BXA1_CLOBO
ID BXA1_CLOBO Reviewed; 1296 AA.
AC P0DPI0; A5HZZ9; A7G1U9; P01561; P10845; P18639;
DT 18-JUL-2018, integrated into UniProtKB/Swiss-Prot.
DT 18-JUL-2018, sequence version 1.
DT 03-AUG-2022, entry version 25.
DE RecName: Full=Botulinum neurotoxin type A;
DE Short=BoNT/A;
DE AltName: Full=Bontoxilysin-A;
DE Short=BOTOX;
DE AltName: Full=Botulinum neurotoxin type A1;
DE Contains:
DE RecName: Full=Botulinum neurotoxin A light chain;
DE Short=LC;
DE EC=3.4.24.69 {ECO:0000269|PubMed:8243676};
DE Contains:
DE RecName: Full=Botulinum neurotoxin A heavy chain;
DE Short=HC;
DE Flags: Precursor;
GN Name=botA {ECO:0000303|PubMed:2185020};
GN Synonyms=atx {ECO:0000303|PubMed:8521962},
GN bonT {ECO:0000303|PubMed:8863443};
OS Clostridium botulinum.
OC Bacteria; Firmicutes; Clostridia; Eubacteriales; Clostridiaceae;
OC Clostridium.
OX NCBI_TaxID=1491;
RN [1]
RP NUCLEOTIDE SEQUENCE [GENOMIC DNA].
RC STRAIN=NCTC 2916 / Type A;
RX PubMed=2185020; DOI=10.1111/j.1432-1033.1990.tb15461.x;
RA Thompson D.E., Brehm J.K., Oultram J.D., Swinfield T.-J., Shone C.C.,
RA Atkinson T., Melling J., Minton N.P.;
RT "The complete amino acid sequence of the Clostridium botulinum type A
RT neurotoxin, deduced by nucleotide sequence analysis of the encoding gene.";
RL Eur. J. Biochem. 189:73-81(1990).
RN [2]
RP NUCLEOTIDE SEQUENCE [GENOMIC DNA].
RC STRAIN=62A / Type A;
RX PubMed=2160960; DOI=10.1016/s0021-9258(19)38824-6;
RA Binz T., Kurazono H., Wille M., Frevert J., Wernars K., Niemann H.;
RT "The complete sequence of botulinum neurotoxin type A and comparison with
RT other clostridial neurotoxins.";
RL J. Biol. Chem. 265:9153-9158(1990).
RN [3]
RP NUCLEOTIDE SEQUENCE [GENOMIC DNA] OF 1-66.
RC STRAIN=62A / Type A;
RX PubMed=8863443; DOI=10.1099/00207713-46-4-1105;
RA East A.K., Bhandari M., Stacey J.M., Campbell K.D., Collins M.D.;
RT "Organization and phylogenetic interrelationships of genes encoding
RT components of the botulinum toxin complex in proteolytic Clostridium
RT botulinum types A, B, and F: evidence of chimeric sequences in the gene
RT encoding the nontoxic nonhemagglutinin component.";
RL Int. J. Syst. Bacteriol. 46:1105-1112(1996).
RN [4]
RP NUCLEOTIDE SEQUENCE [GENOMIC DNA] OF 1-19.
RC STRAIN=NIH / Type A;
RX PubMed=8521962; DOI=10.1016/0014-5793(95)01241-5;
RA Fujita R., Fujinaga Y., Inoue K., Nakajima H., Kumon H., Oguma K.;
RT "Molecular characterization of two forms of nontoxic-nonhemagglutinin
RT components of Clostridium botulinum type A progenitor toxins.";
RL FEBS Lett. 376:41-44(1995).
RN [5]
RP PROTEIN SEQUENCE OF 2-18 AND 449-458, AND SUBCELLULAR LOCATION (BOTULINUM
RP NEUROTOXIN A LIGHT CHAIN AND BOTULINUM NEUROTOXIN A HEAVY CHAIN).
RC STRAIN=Type A;
RX PubMed=6370252; DOI=10.1016/0006-291x(84)90858-1;
RA Schmidt J.J., Sartymoorthy V., Dasgupta B.R.;
RT "Partial amino acid sequence of the heavy and light chains of botulinum
RT neurotoxin type A.";
RL Biochem. Biophys. Res. Commun. 119:900-904(1984).
RN [6]
RP PROTEIN SEQUENCE OF 2-47.
RA Dasgupta B.R., Foley J., Niece R.;
RT "Partial sequence of the light chain of botulinum neurotoxin type A.";
RL Biochemistry 26:4162-4162(1987).
RN [7]
RP PROTEIN SEQUENCE OF 2-6 AND 445-457, AND PROCESSING.
RC STRAIN=Type A;
RX PubMed=2126206; DOI=10.1016/0300-9084(90)90048-l;
RA Dasgupta B.R., Dekleva M.L.;
RT "Botulinum neurotoxin type A: sequence of amino acids at the N-terminus and
RT around the nicking site.";
RL Biochimie 72:661-664(1990).
RN [8]
RP PROTEIN SEQUENCE OF 251-257; 267-273 AND 420-448, COFACTOR, AND PROTEOLYTIC
RP CLEAVAGE.
RX PubMed=11565902; DOI=10.1023/a:1010952025677;
RA Ahmed S.A., Byrne M.P., Jensen M., Hines H.B., Brueggemann E., Smith L.A.;
RT "Enzymatic autocatalysis of botulinum A neurotoxin light chain.";
RL J. Protein Chem. 20:221-231(2001).
RN [9]
RP PROTEIN SEQUENCE OF 449-483.
