BXF_CLOBL
ID BXF_CLOBL Reviewed; 1278 AA.
AC A7GBG3; Q79AH9;
DT 18-JUL-2018, integrated into UniProtKB/Swiss-Prot.
DT 11-SEP-2007, sequence version 1.
DT 03-AUG-2022, entry version 111.
DE RecName: Full=Botulinum neurotoxin type F;
DE Short=BoNT/F {ECO:0000303|Ref.1};
DE AltName: Full=Bontoxilysin-F;
DE Contains:
DE RecName: Full=Botulinum neurotoxin F light chain;
DE Short=LC;
DE EC=3.4.24.69;
DE Contains:
DE RecName: Full=Botulinum neurotoxin F heavy chain;
DE Short=HC;
DE Flags: Precursor;
GN Name=F {ECO:0000312|EMBL:ABS41202.1};
GN Synonyms=bont {ECO:0000312|EMBL:CAA57358.1},
GN boNT/F {ECO:0000312|EMBL:ABS41202.1};
GN OrderedLocusNames=CLI_0851 {ECO:0000312|EMBL:ABS41202.1};
OS Clostridium botulinum (strain Langeland / NCTC 10281 / Type F).
OC Bacteria; Firmicutes; Clostridia; Eubacteriales; Clostridiaceae;
OC Clostridium.
OX NCBI_TaxID=441772;
RN [1]
RP NUCLEOTIDE SEQUENCE [GENOMIC DNA].
RC STRAIN=Langeland / NCTC 10281 / Type F;
RA Hutson R.A., Collins M.D.;
RT "The sequence of the gene encoding type F neurotoxin of clostridium
RT botulinum NCTC 10281; Comparative analysis with other botulinal
RT neurotoxins.";
RL Submitted (SEP-1994) to the EMBL/GenBank/DDBJ databases.
RN [2]
RP NUCLEOTIDE SEQUENCE [GENOMIC DNA].
RC STRAIN=Langeland / NCTC 10281 / Type F;
RX PubMed=20511432; DOI=10.1128/aem.03109-09;
RA Raphael B.H., Choudoir M.J., Luquez C., Fernandez R., Maslanka S.E.;
RT "Sequence diversity of genes encoding botulinum neurotoxin type F.";
RL Appl. Environ. Microbiol. 76:4805-4812(2010).
RN [3]
RP NUCLEOTIDE SEQUENCE [LARGE SCALE GENOMIC DNA].
RC STRAIN=Langeland / NCTC 10281 / Type F;
RA Brinkac L.M., Daugherty S., Dodson R.J., Madupu R., Brown J.L., Bruce D.,
RA Detter C., Munk C., Smith L.A., Smith T.J., White O., Brettin T.S.;
RL Submitted (JUN-2007) to the EMBL/GenBank/DDBJ databases.
RN [4]
RP NUCLEOTIDE SEQUENCE [GENOMIC DNA] OF 635-1000.
RC STRAIN=Langeland / NCTC 10281 / Type F;
RX PubMed=8408542; DOI=10.1128/jcm.31.9.2255-2262.1993;
RA Campbell K.D., Collins M.D., East A.K.;
RT "Gene probes for identification of the botulinal neurotoxin gene and
RT specific identification of neurotoxin types B, E, and F.";
RL J. Clin. Microbiol. 31:2255-2262(1993).
RN [5]
RP IDENTIFICATION, AND FUNCTION.
RX PubMed=14423425; DOI=10.1111/j.1699-0463.1960.tb04741.x;
RA Moller V., Scheibel I.;
RT "Preliminary report on the isolation of an apparently new type of Cl.
RT botulinum.";
RL Acta Pathol. Microbiol. Scand. 48:80-80(1960).
RN [6]
RP FUNCTION (BOTULINUM NEUROTOXIN TYPE F), FUNCTION (BOTULINUM NEUROTOXIN F
RP HEAVY CHAIN), INTERACTION WITH HOST SV2 PROTEINS, GANGLIOSIDE-BINDING, AND
RP MUTAGENESIS OF GLU-1195 AND TRP-1250.
RC STRAIN=Langeland / NCTC 10281 / Type F;
RX PubMed=19650874; DOI=10.1111/j.1471-4159.2009.06298.x;
RA Rummel A., Haefner K., Mahrhold S., Darashchonak N., Holt M., Jahn R.,
RA Beermann S., Karnath T., Bigalke H., Binz T.;
RT "Botulinum neurotoxins C, E and F bind gangliosides via a conserved binding
RT site prior to stimulation-dependent uptake with botulinum neurotoxin F
RT utilising the three isoforms of SV2 as second receptor.";
RL J. Neurochem. 110:1942-1954(2009).
RN [7]
RP FUNCTION (BOTULINUM NEUROTOXIN TYPE F), AND SV2 NOT RECEPTOR.
RC STRAIN=Type F;
RX PubMed=21483489; DOI=10.1371/journal.ppat.1002008;
RA Peng L., Tepp W.H., Johnson E.A., Dong M.;
RT "Botulinum neurotoxin D uses synaptic vesicle protein SV2 and gangliosides
RT as receptors.";
RL PLoS Pathog. 7:E1002008-E1002008(2011).
