsible for host 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 (N-RBD, C-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 proteins SYT1 and SYT2 in close proximity on host synaptic vesicles (PubMed:17185412). 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). Has 2 coreceptors; complex gangliosides found primarily on neural tissue and host synaptotagmin-1 and -2 (SYT1 and SYT2) which bind to separate sites at the tip of the HC (PubMed:17185412). HC alone binds to host receptors SYT1 and SYT2; C-RBD interacts with host SYT2 (PubMed:15123599). Has equal affinity for SYT1 and SYT2; gangliosides are not required for (or only very slightly improve) binding to a membrane-anchored receptor fragment (PubMed:17185412, PubMed:20219474). Has also been shown to only bind SYT1; the assay methods were different (PubMed:20507178). Binds ganglioside GT1b (PubMed:20507178). Significantly decreases uptake and toxicity of whole BoNT/B and BoNT/G (PubMed:19650874). {ECO:0000250|UniProtKB:P0DPI0, ECO:0000269|PubMed:15123599, ECO:0000269|PubMed:17185412, ECO:0000269|PubMed:19650874, ECO:0000269|PubMed:20219474, ECO:0000269|PubMed:20507178}. |
| Temperature Dependency | |
| PH Dependency | |
| Pathway | |
| nucleotide Binding | |
| Features | Active site (1); Beta strand (51); Chain (3); Disulfide bond (1); Helix (17); Initiator methionine (1); Metal binding (3); Motif (1); Mutagenesis (7); Region (4); Site (1); Turn (8) |
| 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;Virulence;Zinc |
| Interact With | |
| Induction | |
| Subcellular Location | SUBCELLULAR LOCATION: [Botulinum neurotoxin type G]: Secreted {ECO:0000269|PubMed:74236, ECO:0000305|PubMed:4922309}.; SUBCELLULAR LOCATION: [Botulinum neurotoxin G light chain]: Secreted {ECO:0000305|PubMed:74236}. Host cytoplasm, host cytosol {ECO:0000305|PubMed:7910017}.; SUBCELLULAR LOCATION: [Botulinum neurotoxin G heavy chain]: Secreted {ECO:0000305|PubMed:74236}. Host cell junction, host synapse, host presynaptic cell membrane {ECO:0000250|UniProtKB:P0DPI0}. 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 (3) |
| Cross Reference PDB | 1ZB7; 2VXR; 3MPP; |
| Mapped Pubmed ID | 22265973; |
| Motif | MOTIF 1266..1269; /note=Host ganglioside-binding motif; /evidence=ECO:0000305|PubMed:17185412 |
| Gene Encoded By | |
| Mass | 149,146 |
| Kinetics | |
| Metal Binding | METAL 230; /note="Zinc; via tele nitrogen; catalytic"; /evidence="ECO:0000255|PROSITE-ProRule:PRU10095, ECO:0000269|PubMed:16008342, ECO:0007744|PDB:1ZB7"; METAL 234; /note="Zinc; via tele nitrogen; catalytic"; /evidence="ECO:0000255|PROSITE-ProRule:PRU10095, ECO:0000269|PubMed:16008342, ECO:0007744|PDB:1ZB7"; METAL 268; /note="Zinc; catalytic"; /evidence="ECO:0000269|PubMed:16008342, ECO:0007744|PDB:1ZB7" |
| Rhea ID | |
| Cross Reference Brenda | 3.4.24.69; |