RC STRAIN=NCTC 2916 / Type A;
RX PubMed=3896784; DOI=10.1111/j.1432-1033.1985.tb09070.x;
RA Shone C.C., Hambleton P., Melling J.;
RT "Inactivation of Clostridium botulinum type A neurotoxin by trypsin and
RT purification of two tryptic fragments. Proteolytic action near the COOH-
RT terminus of the heavy subunit destroys toxin-binding activity.";
RL Eur. J. Biochem. 151:75-82(1985).
RN [10]
RP FUNCTION (BOTULINUM NEUROTOXIN TYPE A).
RC STRAIN=4587 / Type A;
RX PubMed=15394302; DOI=10.1113/jphysiol.1949.sp004364;
RA Burgen A.S., Dickens F., Zatman L.J.;
RT "The action of botulinum toxin on the neuro-muscular junction.";
RL J. Physiol. (Lond.) 109:10-24(1949).
RN [11]
RP FUNCTION (BOTULINUM NEUROTOXIN A HEAVY CHAIN), SUBCELLULAR LOCATION
RP (BOTULINUM NEUROTOXIN TYPE A AND BOTULINUM NEUROTOXIN A HEAVY CHAIN), AND
RP ACTIVITY REGULATION.
RC STRAIN=Type A;
RX PubMed=6694738; DOI=10.1038/307457a0;
RA Dolly J.O., Black J., Williams R.S., Melling J.;
RT "Acceptors for botulinum neurotoxin reside on motor nerve terminals and
RT mediate its internalization.";
RL Nature 307:457-460(1984).
RN [12]
RP FUNCTION (BOTULINUM NEUROTOXIN A HEAVY CHAIN), AND DOMAIN.
RC STRAIN=Type A;
RX PubMed=2446925; DOI=10.1016/0014-5793(87)80562-8;
RA Blaustein R.O., Germann W.J., Finkelstein A., DasGupta B.R.;
RT "The N-terminal half of the heavy chain of botulinum type A neurotoxin
RT forms channels in planar phospholipid bilayers.";
RL FEBS Lett. 226:115-120(1987).
RN [13]
RP IDENTIFICATION AS A ZINC-BINDING PROTEIN, AND COFACTOR.
RC STRAIN=Type A;
RX PubMed=1429690; DOI=10.1016/s0021-9258(18)35863-0;
RA Schiavo G., Rossetto O., Santucci A., Dasgupta B.R., Montecucco C.;
RT "Botulinum neurotoxins are zinc proteins.";
RL J. Biol. Chem. 267:23479-23483(1992).
RN [14]
RP HOST RANGE, AND EPIDEMIOLOGY.
RX PubMed=1431246; DOI=10.1093/infdis/166.6.1281;
RA Woodruff B.A., Griffin P.M., McCroskey L.M., Smart J.F., Wainwright R.B.,
RA Bryant R.G., Hutwagner L.C., Hatheway C.L.;
RT "Clinical and laboratory comparison of botulism from toxin types A, B, and
RT E in the United States, 1975-1988.";
RL J. Infect. Dis. 166:1281-1286(1992).
RN [15]
RP FUNCTION (BOTULINUM NEUROTOXIN A LIGHT CHAIN), IDENTIFICATION OF SUBSTRATE,
RP CATALYTIC ACTIVITY, AND SUBCELLULAR LOCATION (BOTULINUM NEUROTOXIN A LIGHT
RP CHAIN).
RC STRAIN=Type A;
RX PubMed=8243676; DOI=10.1016/0014-5793(93)80448-4;
RA Schiavo G., Santtuci A., Dasgupta B.R., Mehta P.P., Jontes J.,
RA Benfenati F., Wilson M.C., Montecucco C.;
RT "Botulinum neurotoxins serotypes A and E cleave SNAP-25 at distinct COOH-
RT terminal peptide bonds.";
RL FEBS Lett. 335:99-103(1993).
RN [16]
RP FUNCTION (BOTULINUM NEUROTOXIN TYPE A AND BOTULINUM NEUROTOXIN A LIGHT
RP CHAIN), CATALYTIC ACTIVITY, COFACTOR, SUBUNIT, SUBCELLULAR LOCATION
RP (BOTULINUM NEUROTOXIN A LIGHT CHAIN), DOMAIN, AND MUTAGENESIS OF HIS-227.
RC STRAIN=63A / Type A;
RX PubMed=7578132; DOI=10.1021/bi00046a025;
RA Zhou L., de Paiva A., Liu D., Aoki R., Dolly J.O.;
RT "Expression and purification of the light chain of botulinum neurotoxin A:
RT a single mutation abolishes its cleavage of SNAP-25 and neurotoxicity after
RT reconstitution with the heavy chain.";
RL Biochemistry 34:15175-15181(1995).
RN [17]
RP SUBCELLULAR LOCATION (BOTULINUM NEUROTOXIN TYPE A), AND INDUCTION.
RC STRAIN=62A / Type A;
RX PubMed=7592120; DOI=10.1111/j.1365-2672.1995.tb03135.x;
RA Call J.E., Cooke P.H., Miller A.J.;
RT "In situ characterization of Clostridium botulinum neurotoxin synthesis and
RT export.";
RL J. Appl. Bacteriol. 79:257-263(1995).
RN [18]
RP FUNCTION (BOTULINUM NEUROTOXIN A HEAVY CHAIN), AND SUBCELLULAR LOCATION
RP (BOTULINUM NEUROTOXIN A HEAVY CHAIN).
RC STRAIN=62A / P64 / Type A;
RX PubMed=10413679; DOI=10.1242/jcs.112.16.2715;
RA Lalli G., Herreros J., Osborne S.L., Montecucco C., Rossetto O.,
RA Schiavo G.;
RT "Functional characterisation of tetanus and botulinum neurotoxins binding
RT domains.";
RL J. Cell Sci. 112:2715-2724(1999).