RN [8]
RP FUNCTION (BOTULINUM NEUROTOXIN F LIGHT CHAIN), SUBSTRATE SPECIFICITY, AND
RP BIOPHYSICOCHEMICAL PROPERTIES.
RC STRAIN=Langeland / NCTC 10281 / Type F;
RX PubMed=22289120; DOI=10.1111/j.1348-0421.2012.00434.x;
RA Yamamoto H., Ida T., Tsutsuki H., Mori M., Matsumoto T., Kohda T.,
RA Mukamoto M., Goshima N., Kozaki S., Ihara H.;
RT "Specificity of botulinum protease for human VAMP family proteins.";
RL Microbiol. Immunol. 56:245-253(2012).
RN [9]
RP REVIEW.
RX PubMed=28356439; DOI=10.1124/pr.116.012658;
RA Pirazzini M., Rossetto O., Eleopra R., Montecucco C.;
RT "Botulinum neurotoxins: Biology, pharmacology, and toxicology.";
RL Pharmacol. Rev. 69:200-235(2017).
RN [10] {ECO:0007744|PDB:3FUQ}
RP X-RAY CRYSTALLOGRAPHY (2.10 ANGSTROMS) OF 862-1278, FUNCTION (BOTULINUM
RP NEUROTOXIN TYPE F), FUNCTION (BOTULINUM NEUROTOXIN F HEAVY CHAIN),
RP INTERACTION WITH HOST SV2 AND SYT1, SUBCELLULAR LOCATION (BOTULINUM
RP NEUROTOXIN F LIGHT CHAIN), DOMAIN, GANGLIOSIDE-BINDING, AND MUTAGENESIS OF
RP GLU-1195; SER-1248; TRP-1250; TYR-1251 AND ASN-1254.
RC STRAIN=Langeland / NCTC 10281 / Type F;
RX PubMed=19476346; DOI=10.1021/bi9002138;
RA Fu Z., Chen C., Barbieri J.T., Kim J.J., Baldwin M.R.;
RT "Glycosylated SV2 and gangliosides as dual receptors for botulinum
RT neurotoxin serotype F.";
RL Biochemistry 48:5631-5641(2009).
RN [11] {ECO:0007744|PDB:3FIE, ECO:0007744|PDB:3FII}
RP X-RAY CRYSTALLOGRAPHY (2.10 ANGSTROMS) OF 1-419 IN COMPLEX WITH ZINC AND
RP VAMP2 INHIBITORS, FUNCTION (BOTULINUM NEUROTOXIN F LIGHT CHAIN), SUBSTRATE
RP SPECIFICITY, COFACTOR, AND MUTAGENESIS OF ARG-133; GLU-164; ARG-171 AND
RP TYR-316.
RC STRAIN=Langeland / NCTC 10281 / Type F;
RX PubMed=19543288; DOI=10.1038/nsmb.1626;
RA Agarwal R., Schmidt J.J., Stafford R.G., Swaminathan S.;
RT "Mode of VAMP substrate recognition and inhibition of Clostridium botulinum
RT neurotoxin F.";
RL Nat. Struct. Mol. Biol. 16:789-794(2009).
RN [12] {ECO:0007744|PDB:3RSJ}
RP X-RAY CRYSTALLOGRAPHY (2.00 ANGSTROMS) OF 866-1278 IN COMPLEX WITH
RP GANGLIOSIDE GD1A, FUNCTION (BOTULINUM NEUROTOXIN F HEAVY CHAIN),
RP GANGLIOSIDE-BINDING, AND MUTAGENESIS OF ARG-1111; HIS-1241 AND ARG-1256.
RC STRAIN=Langeland / NCTC 10281 / Type F;
RX PubMed=21849494; DOI=10.1074/jbc.m111.272054;
RA Benson M.A., Fu Z., Kim J.J., Baldwin M.R.;
RT "Unique ganglioside recognition strategies for clostridial neurotoxins.";
RL J. Biol. Chem. 286:34015-34022(2011).
CC -!- FUNCTION: [Botulinum neurotoxin type F]: Botulinum toxin causes flaccid
CC paralysis by inhibiting neurotransmitter (acetylcholine) release from
CC the presynaptic membranes of nerve terminals of the eukaryotic host
CC skeletal and autonomic nervous system, with frequent heart or
CC respiratory failure (PubMed:14423425). Precursor of botulinum
CC neurotoxin F which may have 2 coreceptors; complex polysialylated
CC gangliosides found on neural tissue and specific membrane-anchored
CC proteins found in synaptic vesicles. Receptor proteins are exposed on
CC host presynaptic cell membrane during neurotransmitter release, when
CC the toxin heavy chain (HC) binds to them (PubMed:19476346,
CC PubMed:19650874). Upon synaptic vesicle recycling the toxin is taken up
CC via the endocytic pathway. When the pH of the toxin-containing endosome
CC drops a structural rearrangement occurs so that the N-terminus of the
CC HC forms pores that allows the light chain (LC) to translocate into the
CC cytosol. Once in the cytosol the disulfide bond linking the 2 subunits
CC is reduced and LC cleaves its target protein on synaptic vesicles,
CC preventing their fusion with the cytoplasmic membrane and thus
CC neurotransmitter release (By similarity). Requires complex gangliosides
CC for full neurotoxicity (PubMed:19650874, PubMed:21483489). Electrical
CC stimulation increases uptake of toxin, presumably by transiently
CC exposing a receptor usually found in eukaryotic target synaptic
CC vesicles (PubMed:19476346, PubMed:19650874). Blocks neurotransitter
CC release by cleaving synaptobrevin-2/VAMP2 (PubMed:19476346). It is not
CC clear whether a synaptic vesicle protein acts as its receptor; there is
CC evidence for and against SV2 fulfilling this function (PubMed:19650874,
CC PubMed:21483489, PubMed:19476346). {ECO:0000250|UniProtKB:P0DPI0,
CC ECO:0000269|PubMed:14423425, ECO:0000269|PubMed:19476346,
CC ECO:0000269|PubMed:19650874, ECO:0000269|PubMed:21483489}.
