UniProt functional annotation for P30996



	UniProt functional annotation for P30996

UniProt code: P30996.

Organism: Clostridium botulinum.

Taxonomy: Bacteria; Firmicutes; Clostridia; Eubacteriales; Clostridiaceae; Clostridium.

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}.

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};

Cofactor: Name=Zn(2+); Xref=ChEBI:CHEBI:29105; Evidence={ECO:0000269|PubMed:16128577, ECO:0000305|PubMed:8505288}; Note=Binds 1 zinc ion per subunit (PubMed:8505288, PubMed:16128577). {ECO:0000269|PubMed:16128577, ECO:0000305|PubMed:8505288};

Activity regulation: Proteolysis inhibited by 1,10-phenanthroline, captopril and EDTA (PubMed:8505288). {ECO:0000269|PubMed:8505288}.

Subunit: Heterodimer; disulfide-linked heterodimer of a light chain (LC) and a heavy chain (HC). The LC has the proteolytic/pharmacological activity, while the N- and C-terminal of the HC mediate channel formation and toxin binding, respectively. Interacts with host synaptic vesicle glycoproteins SV2A, SV2B and SV2C (By similarity). {ECO:0000250|UniProtKB:A7GBG3}.

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}.

Domain: [Botulinum neurotoxin F light chain]: Has protease activity (PubMed:8505288, PubMed:8175689, PubMed:8197120). {ECO:0000269|PubMed:8175689, ECO:0000269|PubMed:8197120, ECO:0000269|PubMed:8505288}.

Domain: [Botulinum neurotoxin F heavy chain]: Has 3 functional domains; the translocation domain (TD) and the receptor-binding domain (RBD) which is further subdivided into N- and C-terminal domains (N-RBD and C-RBD) (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 and may be a pseudosubstrate inhibitor which serves as an intramolecular chaperone for the LC prior to its translocation into the host cytosol (By similarity). The RBD binds transiently exposed coreceptors on the host presynaptic cell membrane (By similarity). {ECO:0000250|UniProtKB:A7GBG3}.

Miscellaneous: There are seven antigenically distinct forms of botulinum neurotoxin: Types A, B, C, D, E, F, and G; new subtypes are quite frequent.

Miscellaneous: Botulism poisoning is usually food-borne, either by ingesting toxin or bacterial-contaminated food, or less frequently by inhalation poisoning. In both cases the neurotoxin binds to the apical surface of epithelial cells in the gut or airway. Toxin undergoes receptor-mediated endocytosis (using a different receptor than on target nerve cells), transcytosis across the epithelial cells and release into the general circulation. Once in the general circulation it binds to its target cells. {ECO:0000250|UniProtKB:P0DPI0}.

Similarity: Belongs to the peptidase M27 family. {ECO:0000305}.

Annotations taken from UniProtKB at the EBI.


Organism:		Clostridium botulinum.
Taxonomy:		Bacteria; Firmicutes; Clostridia; Eubacteriales; Clostridiaceae; Clostridium.



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}.


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};
Cofactor:		Name=Zn(2+); Xref=ChEBI:CHEBI:29105; Evidence={ECO:0000269\|PubMed:16128577, ECO:0000305\|PubMed:8505288}; Note=Binds 1 zinc ion per subunit (PubMed:8505288, PubMed:16128577). {ECO:0000269\|PubMed:16128577, ECO:0000305\|PubMed:8505288};
Activity regulation:		Proteolysis inhibited by 1,10-phenanthroline, captopril and EDTA (PubMed:8505288). {ECO:0000269\|PubMed:8505288}.
Subunit:		Heterodimer; disulfide-linked heterodimer of a light chain (LC) and a heavy chain (HC). The LC has the proteolytic/pharmacological activity, while the N- and C-terminal of the HC mediate channel formation and toxin binding, respectively. Interacts with host synaptic vesicle glycoproteins SV2A, SV2B and SV2C (By similarity). {ECO:0000250\|UniProtKB:A7GBG3}.
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}.
Domain:		[Botulinum neurotoxin F light chain]: Has protease activity (PubMed:8505288, PubMed:8175689, PubMed:8197120). {ECO:0000269\|PubMed:8175689, ECO:0000269\|PubMed:8197120, ECO:0000269\|PubMed:8505288}.
Domain:		[Botulinum neurotoxin F heavy chain]: Has 3 functional domains; the translocation domain (TD) and the receptor-binding domain (RBD) which is further subdivided into N- and C-terminal domains (N-RBD and C-RBD) (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 and may be a pseudosubstrate inhibitor which serves as an intramolecular chaperone for the LC prior to its translocation into the host cytosol (By similarity). The RBD binds transiently exposed coreceptors on the host presynaptic cell membrane (By similarity). {ECO:0000250\|UniProtKB:A7GBG3}.
Miscellaneous:		There are seven antigenically distinct forms of botulinum neurotoxin: Types A, B, C, D, E, F, and G; new subtypes are quite frequent.
Miscellaneous:		Botulism poisoning is usually food-borne, either by ingesting toxin or bacterial-contaminated food, or less frequently by inhalation poisoning. In both cases the neurotoxin binds to the apical surface of epithelial cells in the gut or airway. Toxin undergoes receptor-mediated endocytosis (using a different receptor than on target nerve cells), transcytosis across the epithelial cells and release into the general circulation. Once in the general circulation it binds to its target cells. {ECO:0000250\|UniProtKB:P0DPI0}.
Similarity:		Belongs to the peptidase M27 family. {ECO:0000305}.