RN [19]
RP FUNCTION (BOTULINUM NEUROTOXIN A LIGHT CHAIN), SUBSTRATE SPECIFICITY,
RP CATALYTIC ACTIVITY, AND SUBCELLULAR LOCATION (BOTULINUM NEUROTOXIN A LIGHT
RP CHAIN).
RC STRAIN=62A / P64 / Type A;
RX PubMed=9886085; DOI=10.1046/j.1471-4159.1999.0720327.x;
RA Vaidyanathan V.V., Yoshino K., Jahnz M., Doerries C., Bade S.,
RA Nauenburg S., Niemann H., Binz T.;
RT "Proteolysis of SNAP-25 isoforms by botulinum neurotoxin types A, C, and E:
RT domains and amino acid residues controlling the formation of enzyme-
RT substrate complexes and cleavage.";
RL J. Neurochem. 72:327-337(1999).
RN [20]
RP FUNCTION (BOTULINUM NEUROTOXIN A LIGHT CHAIN), ACTIVE SITE, CATALYTIC
RP ACTIVITY, BIOPHYSICOCHEMICAL PROPERTIES, AND MUTAGENESIS OF GLU-224.
RC STRAIN=Type A;
RX PubMed=10694409; DOI=10.1021/bi992321x;
RA Li L., Binz T., Niemann H., Singh B.R.;
RT "Probing the mechanistic role of glutamate residue in the zinc-binding
RT motif of type A botulinum neurotoxin light chain.";
RL Biochemistry 39:2399-2405(2000).
RN [21]
RP BIOTECHNOLOGY.
RC STRAIN=NCTC 2916 / Type A;
RX PubMed=10768948; DOI=10.1128/iai.68.5.2587-2593.2000;
RA Chaddock J.A., Purkiss J.R., Friis L.M., Broadbridge J.D., Duggan M.J.,
RA Fooks S.J., Shone C.C., Quinn C.P., Foster K.A.;
RT "Inhibition of vesicular secretion in both neuronal and nonneuronal cells
RT by a retargeted endopeptidase derivative of Clostridium botulinum
RT neurotoxin type A.";
RL Infect. Immun. 68:2587-2593(2000).
RN [22]
RP FUNCTION (BOTULINUM NEUROTOXIN A LIGHT CHAIN), COFACTOR, AND MUTAGENESIS OF
RP GLU-262; PHE-266 AND TYR-366.
RC STRAIN=P64 / Type A;
RX PubMed=11700044; DOI=10.1006/bbrc.2001.5911;
RA Rigoni M., Caccin P., Johnson E.A., Montecucco C., Rossetto O.;
RT "Site-directed mutagenesis identifies active-site residues of the light
RT chain of botulinum neurotoxin type A.";
RL Biochem. Biophys. Res. Commun. 288:1231-1237(2001).
RN [23]
RP FUNCTION (BOTULINUM NEUROTOXIN A LIGHT CHAIN), BIOPHYSICOCHEMICAL
RP PROPERTIES, AND MUTAGENESIS OF GLU-351; ARG-363 AND TYR-366.
RC STRAIN=62A / P64 / Type A;
RX PubMed=11827515; DOI=10.1021/bi0157969;
RA Binz T., Bade S., Rummel A., Kollewe A., Alves J.;
RT "Arg(362) and Tyr(365) of the botulinum neurotoxin type a light chain are
RT involved in transition state stabilization.";
RL Biochemistry 41:1717-1723(2002).
RN [24]
RP REQUIREMENT FOR GANGLIOSIDES FOR ACTIVITY (BOTULINUM NEUROTOXIN TYPE A).
RC STRAIN=Type A;
RX PubMed=12089155; DOI=10.1074/jbc.m205258200;
RA Yowler B.C., Kensinger R.D., Schengrund C.L.;
RT "Botulinum neurotoxin A activity is dependent upon the presence of specific
RT gangliosides in neuroblastoma cells expressing synaptotagmin I.";
RL J. Biol. Chem. 277:32815-32819(2002).
RN [25]
RP FUNCTION (BOTULINUM NEUROTOXIN A HEAVY CHAIN), GANGLIOSIDE-BINDING
RP (BOTULINUM NEUROTOXIN A HEAVY CHAIN), MUTAGENESIS OF GLU-1203; HIS-1253;
RP SER-1264; TRP-1266 AND TYR-1267, AND LIPID-BINDING.
RC STRAIN=62A / Type A;
RX PubMed=14731268; DOI=10.1046/j.1365-2958.2003.03872.x;
RA Rummel A., Mahrhold S., Bigalke H., Binz T.;
RT "The HCC-domain of botulinum neurotoxins A and B exhibits a singular
RT ganglioside binding site displaying serotype specific carbohydrate
RT interaction.";
RL Mol. Microbiol. 51:631-643(2004).
RN [26]
RP IDENTIFICATION OF HOST RECEPTOR (BOTULINUM NEUROTOXIN TYPE A AND BOTULINUM
RP NEUROTOXIN A HEAVY CHAIN), AND INTERACTION WITH HOST SV2C.
RC STRAIN=Type A;
RX PubMed=16545378; DOI=10.1016/j.febslet.2006.02.074;
RA Mahrhold S., Rummel A., Bigalke H., Davletov B., Binz T.;
RT "The synaptic vesicle protein 2C mediates the uptake of botulinum
RT neurotoxin A into phrenic nerves.";
RL FEBS Lett. 580:2011-2014(2006).
RN [27]
RP IDENTIFICATION OF HOST RECEPTOR (BOTULINUM NEUROTOXIN TYPE A), AND
RP INTERACTION WITH HOST SV2 PROTEINS.