CC -!- FUNCTION: [Botulinum neurotoxin F light chain]: Has protease activity
CC (PubMed:19476346, PubMed:19543288). After translocation into the
CC eukaryotic host cytosol, inhibits neurotransmitter release by acting as
CC a zinc endopeptidase that catalyzes the hydrolysis of the '58-Gln-|-
CC Lys-59' bond of synaptobrevin-2/VAMP2 and probably also the equivalent
CC 'Gln-|-Lys' sites in VAMP1 and VAMP3 (PubMed:19476346,
CC PubMed:19543288). Substrate specificity is conferred by multiple
CC interactions of LC with substrate (PubMed:19543288).
CC {ECO:0000250|UniProtKB:P30996, ECO:0000269|PubMed:19543288,
CC ECO:0000305|PubMed:19476346}.
CC -!- FUNCTION: [Botulinum neurotoxin F heavy chain]: Responsible for host
CC epithelial cell transcytosis, host nerve cell targeting and
CC translocation of light chain (LC) into host cytosol. Composed of 3
CC subdomains; the translocation domain (TD), and N-terminus and C-
CC terminus of the receptor-binding domain (RBD). The RBD is responsible
CC for the adherence of the toxin to the cell surface (PubMed:19476346,
CC PubMed:19650874). The N-terminus of the TD wraps an extended belt
CC around the perimeter of the LC, protecting Zn(2+) in the active site;
CC it may also prevent premature LC dissociation from the translocation
CC channel and protect toxin prior to translocation (By similarity).
CC Isolated HC binds to host synaptosomes and neurons, significantly
CC decreases uptake and toxicity of whole BoNT/F (PubMed:19476346,
CC PubMed:19650874). Interferes with uptake of BoNT/E and to a lesser
CC extent BoNT/C (PubMed:19650874). in vitro binds gangliosides GT1b, GD1b
CC and GD1a (PubMed:19650874, PubMed:19476346, PubMed:21849494). Binds to
CC synaptic vesicle glycoproteins SV2A, SV2B and SV2C which may serve as
CC coreceptors with gangliosides (PubMed:19650874, PubMed:19476346).
CC Interaction with SV2 proteins requires SV2 glycosylation
CC (PubMed:19476346). However knockout SV2A/SV2B mice still cleave
CC synaptobrevin, leaving the identification of its receptor unclear
CC (PubMed:21483489). {ECO:0000250|UniProtKB:P0DPI0,
CC ECO:0000269|PubMed:19476346, ECO:0000269|PubMed:19650874,
CC ECO:0000269|PubMed:21483489}.
CC -!- CATALYTIC ACTIVITY:
CC Reaction=Limited hydrolysis of proteins of the neuroexocytosis
CC apparatus, synaptobrevins, SNAP25 or syntaxin. No detected action on
CC small molecule substrates.; EC=3.4.24.69;
CC Evidence={ECO:0000305|PubMed:19476346};
CC -!- COFACTOR:
CC Name=Zn(2+); Xref=ChEBI:CHEBI:29105;
CC Evidence={ECO:0000269|PubMed:19543288};
CC Note=Binds 1 zinc ion per subunit (PubMed:19543288).
CC {ECO:0000269|PubMed:19543288};
CC -!- BIOPHYSICOCHEMICAL PROPERTIES:
CC Kinetic parameters:
CC KM=19.0 uM for over-expressed human VAMP1
CC {ECO:0000269|PubMed:22289120};
CC KM=24.5 uM for over-expressed human VAMP2
CC {ECO:0000269|PubMed:22289120};
CC KM=15.0 uM for over-expressed human VAMP3
CC {ECO:0000269|PubMed:22289120};
CC Note=kcat is 16.12, 34.37 and 28.57 sec(-1) for over-expressed human
CC VAMP1, VAMP2 and VAMP3 respectively. {ECO:0000269|PubMed:22289120};
CC -!- SUBUNIT: Heterodimer; disulfide-linked heterodimer of a light chain
CC (LC) and a heavy chain (HC). The LC has the proteolytic/pharmacological
CC activity, while the N- and C-terminal of the HC mediate channel
CC formation and toxin binding, respectively. Interacts with host synaptic
CC vesicle glycoproteins SV2A, SV2B and SV2C (PubMed:19650874). HC
CC interacts with a complex including at least host SV2 and synaptotagmin-
CC 1 (SYT1); copurification depends on glycosylation of SV2
CC (PubMed:19476346). {ECO:0000269|PubMed:19476346,
CC ECO:0000269|PubMed:19650874}.