RC STRAIN=Type A;
RX PubMed=16543415; DOI=10.1126/science.1123654;
RA Dong M., Yeh F., Tepp W.H., Dean C., Johnson E.A., Janz R., Chapman E.R.;
RT "SV2 is the protein receptor for botulinum neurotoxin A.";
RL Science 312:592-596(2006).
RN [28]
RP FUNCTION (BOTULINUM NEUROTOXIN A LIGHT AND HEAVY CHAIN), DOMAIN, AND
RP DISULFIDE BOND.
RX PubMed=17666397; DOI=10.1074/jbc.m703619200;
RA Fischer A., Montal M.;
RT "Crucial role of the disulfide bridge between botulinum neurotoxin light
RT and heavy chains in protease translocation across membranes.";
RL J. Biol. Chem. 282:29604-29611(2007).
RN [29]
RP DISCUSSION OF BELT FUNCTION (BOTULINUM NEUROTOXIN A HEAVY CHAIN), AND
RP DOMAIN.
RX PubMed=17907800; DOI=10.1371/journal.ppat.0030113;
RA Brunger A.T., Breidenbach M.A., Jin R., Fischer A., Santos J.S., Montal M.;
RT "Botulinum neurotoxin heavy chain belt as an intramolecular chaperone for
RT the light chain.";
RL PLoS Pathog. 3:1191-1194(2007).
RN [30]
RP IDENTIFICATION OF HOST RECEPTOR (BOTULINUM NEUROTOXIN TYPE A), AND
RP INTERACTION WITH HOST SV2 PROTEINS.
RC STRAIN=Type A;
RX PubMed=18815274; DOI=10.1091/mbc.e08-07-0765;
RA Dong M., Liu H., Tepp W.H., Johnson E.A., Janz R., Chapman E.R.;
RT "Glycosylated SV2A and SV2B mediate the entry of botulinum neurotoxin E
RT into neurons.";
RL Mol. Biol. Cell 19:5226-5237(2008).
RN [31]
RP FUNCTION (BOTULINUM NEUROTOXIN A HEAVY CHAIN), AND DOMAIN.
RC STRAIN=Type A;
RX PubMed=19096517; DOI=10.1371/journal.ppat.1000245;
RA Fischer A., Mushrush D.J., Lacy D.B., Montal M.;
RT "Botulinum neurotoxin devoid of receptor binding domain translocates active
RT protease.";
RL PLoS Pathog. 4:E1000245-E1000245(2008).
RN [32]
RP FUNCTION (BOTULINUM NEUROTOXIN A HEAVY CHAIN), DOMAIN, AND
RP PHOSPHATIDYLINOSITOL-BINDING.
RC STRAIN=Type A;
RX PubMed=19161982; DOI=10.1016/j.bbrc.2009.01.037;
RA Muraro L., Tosatto S., Motterlini L., Rossetto O., Montecucco C.;
RT "The N-terminal half of the receptor domain of botulinum neurotoxin A binds
RT to microdomains of the plasma membrane.";
RL Biochem. Biophys. Res. Commun. 380:76-80(2009).
RN [33]
RP FUNCTION (BOTULINUM NEUROTOXIN TYPE A), FUNCTION (BOTULINUM NEUROTOXIN A
RP HEAVY CHAIN), INTERACTION WITH HOST SV2 PROTEINS, AND MUTAGENESIS OF
RP TRP-1266.
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 [34]
RP FUNCTION (BOTULINUM NEUROTOXIN TYPE A), AND SUBCELLULAR LOCATION (BOTULINUM
RP NEUROTOXIN A HEAVY CHAIN).
RC STRAIN=Type A;
RX PubMed=21632541; DOI=10.1074/jbc.m111.254086;
RA Kroken A.R., Karalewitz A.P., Fu Z., Kim J.J., Barbieri J.T.;
RT "Novel ganglioside-mediated entry of botulinum neurotoxin serotype D into
RT neurons.";
RL J. Biol. Chem. 286:26828-26837(2011).
RN [35]
RP FUNCTION (BOTULINUM NEUROTOXIN TYPE A), SUBCELLULAR LOCATION (BOTULINUM
RP NEUROTOXIN A HEAVY CHAIN), AND ACTIVITY REGULATION.
RX PubMed=21832053; DOI=10.1074/jbc.m111.283879;
RA Harper C.B., Martin S., Nguyen T.H., Daniels S.J., Lavidis N.A.,
RA Popoff M.R., Hadzic G., Mariana A., Chau N., McCluskey A., Robinson P.J.,
RA Meunier F.A.;
RT "Dynamin inhibition blocks botulinum neurotoxin type A endocytosis in
RT neurons and delays botulism.";
RL J. Biol. Chem. 286:35966-35976(2011).
RN [36]
RP FUNCTION (BOTULINUM NEUROTOXIN A HEAVY CHAIN), TRANSCYTOSIS ACROSS HOST
RP CELLS, BIOTECHNOLOGY, AND MUTAGENESIS OF HIS-1253; 1266-TRP-TYR-1267;
RP TRP-1266 AND TYR-1267.
RC STRAIN=62A / Type A;
RX PubMed=21106906; DOI=10.1124/jpet.110.175018;
RA Elias M., Al-Saleem F., Ancharski D.M., Singh A., Nasser Z., Olson R.M.,
RA Simpson L.L.;
RT "Evidence that botulinum toxin receptors on epithelial cells and neuronal
RT cells are not identical: implications for development of a non-neurotropic
RT vaccine.";
RL J. Pharmacol. Exp. Ther. 336:605-612(2011).
RN [37]
RP FUNCTION (BOTULINUM NEUROTOXIN A HEAVY CHAIN), AND DOMAIN.