CC -!- INTERACTION:
CC A7GBG3; P63027: VAMP2; Xeno; NbExp=2; IntAct=EBI-7604673, EBI-520113;
CC -!- SUBCELLULAR LOCATION: [Botulinum neurotoxin type F]: Secreted
CC {ECO:0000250|UniProtKB:P0DPI0}.
CC -!- SUBCELLULAR LOCATION: [Botulinum neurotoxin F light chain]: Secreted
CC {ECO:0000250|UniProtKB:P0DPI0}. Host cytoplasm, host cytosol
CC {ECO:0000305|PubMed:19476346, ECO:0000305|PubMed:19543288}.
CC -!- SUBCELLULAR LOCATION: [Botulinum neurotoxin F heavy chain]: Secreted
CC {ECO:0000250|UniProtKB:P0DPI0}. Host synapse, host presynaptic cell
CC membrane {ECO:0000250|UniProtKB:P0DPI0}. Host cytoplasmic vesicle, host
CC secretory vesicle, host synaptic vesicle membrane
CC {ECO:0000305|PubMed:19476346}; Multi-pass membrane protein
CC {ECO:0000305}.
CC -!- DOMAIN: [Botulinum neurotoxin F light chain]: Has protease activity
CC (PubMed:19476346, PubMed:19543288). {ECO:0000269|PubMed:19543288,
CC ECO:0000305|PubMed:19476346}.
CC -!- DOMAIN: [Botulinum neurotoxin F heavy chain]: Has 3 functional domains;
CC the translocation domain (TD) and the receptor-binding domain (RBD)
CC which is further subdivided into N- and C-terminal domains (N-RBD and
CC C-RBD) (PubMed:19476346). The N-terminus of the TD wraps an extended
CC belt around the perimeter of the LC, protecting Zn(2+) in the active
CC site and may be a pseudosubstrate inhibitor which serves as an
CC intramolecular chaperone for the LC prior to its translocation into the
CC host cytosol (By similarity). The RBD binds transiently exposed
CC coreceptors on the host presynaptic cell membrane (PubMed:19476346,
CC PubMed:19650874). {ECO:0000250|UniProtKB:P0DPI0,
CC ECO:0000269|PubMed:19650874, ECO:0000305|PubMed:19476346}.
CC -!- MISCELLANEOUS: There are seven antigenically distinct forms of
CC botulinum neurotoxin: Types A, B, C, D, E, F, and G; new subtypes are
CC quite frequent.
CC -!- MISCELLANEOUS: Botulism poisoning is usually food-borne, either by
CC ingesting toxin or bacterial-contaminated food, or less frequently by
CC inhalation poisoning. In both cases the neurotoxin binds to the apical
CC surface of epithelial cells in the gut or airway. Toxin undergoes
CC receptor-mediated endocytosis (using a different receptor than on
CC target nerve cells), transcytosis across the epithelial cells and
CC release into the general circulation. Once in the general circulation
CC it binds to its target cells. {ECO:0000250|UniProtKB:P0DPI0}.
CC -!- MISCELLANEOUS: Strain Langeland / NCTC 10281 / Type F was isolated from
CC home-made liver paste associated with human botulism in Denmark in 1958
CC (PubMed:14423425). It is type F1 (PubMed:20511432).
CC {ECO:0000269|PubMed:14423425, ECO:0000269|PubMed:20511432}.
CC -!- SIMILARITY: Belongs to the peptidase M27 family. {ECO:0000305}.
CC -!- CAUTION: It is not clear whether a synaptic vesicle protein acts as its
CC receptor; there is evidence for and against SV2 fulfilling this
CC function (PubMed:19650874, PubMed:21483489, PubMed:19476346).
CC {ECO:0000269|PubMed:19476346, ECO:0000269|PubMed:19650874,
CC ECO:0000269|PubMed:21483489}.
CC -!- WEB RESOURCE: Name=BotDB - A Database Resource for Clostridial
CC Neurotoxins;
CC URL="https://botdb.abcc.ncifcrf.gov/";
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DR EMBL; X81714; CAA57358.1; -; Genomic_DNA.
DR EMBL; GU213203; ADA79551.1; -; Genomic_DNA.
DR EMBL; X70821; CAA50152.1; -; Genomic_DNA.
DR EMBL; CP000728; ABS41202.1; -; Genomic_DNA.
DR PIR; S48110; S48110.
DR RefSeq; WP_011987710.1; NC_009699.1.
DR PDB; 3FIE; X-ray; 2.10 A; A/B=1-419.
DR PDB; 3FII; X-ray; 2.17 A; A=1-419.
DR PDB; 3FUQ; X-ray; 2.10 A; A=862-1278.
DR PDB; 3RSJ; X-ray; 2.00 A; A/B/C/D=866-1278.
DR PDBsum; 3FIE; -.
DR PDBsum; 3FII; -.
DR PDBsum; 3FUQ; -.
DR PDBsum; 3RSJ; -.
DR AlphaFoldDB; A7GBG3; -.
DR SMR; A7GBG3; -.
DR IntAct; A7GBG3; 1.
DR MINT; A7GBG3; -.
DR UniLectin; A7GBG3; -.
DR ABCD; A7GBG3; 10 sequenced antibodies.
DR EnsemblBacteria; ABS41202; ABS41202; CLI_0851.
DR KEGG; cbf:CLI_0851; -.
DR HOGENOM; CLU_262205_0_0_9; -.
DR OMA; DEGYNKA; -.
DR BRENDA; 3.4.24.69; 1462.
DR Proteomes; UP000002410; Chromosome.
DR GO; GO:0005576; C:extracellular region; IEA:UniProtKB-SubCell.
DR GO; GO:0044161; C:host cell cytoplasmic vesicle; IEA:UniProtKB-SubCell.
DR GO; GO:0044164; C:host cell cytosol; IEA:UniProtKB-SubCell.
DR GO; GO:0044156; C:host cell junction; IEA:UniProtKB-KW.
DR GO; GO:0044231; C:host cell presynaptic membrane; IEA:UniProtKB-SubCell.
DR GO; GO:0016020; C:membrane; IEA:UniProtKB-KW.
DR GO; GO:0008289; F:lipid binding; IEA:UniProtKB-KW.
DR GO; GO:0004222; F:metalloendopeptidase activity; IEA:UniProtKB-EC.
DR GO; GO:0008320; F:protein transmembrane transporter activity; IEA:InterPro.
DR GO; GO:0090729; F:toxin activity; IEA:UniProtKB-KW.
DR GO; GO:0008270; F:zinc ion binding; IEA:InterPro.
DR GO; GO:0046929; P:negative regulation of neurotransmitter secretion; IEA:InterPro.
DR GO; GO:0006508; P:proteolysis; IEA:UniProtKB-KW.
DR Gene3D; 1.20.1120.10; -; 1.
DR InterPro; IPR000395; Bot/tetX_LC.
DR InterPro; IPR036248; Clostridium_toxin_transloc.
DR InterPro; IPR013320; ConA-like_dom_sf.
DR InterPro; IPR011065; Kunitz_inhibitor_STI-like_sf.
DR InterPro; IPR013104; Toxin_rcpt-bd_C.
DR InterPro; IPR012928; Toxin_rcpt-bd_N.
DR InterPro; IPR012500; Toxin_trans.
DR Pfam; PF01742; Peptidase_M27; 1.
DR Pfam; PF07951; Toxin_R_bind_C; 1.
DR Pfam; PF07953; Toxin_R_bind_N; 1.
DR Pfam; PF07952; Toxin_trans; 1.
DR PRINTS; PR00760; BONTOXILYSIN.
DR SUPFAM; SSF49899; SSF49899; 1.
DR SUPFAM; SSF50386; SSF50386; 1.
DR SUPFAM; SSF58091; SSF58091; 1.
DR PROSITE; PS00142; ZINC_PROTEASE; 1.
PE 1: Evidence at protein level;
KW 3D-structure; Coiled coil; Disulfide bond; Host cell membrane;
KW Host cytoplasm; Host cytoplasmic vesicle; Host membrane; Host synapse;
KW Hydrolase; Lipid-binding; Membrane; Metal-binding; Metalloprotease;
KW Neurotoxin; Protease; Secreted; Toxin; Virulence; Zinc.
FT CHAIN 1..1278
FT /note="Botulinum neurotoxin type F"
FT /id="PRO_0000444908"
FT CHAIN 1..436
FT /note="Botulinum neurotoxin F light chain"
FT /id="PRO_0000444909"
FT CHAIN 437..1278
FT /note="Botulinum neurotoxin F heavy chain"
FT /id="PRO_0000444910"
FT REGION 440..862
FT /note="Translocation domain (TD)"
FT /evidence="ECO:0000250|UniProtKB:P0DPI0"
FT REGION 485..534
FT /note="Belt"
FT /evidence="ECO:0000250|UniProtKB:P0DPI0"
FT REGION 864..1085
FT /note="N-terminus of receptor binding domain (N-RBD)"
FT /evidence="ECO:0000305|PubMed:19476346"
FT REGION 1086..1278
FT /note="C-terminus of receptor binding domain (C-RBD)"
FT /evidence="ECO:0000305|PubMed:19476346"
FT REGION 1240..1241
FT /note="Host ganglioside GD1a binding"
FT /evidence="ECO:0000269|PubMed:21849494,
FT ECO:0007744|PDB:3RSJ"
FT REGION 1248..1250
FT /note="Host ganglioside GD1a binding"
FT /evidence="ECO:0000269|PubMed:21849494,
FT ECO:0007744|PDB:3RSJ"
FT COILED 717..783
FT /evidence="ECO:0000255"
FT MOTIF 1245..1248
FT /note="Host ganglioside-binding motif"
FT /evidence="ECO:0000250|UniProtKB:P0DPI0,
FT ECO:0000305|PubMed:19476346, ECO:0000305|PubMed:19650874"
FT ACT_SITE 228
FT /evidence="ECO:0000255|PROSITE-ProRule:PRU10095"
FT BINDING 227
FT /ligand="Zn(2+)"
FT /ligand_id="ChEBI:CHEBI:29105"
FT /ligand_note="catalytic"
FT /evidence="ECO:0000255|PROSITE-ProRule:PRU10095,
FT ECO:0000269|PubMed:19543288, ECO:0007744|PDB:3FIE,
FT ECO:0007744|PDB:3FII"