RC STRAIN=Type A;
RX PubMed=22158863; DOI=10.1074/jbc.c111.319400;
RA Fischer A., Sambashivan S., Brunger A.T., Montal M.;
RT "Beltless translocation domain of botulinum neurotoxin A embodies a minimum
RT ion-conductive channel.";
RL J. Biol. Chem. 287:1657-1661(2012).
RN [38]
RP FUNCTION (BOTULINUM NEUROTOXIN A HEAVY CHAIN), AND SUBCELLULAR LOCATION
RP (BOTULINUM NEUROTOXIN A HEAVY CHAIN).
RX PubMed=23471747; DOI=10.1007/s12035-013-8423-9;
RA Colasante C., Rossetto O., Morbiato L., Pirazzini M., Molgo J.,
RA Montecucco C.;
RT "Botulinum neurotoxin type A is internalized and translocated from small
RT synaptic vesicles at the neuromuscular junction.";
RL Mol. Neurobiol. 48:120-127(2013).
RN [39]
RP POSSIBLE IDENTIFICATION OF HOST RECEPTOR (BOTULINUM NEUROTOXIN TYPE A).
RC STRAIN=Type A;
RX PubMed=23696738; DOI=10.1371/journal.ppat.1003369;
RA Jacky B.P., Garay P.E., Dupuy J., Nelson J.B., Cai B., Molina Y., Wang J.,
RA Steward L.E., Broide R.S., Francis J., Aoki K.R., Stevens R.C.,
RA Fernandez-Salas E.;
RT "Identification of fibroblast growth factor receptor 3 (FGFR3) as a protein
RT receptor for botulinum neurotoxin serotype A (BoNT/A).";
RL PLoS Pathog. 9:E1003369-E1003369(2013).
RN [40]
RP RECOGNITION OF RECEPTOR (BOTULINUM NEUROTOXIN A HEAVY CHAIN), AND SUBUNIT.
RC STRAIN=62A / Type A;
RX PubMed=27313224; DOI=10.1042/bcj20160439;
RA Mahrhold S., Bergstroem T., Stern D., Dorner B.G., Aastot C., Rummel A.;
RT "Only the complex N559-glycan in the synaptic vesicle glycoprotein 2C
RT mediates high affinity binding to botulinum neurotoxin serotype A1.";
RL Biochem. J. 473:2645-2654(2016).
RN [41]
RP UBIQUITINATION (BOTULINUM NEUROTOXIN A LIGHT CHAIN).
RX PubMed=28584101; DOI=10.1073/pnas.1621076114;
RA Tsai Y.C., Kotiya A., Kiris E., Yang M., Bavari S., Tessarollo L.,
RA Oyler G.A., Weissman A.M.;
RT "Deubiquitinating enzyme VCIP135 dictates the duration of botulinum
RT neurotoxin type A intoxication.";
RL Proc. Natl. Acad. Sci. U.S.A. 114:E5158-E5166(2017).
RN [42]
RP REVIEW.
RX PubMed=23518040; DOI=10.1016/j.toxicon.2013.02.014;
RA Simpson L.;
RT "The life history of a botulinum toxin molecule.";
RL Toxicon 68:40-59(2013).
RN [43]
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 [44] {ECO:0007744|PDB:1XTF, ECO:0007744|PDB:1XTG}
RP X-RAY CRYSTALLOGRAPHY (2.10 ANGSTROMS) OF 2-420 IN COMPLEX WITH ZINC WITH
RP AND WITHOUT SUBSTRATE, FUNCTION (BOTULINUM NEUROTOXIN A LIGHT CHAIN),
RP SUBSTRATE SPECIFICITY, AND COFACTOR.
RC STRAIN=Type A;
RX PubMed=15592454; DOI=10.1038/nature03123;
RA Breidenbach M.A., Brunger A.T.;
RT "Substrate recognition strategy for botulinum neurotoxin serotype A.";
RL Nature 432:925-929(2004).
RN [45]
RP STRUCTURE BY ELECTRON MICROSCOPY (24.0 ANGSTROMS), AND DOMAIN.
RC STRAIN=Type A;
RX PubMed=18032388; DOI=10.1074/jbc.m707917200;
RA Fischer A., Garcia-Rodriguez C., Geren I., Lou J., Marks J.D., Nakagawa T.,
RA Montal M.;
RT "Molecular architecture of botulinum neurotoxin E revealed by single
RT particle electron microscopy.";
RL J. Biol. Chem. 283:3997-4003(2008).
RN [46] {ECO:0007744|PDB:2VU9, ECO:0007744|PDB:2VUA}
RP X-RAY CRYSTALLOGRAPHY (1.60 ANGSTROMS) OF 874-1296 IN COMPLEX WITH
RP GANGLIOSIDE GT1B ANALOG, FUNCTION (BOTULINUM NEUROTOXIN A HEAVY CHAIN), AND
RP GANGLIOSIDE-BINDING.
RC STRAIN=Type A;
RX PubMed=18704164; DOI=10.1371/journal.ppat.1000129;
RA Stenmark P., Dupuy J., Imamura A., Kiso M., Stevens R.C.;
RT "Crystal structure of botulinum neurotoxin type A in complex with the cell
RT surface co-receptor GT1b-insight into the toxin-neuron interaction.";
RL PLoS Pathog. 4:e1000129-e1000129(2008).
RN [47] {ECO:0007744|PDB:2W2D}
RP X-RAY CRYSTALLOGRAPHY (2.59 ANGSTROMS) OF 1-445 AND 447-877 IN COMPLEX WITH
RP ZINC, FUNCTION (BOTULINUM NEUROTOXIN A LIGHT AND HEAVY CHAIN), COFACTOR,
RP DOMAIN, AND DISULFIDE BONDS.
RC STRAIN=NCTC 2916 / Type A;
RX PubMed=19351593; DOI=10.1016/j.bbrc.2009.02.003;
RA Masuyer G., Thiyagarajan N., James P.L., Marks P.M., Chaddock J.A.,
RA Acharya K.R.;
RT "Crystal structure of a catalytically active, non-toxic endopeptidase
RT derivative of Clostridium botulinum toxin A.";
RL Biochem. Biophys. Res. Commun. 381:50-53(2009).
RN [48] {ECO:0007744|PDB:3V0A, ECO:0007744|PDB:3V0B, ECO:0007744|PDB:3V0C}
RP X-RAY CRYSTALLOGRAPHY (2.70 ANGSTROMS) IN COMPLEX WITH ZINC AND NTNHA,
RP FUNCTION (BOTULINUM NEUROTOXIN A HEAVY CHAIN), COFACTOR, SUBUNIT, DOMAIN,
RP DISULFIDE BONDS, AND MUTAGENESIS OF 861-ARG--LYS-871; 862-LEU--THR-867;
RP GLU-982; LYS-1000; ASP-1037; ASP-1118 AND ASP-1171.
RC STRAIN=Type A;
RX PubMed=22363010; DOI=10.1126/science.1214270;
RA Gu S., Rumpel S., Zhou J., Strotmeier J., Bigalke H., Perry K.,
RA Shoemaker C.B., Rummel A., Jin R.;
RT "Botulinum neurotoxin is shielded by NTNHA in an interlocked complex.";
RL Science 335:977-981(2012).
RN [49] {ECO:0007744|PDB:4JRA}
RP X-RAY CRYSTALLOGRAPHY (2.30 ANGSTROMS) OF 871-1296 IN COMPLEX WITH SV2C
RP RECEPTOR FRAGMENT, FUNCTION (BOTULINUM NEUROTOXIN A HEAVY CHAIN), SUBUNIT,
RP SUBCELLULAR LOCATION (BOTULINUM NEUROTOXIN A HEAVY CHAIN), AND MUTAGENESIS
RP OF 1145-THR-THR-1146; ARG-1156 AND ARG-1294.
RC STRAIN=Type A;
RX PubMed=24240280; DOI=10.1038/nature12732;
RA Benoit R.M., Frey D., Hilbert M., Kevenaar J.T., Wieser M.M.,
RA Stirnimann C.U., McMillan D., Ceska T., Lebon F., Jaussi R.,
RA Steinmetz M.O., Schertler G.F., Hoogenraad C.C., Capitani G.,
RA Kammerer R.A.;
RT "Structural basis for recognition of synaptic vesicle protein 2C by
RT botulinum neurotoxin A.";
RL Nature 505:108-111(2014).
RN [50] {ECO:0007744|PDB:5JLV, ECO:0007744|PDB:5JMC}
RP X-RAY CRYSTALLOGRAPHY (2.00 ANGSTROMS) OF 872-1296 IN COMPLEX WITH HUMAN
RP AND SV2C RECEPTOR, FUNCTION (BOTULINUM NEUROTOXIN TYPE A AND BOTULINUM
RP NEUROTOXIN A HEAVY CHAIN), SV2 GLYCAN-BINDING, AND MUTAGENESIS OF PHE-953;
RP HIS-1064; 1145-THR-THR-1146; ARG-1156; GLY-1292 AND ARG-1294.
RC STRAIN=Type A;
RX PubMed=27294781; DOI=10.1038/nsmb.3245;
RA Yao G., Zhang S., Mahrhold S., Lam K.H., Stern D., Bagramyan K., Perry K.,
RA Kalkum M., Rummel A., Dong M., Jin R.;
RT "N-linked glycosylation of SV2 is required for binding and uptake of
RT botulinum neurotoxin A.";
RL Nat. Struct. Mol. Biol. 23:656-662(2016).
RN [51] {ECO:0007744|PDB:5TPB, ECO:0007744|PDB:5TPC}
RP X-RAY CRYSTALLOGRAPHY (2.00 ANGSTROMS) OF 876-1296 IN COMPLEX WITH
RP GANGLIOSIDE ANALOG, AND FUNCTION (BOTULINUM NEUROTOXIN A HEAVY CHAIN).
RC STRAIN=Type A;
RX PubMed=27958736; DOI=10.1021/jacs.6b09534;
RA Hamark C., Berntsson R.P., Masuyer G., Henriksson L.M., Gustafsson R.,
RA Stenmark P., Widmalm G.;
RT "Glycans confer specificity to the recognition of ganglioside receptors by
RT botulinum neurotoxin A.";
RL J. Am. Chem. Soc. 139:218-230(2017).
RN [52] {ECO:0007744|PDB:5MK6, ECO:0007744|PDB:5MK7}
RP X-RAY CRYSTALLOGRAPHY (1.45 ANGSTROMS) OF 871-1296, AND DISULFIDE BOND.
RX PubMed=29576992; DOI=10.7717/peerj.4552;
RA Davies J.R., Hackett G.S., Liu S.M., Acharya K.R.;
RT "High resolution crystal structures of the receptor-binding domain of
RT Clostridium botulinum neurotoxin serotypes A and FA.";
RL PeerJ 6:E4552-E4552(2018).