FT BINDING 231
FT /ligand="Zn(2+)"
FT /ligand_id="ChEBI:CHEBI:29105"
FT /ligand_note="catalytic"
FT /evidence="ECO:0000255|PROSITE-ProRule:PRU10095,
FT ECO:0000269|PubMed:19543288, ECO:0007744|PDB:3FIE,
FT ECO:0007744|PDB:3FII"
FT BINDING 266
FT /ligand="Zn(2+)"
FT /ligand_id="ChEBI:CHEBI:29105"
FT /ligand_note="catalytic"
FT /evidence="ECO:0000269|PubMed:19543288,
FT ECO:0007744|PDB:3FIE, ECO:0007744|PDB:3FII"
FT BINDING 1111
FT /ligand="ganglioside GD1a (d18:1(4E))"
FT /ligand_id="ChEBI:CHEBI:78445"
FT /ligand_note="host ganglioside"
FT /evidence="ECO:0000269|PubMed:21849494"
FT BINDING 1195
FT /ligand="ganglioside GD1a (d18:1(4E))"
FT /ligand_id="ChEBI:CHEBI:78445"
FT /ligand_note="host ganglioside"
FT /evidence="ECO:0000269|PubMed:21849494,
FT ECO:0007744|PDB:3RSJ"
FT BINDING 1256
FT /ligand="ganglioside GD1a (d18:1(4E))"
FT /ligand_id="ChEBI:CHEBI:78445"
FT /ligand_note="host ganglioside"
FT /evidence="ECO:0000269|PubMed:21849494,
FT ECO:0007744|PDB:3RSJ"
FT DISULFID 429..445
FT /note="Interchain (between light and heavy chains)"
FT /evidence="ECO:0000250|UniProtKB:P0DPI0, ECO:0000305"
FT MUTAGEN 133
FT /note="R->A: Dramatically decreased cleavage of host
FT synaptobrevin-2 (VAMP2)."
FT /evidence="ECO:0000269|PubMed:19543288"
FT MUTAGEN 133
FT /note="R->K: Dramatically decreased cleavage of VAMP2."
FT /evidence="ECO:0000269|PubMed:19543288"
FT MUTAGEN 164
FT /note="E->A: Significantly decreased cleavage of VAMP2."
FT /evidence="ECO:0000269|PubMed:19543288"
FT MUTAGEN 171
FT /note="R->K: Dramatically decreased cleavage of VAMP2."
FT /evidence="ECO:0000269|PubMed:19543288"
FT MUTAGEN 316
FT /note="Y->A: Slightly decreased cleavage of VAMP2."
FT /evidence="ECO:0000269|PubMed:19543288"
FT MUTAGEN 1111
FT /note="R->A: Decreased heavy chain (HC) binding to
FT ganglioside GD1a. Dramatically decreased HC binding to
FT GD1a; when associated with A-1256. Significantly decreased;
FT when associated with K-1241 and A-1256, which partially
FT restores binding."
FT /evidence="ECO:0000269|PubMed:21849494"
FT MUTAGEN 1195
FT /note="E->A: Significantly decreased binding of heavy chain
FT to synaptosomes, significantly decreased binding to
FT ganglioside GT1b."
FT /evidence="ECO:0000269|PubMed:19476346,
FT ECO:0000269|PubMed:19650874"
FT MUTAGEN 1241
FT /note="H->K: Dramatically decreased heavy chain (HC)
FT binding to ganglioside GD1a. Significantly decreased HC
FT binding to GD1a; when associated with A-1111 and A-1256,
FT partially restores binding."
FT /evidence="ECO:0000269|PubMed:21849494"
FT MUTAGEN 1248
FT /note="S->A: Significantly decreased heavy chain binding to
FT ganglioside GT1b."
FT /evidence="ECO:0000269|PubMed:19476346"
FT MUTAGEN 1250
FT /note="W->L: Significantly decreased binding of heavy chain
FT (HC) to synaptosomes, significantly decreased binding to
FT ganglioside GT1b. HC no longer inhibits BoNT/F whole-toxin
FT uptake and toxicity. Loss of HC binding to synaptic
FT vesicles, greatly decreased binding to neurons."
FT /evidence="ECO:0000269|PubMed:19476346,
FT ECO:0000269|PubMed:19650874"
FT MUTAGEN 1251
FT /note="Y->F: Decreased heavy chain binding to ganglioside
FT GT1b."
FT /evidence="ECO:0000269|PubMed:19476346"
FT MUTAGEN 1254
FT /note="N->A: Wild-type heavy chain binding to ganglioside
FT GT1b."
FT /evidence="ECO:0000269|PubMed:19476346"
FT MUTAGEN 1256
FT /note="R->A: Decreased heavy chain (HC) binding to
FT ganglioside GD1a. Dramatically decreased HC binding to
FT GD1a; when associated with A-1111. Significantly decreased;
FT when associated with A-1111 and K-1241, which partially
FT restores binding."