CC -!- FUNCTION: [Botulinum neurotoxin type A]: 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:15394302, PubMed:7578132). Precursor of
CC botulinum neurotoxin A which has 2 coreceptors; complex polysialylated
CC gangliosides found on neural tissue and specific membrane-anchored
CC proteins of synaptic vesicles. Receptor proteins are exposed on host
CC presynaptic cell membrane during neurotransmitter release, when the
CC toxin heavy chain (HC) binds to them. Upon synaptic vesicle recycling
CC the toxin is taken up via the endocytic pathway. When the pH of the
CC toxin-containing endosome drops a structural rearrangement occurs so
CC that the N-terminus of the HC forms pores that allows the light chain
CC (LC) to translocate into the cytosol (PubMed:17666397,
CC PubMed:19096517). Once in the cytosol the disulfide bond linking the 2
CC subunits is reduced and LC cleaves its target protein on synaptic
CC vesicles, preventing their fusion with the cytoplasmic membrane and
CC thus neurotransmitter release. Toxin activity requires polysialylated
CC gangliosides; GT1b supports activity better than GD1a
CC (PubMed:12089155). Binds to host peripheral neuronal presynaptic
CC membranes via the synaptic vesicle glycoproteins SV2A, SV2B and SV2C
CC (PubMed:16543415). It binds directly to the largest lumenal
CC (intravesicular) loop of SV2A, SV2B and SV2C that is transiently
CC exposed outside of cells during exocytosis; gangliosides enhance
CC binding (PubMed:16543415, PubMed:16545378, PubMed:18815274). Recognizes
CC an N-linked glycan on SV2 proteins (PubMed:18815274, PubMed:27294781).
CC May also use FGFR3 as a receptor (PubMed:23696738). Toxin uptake into
CC neural cells requires stimulation (incubation with K(+) to stimulate
CC receptor exposure) to be internalized by receptor-mediated endocytosis
CC (PubMed:16543415, PubMed:19650874, PubMed:21632541, PubMed:21832053).
CC Subsequently the toxin colocalizes with its receptor in host cells
CC (PubMed:16543415, PubMed:19650874). Toxin uptake can be blocked by the
CC appropriate SV2 protein fragments in cell culture (PubMed:16543415).
CC {ECO:0000269|PubMed:12089155, ECO:0000269|PubMed:15394302,
CC ECO:0000269|PubMed:16543415, ECO:0000269|PubMed:16545378,
CC ECO:0000269|PubMed:17666397, ECO:0000269|PubMed:18815274,
CC ECO:0000269|PubMed:19096517, ECO:0000269|PubMed:19650874,
CC ECO:0000269|PubMed:21632541, ECO:0000269|PubMed:21832053,
CC ECO:0000269|PubMed:23696738, ECO:0000269|PubMed:27294781,
CC ECO:0000269|PubMed:7578132}.
CC -!- FUNCTION: [Botulinum neurotoxin A light chain]: Has proteolytic
CC activity (PubMed:7578132). After translocation into the eukaryotic host
CC cytosol LC hydrolyzes the '197-Gln-|-Arg-198' bond in SNAP25, blocking
CC neurotransmitter release (PubMed:8243676, PubMed:7578132,
CC PubMed:9886085, PubMed:10694409, PubMed:11700044, PubMed:11827515,
CC PubMed:19351593). Recognizes the '146-Met--Gly-155' region of SNAP25,
CC which confers substrate specificity (PubMed:9886085, PubMed:15592454).
CC Hydrolyzes the '202-Thr-|-Arg-203' bond of mouse SNAP23, but not in
CC human which has a different sequence (PubMed:9886085). Reduction of the
CC interchain disulfide bond occurs in the host cytosol and probably
CC prevents retrotranslocation into the synaptic vesicle
CC (PubMed:17666397). Has slow (occurs over 4 weeks) autocatalytic
CC cleavage, however it is not clear if this is physiologically relevant
CC (PubMed:11565902). {ECO:0000269|PubMed:10694409,
CC ECO:0000269|PubMed:11565902, ECO:0000269|PubMed:11700044,
CC ECO:0000269|PubMed:11827515, ECO:0000269|PubMed:15592454,
CC ECO:0000269|PubMed:17666397, ECO:0000269|PubMed:7578132,
CC ECO:0000269|PubMed:8243676, ECO:0000269|PubMed:9886085,
CC ECO:0000305|PubMed:19351593}.
CC -!- FUNCTION: [Botulinum neurotoxin A heavy chain]: Responsible for host
CC epithelial cell transcytosis, host nerve cell targeting and
CC translocation of botulinum neurotoxin A light chain (LC) into host
CC cytosol. Composed of 3 subdomains; the translocation domain (TD), and
CC N-terminus and C-terminus of the receptor-binding domain (RBD)
CC (PubMed:19096517). The RBD is responsible for binding to host
CC epithelial cells and transcytosis across them; this uses different
CC receptors than those on nerve cells (PubMed:21106906). RBD is also
CC responsible for adherence of toxin to host nerve cell surface; HC alone
CC prevents uptake of whole toxin by neural cells, and delays paralysis
CC onset by 75% (PubMed:6694738, PubMed:10413679). Isolated RBD also
CC delays paralysis onset (PubMed:21106906). The N-terminus of the RBD
CC binds to phosphatidylinositol, which might play a role in membrane-
CC binding (PubMed:19161982). Binds to host protein receptor synaptic
CC vesicle glycoproteins SV2A, SV2B and SV2C via lumenal loop 4
CC (PubMed:16545378, PubMed:6370252, PubMed:27294781, PubMed:24240280,
CC PubMed:19650874, PubMed:27313224). Binding can be inhibited by protein
CC fragments from either the HC or SV2C (PubMed:24240280). Isolated HC
CC significantly decreases uptake and toxicity of whole BoNT/A, but also
CC interferes with uptake of BoNT/E and to a lesser extent BoNT/F
CC (PubMed:19650874). The RBD recognizes the N-linked glycan on 'Asn-559'
CC of SV2A, SV2B and SV2C; hydrogen-bonding occurs via 10 well-defined
CC water molecules and stacking of hydrophobic residues (PubMed:27294781).