FT /evidence="ECO:0000269|PubMed:21849494"
FT STRAND 16..22
FT /evidence="ECO:0007829|PDB:3FIE"
FT HELIX 28..30
FT /evidence="ECO:0007829|PDB:3FII"
FT STRAND 34..40
FT /evidence="ECO:0007829|PDB:3FIE"
FT STRAND 43..49
FT /evidence="ECO:0007829|PDB:3FIE"
FT HELIX 56..59
FT /evidence="ECO:0007829|PDB:3FIE"
FT TURN 67..69
FT /evidence="ECO:0007829|PDB:3FII"
FT TURN 75..78
FT /evidence="ECO:0007829|PDB:3FIE"
FT HELIX 81..99
FT /evidence="ECO:0007829|PDB:3FIE"
FT HELIX 102..113
FT /evidence="ECO:0007829|PDB:3FIE"
FT TURN 133..135
FT /evidence="ECO:0007829|PDB:3FIE"
FT STRAND 136..140
FT /evidence="ECO:0007829|PDB:3FIE"
FT STRAND 146..150
FT /evidence="ECO:0007829|PDB:3FIE"
FT STRAND 152..157
FT /evidence="ECO:0007829|PDB:3FIE"
FT STRAND 166..169
FT /evidence="ECO:0007829|PDB:3FIE"
FT HELIX 182..184
FT /evidence="ECO:0007829|PDB:3FIE"
FT STRAND 191..194
FT /evidence="ECO:0007829|PDB:3FIE"
FT STRAND 199..204
FT /evidence="ECO:0007829|PDB:3FIE"
FT STRAND 216..218
FT /evidence="ECO:0007829|PDB:3FIE"
FT HELIX 221..236
FT /evidence="ECO:0007829|PDB:3FIE"
FT HELIX 241..245
FT /evidence="ECO:0007829|PDB:3FIE"
FT STRAND 247..251
FT /evidence="ECO:0007829|PDB:3FIE"
FT TURN 253..255
FT /evidence="ECO:0007829|PDB:3FIE"
FT STRAND 258..263
FT /evidence="ECO:0007829|PDB:3FIE"
FT HELIX 264..270
FT /evidence="ECO:0007829|PDB:3FIE"
FT HELIX 272..277
FT /evidence="ECO:0007829|PDB:3FIE"
FT HELIX 280..302
FT /evidence="ECO:0007829|PDB:3FIE"
FT HELIX 313..323
FT /evidence="ECO:0007829|PDB:3FIE"
FT STRAND 326..328
FT /evidence="ECO:0007829|PDB:3FIE"
FT STRAND 334..336
FT /evidence="ECO:0007829|PDB:3FIE"
FT HELIX 338..349
FT /evidence="ECO:0007829|PDB:3FIE"
FT HELIX 353..359
FT /evidence="ECO:0007829|PDB:3FIE"
FT STRAND 366..368
FT /evidence="ECO:0007829|PDB:3FIE"
FT STRAND 374..376
FT /evidence="ECO:0007829|PDB:3FIE"
FT TURN 383..385
FT /evidence="ECO:0007829|PDB:3FIE"
FT TURN 388..390
FT /evidence="ECO:0007829|PDB:3FIE"
FT HELIX 395..404
FT /evidence="ECO:0007829|PDB:3FIE"
FT TURN 406..408
FT /evidence="ECO:0007829|PDB:3FIE"
FT HELIX 410..415
FT /evidence="ECO:0007829|PDB:3FIE"
FT HELIX 867..869
FT /evidence="ECO:0007829|PDB:3RSJ"
FT STRAND 870..876
FT /evidence="ECO:0007829|PDB:3RSJ"
FT STRAND 878..883
FT /evidence="ECO:0007829|PDB:3RSJ"
FT STRAND 885..887
FT /evidence="ECO:0007829|PDB:3RSJ"
FT STRAND 890..895
FT /evidence="ECO:0007829|PDB:3RSJ"
FT STRAND 906..920
FT /evidence="ECO:0007829|PDB:3RSJ"
FT TURN 923..925
FT /evidence="ECO:0007829|PDB:3RSJ"
FT STRAND 934..941
FT /evidence="ECO:0007829|PDB:3RSJ"
FT HELIX 948..950
FT /evidence="ECO:0007829|PDB:3RSJ"
FT STRAND 955..962
FT /evidence="ECO:0007829|PDB:3RSJ"
FT TURN 963..965
FT /evidence="ECO:0007829|PDB:3RSJ"
FT STRAND 966..973
FT /evidence="ECO:0007829|PDB:3RSJ"
FT STRAND 976..982
FT /evidence="ECO:0007829|PDB:3RSJ"
FT STRAND 988..994
FT /evidence="ECO:0007829|PDB:3RSJ"
FT STRAND 997..1001
FT /evidence="ECO:0007829|PDB:3RSJ"
FT STRAND 1007..1015
FT /evidence="ECO:0007829|PDB:3RSJ"
FT STRAND 1019..1025
FT /evidence="ECO:0007829|PDB:3RSJ"
FT STRAND 1028..1034
FT /evidence="ECO:0007829|PDB:3RSJ"
FT STRAND 1045..1053
FT /evidence="ECO:0007829|PDB:3RSJ"
FT STRAND 1059..1070
FT /evidence="ECO:0007829|PDB:3RSJ"
FT HELIX 1074..1083
FT /evidence="ECO:0007829|PDB:3RSJ"
FT STRAND 1095..1097
FT /evidence="ECO:0007829|PDB:3RSJ"
FT STRAND 1099..1101
FT /evidence="ECO:0007829|PDB:3RSJ"
FT STRAND 1103..1111
FT /evidence="ECO:0007829|PDB:3RSJ"
FT STRAND 1114..1118
FT /evidence="ECO:0007829|PDB:3RSJ"
FT STRAND 1120..1128
FT /evidence="ECO:0007829|PDB:3RSJ"
FT STRAND 1131..1134
FT /evidence="ECO:0007829|PDB:3RSJ"
FT TURN 1135..1137
FT /evidence="ECO:0007829|PDB:3RSJ"
FT STRAND 1138..1141
FT /evidence="ECO:0007829|PDB:3RSJ"
FT STRAND 1148..1153
FT /evidence="ECO:0007829|PDB:3RSJ"
FT STRAND 1171..1178
FT /evidence="ECO:0007829|PDB:3RSJ"
FT STRAND 1181..1187
FT /evidence="ECO:0007829|PDB:3RSJ"
FT STRAND 1193..1202
FT /evidence="ECO:0007829|PDB:3RSJ"
FT STRAND 1212..1219
FT /evidence="ECO:0007829|PDB:3RSJ"
FT STRAND 1222..1229
FT /evidence="ECO:0007829|PDB:3RSJ"
FT STRAND 1234..1248
FT /evidence="ECO:0007829|PDB:3RSJ"
FT HELIX 1249..1253
FT /evidence="ECO:0007829|PDB:3RSJ"
FT STRAND 1266..1269
FT /evidence="ECO:0007829|PDB:3RSJ"
SQ SEQUENCE 1278 AA; 147075 MW; A1BE1318431D6918 CRC64;
MPVVINSFNY NDPVNDDTIL YMQIPYEEKS KKYYKAFEIM RNVWIIPERN TIGTDPSDFD
PPASLENGSS AYYDPNYLTT DAEKDRYLKT TIKLFKRINS NPAGEVLLQE ISYAKPYLGN
EHTPINEFHP VTRTTSVNIK SSTNVKSSII LNLLVLGAGP DIFENSSYPV RKLMDSGGVY
DPSNDGFGSI NIVTFSPEYE YTFNDISGGY NSSTESFIAD PAISLAHELI HALHGLYGAR
GVTYKETIKV KQAPLMIAEK PIRLEEFLTF GGQDLNIITS AMKEKIYNNL LANYEKIATR
LSRVNSAPPE YDINEYKDYF QWKYGLDKNA DGSYTVNENK FNEIYKKLYS FTEIDLANKF
KVKCRNTYFI KYGFLKVPNL LDDDIYTVSE GFNIGNLAVN NRGQNIKLNP KIIDSIPDKG
LVEKIVKFCK SVIPRKGTKA PPRLCIRVNN RELFFVASES SYNENDINTP KEIDDTTNLN
NNYRNNLDEV ILDYNSETIP QISNQTLNTL VQDDSYVPRY DSNGTSEIEE HNVVDLNVFF
YLHAQKVPEG ETNISLTSSI DTALSEESQV YTFFSSEFIN TINKPVHAAL FISWINQVIR
DFTTEATQKS TFDKIADISL VVPYVGLALN IGNEVQKENF KEAFELLGAG ILLEFVPELL
IPTILVFTIK SFIGSSENKN KIIKAINNSL MERETKWKEI YSWIVSNWLT RINTQFNKRK
EQMYQALQNQ VDAIKTVIEY KYNNYTSDER NRLESEYNIN NIREELNKKV SLAMENIERF
ITESSIFYLM KLINEAKVSK LREYDEGVKE YLLDYISEHR SILGNSVQEL NDLVTSTLNN
SIPFELSSYT NDKILILYFN KLYKKIKDNS ILDMRYENNK FIDISGYGSN ISINGDVYIY
STNRNQFGIY SSKPSEVNIA QNNDIIYNGR YQNFSISFWV RIPKYFNKVN LNNEYTIIDC
IRNNNSGWKI SLNYNKIIWT LQDTAGNNQK LVFNYTQMIS ISDYINKWIF VTITNNRLGN
SRIYINGNLI DEKSISNLGD IHVSDNILFK IVGCNDTRYV GIRYFKVFDT ELGKTEIETL
YSDEPDPSIL KDFWGNYLLY NKRYYLLNLL RTDKSITQNS NFLNINQQRG VYQKPNIFSN
TRLYTGVEVI IRKNGSTDIS NTDNFVRKND LAYINVVDRD VEYRLYADIS IAKPEKIIKL
IRTSNSNNSL GQIIVMDSIG NNCTMNFQNN NGGNIGLLGF HSNNLVASSW YYNNIRKNTS
SNGCFWSFIS KEHGWQEN