CC Binds one host GT1b ganglioside, which serves as a coreceptor
CC (PubMed:14731268, PubMed:18704164, PubMed:27958736). Modeling shows the
CC HC can bind both coreceptors (a ganglioside and SV2 protein)
CC simultaneously at different sites (PubMed:24240280). Crystals of the
CC RBD with a GT1b analog can be grown at pH 5.5, indicating the toxin-
CC ganglioside complex could be stable within the endosome
CC (PubMed:18704164). Isolated RBD binds NTNHA (a bacterial protein that
CC protects toxin) with high affinity at pH 6.0 but not at pH 7.5
CC (PubMed:22363010). The N-terminal belt (residues 449-545) wraps around
CC the perimeter of the LC, probably protecting Zn(2+) in the active site;
CC it is not required for channel formation by the TD domain but may serve
CC to prevent premature LC dissociation from the translocation channel and
CC to protect toxin prior to translocation (PubMed:22158863,
CC PubMed:17907800, PubMed:19351593). The isolated TD forms transmembrane
CC channels of about 15 Angstroms in the absence of a pH gradient; LC
CC translocation requires a pH and redox gradient (pH 5.0/oxidizing in the
CC cis compartment, pH 7.0/reducing in the trans compartment), LC does not
CC unfold unless the cis pH is 6.0 or less (PubMed:2446925,
CC PubMed:17666397, PubMed:19096517). Pores are presumably made by 1-2
CC toxin molecules (PubMed:23471747). While interaction with the RBD
CC modulates the pH threshold for membrane insertion, the RBD is not
CC essential for toxin degradation of SNAP25 in neural cells
CC (PubMed:19096517). {ECO:0000269|PubMed:10413679,
CC ECO:0000269|PubMed:14731268, ECO:0000269|PubMed:16545378,
CC ECO:0000269|PubMed:17666397, ECO:0000269|PubMed:18704164,
CC ECO:0000269|PubMed:19096517, ECO:0000269|PubMed:19161982,
CC ECO:0000269|PubMed:19351593, ECO:0000269|PubMed:19650874,
CC ECO:0000269|PubMed:21106906, ECO:0000269|PubMed:22158863,
CC ECO:0000269|PubMed:22363010, ECO:0000269|PubMed:23471747,
CC ECO:0000269|PubMed:24240280, ECO:0000269|PubMed:27294781,
CC ECO:0000269|PubMed:27313224, ECO:0000269|PubMed:27958736,
CC ECO:0000269|PubMed:6694738, ECO:0000305|PubMed:17907800,
CC ECO:0000305|PubMed:2446925}.
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:0000269|PubMed:10694409, ECO:0000269|PubMed:7578132,
CC ECO:0000269|PubMed:8243676, ECO:0000269|PubMed:9886085};
CC -!- COFACTOR:
CC Name=Zn(2+); Xref=ChEBI:CHEBI:29105;
CC Evidence={ECO:0000269|PubMed:11565902, ECO:0000269|PubMed:11700044,
CC ECO:0000269|PubMed:1429690, ECO:0000269|PubMed:15592454,
CC ECO:0000269|PubMed:19351593, ECO:0000269|PubMed:22363010,
CC ECO:0000269|PubMed:7578132};
CC Note=Binds 1 zinc ion per subunit (PubMed:1429690, PubMed:11700044,
CC PubMed:15592454, PubMed:19351593, PubMed:22363010).
CC {ECO:0000269|PubMed:11700044, ECO:0000269|PubMed:1429690,
CC ECO:0000269|PubMed:15592454, ECO:0000269|PubMed:19351593,
CC ECO:0000269|PubMed:22363010};
CC -!- ACTIVITY REGULATION: Toxin internalization is inhibited by azide or
CC dinitrophenol or at 4 degrees Celsius (PubMed:6694738). Dynamin (DNM)
CC inhibitors abolish toxin uptake (PubMed:21832053).
CC {ECO:0000269|PubMed:21832053, ECO:0000269|PubMed:6694738}.
CC -!- BIOPHYSICOCHEMICAL PROPERTIES:
CC Kinetic parameters:
CC KM=41 uM for purified SNAP25 with isolated botulinum neurotoxin A
CC light chain {ECO:0000269|PubMed:10694409};
CC KM=9.8 uM for purified SNAP25 with isolated botulinum neurotoxin A
CC light chain {ECO:0000269|PubMed:11827515};
CC Note=kcat is 140 min(-1) (PubMed:10694409). kcat is 1026 (-1)
CC (PubMed:11827515). {ECO:0000269|PubMed:10694409,
CC ECO:0000269|PubMed:11827515};
CC -!- SUBUNIT: Heterodimer; disulfide-linked heterodimer of a light chain
CC (LC) and heavy chain (HC) (PubMed:7578132). Interacts with glycosylated
CC host synaptic vesicle glycoproteins SV2A, SV2B and SV2C which serve as
CC coreceptors (PubMed:16543415, PubMed:18815274, PubMed:19650874,
CC PubMed:24240280, PubMed:27313224). Glycosylation of 'Asn-559' in SV2C
CC contributes a 12-fold increase in affinity to this interaction
CC (PubMed:27313224). Depolarization of target tissue with high levels of
CC K(+) leads to greater levels of receptor exposure (PubMed:16543415). In
CC vitro addition of gangliosides increases SV2-toxin interaction
CC (PubMed:16543415). Forms a highly interlocked heterodimer with NTNHA at
CC pH 6.0 but not at pH 7.5 called the minimally functional progenitor
CC toxin complex (M-PTC) (PubMed:22363010). The PTC is thought to protect
CC toxin in the host acidic gastrointestinal tract, facilitate
CC transcytosis across the intestinal barrier and release at neutral pH as
CC is found in the bloodstream (PubMed:22363010).
CC {ECO:0000269|PubMed:16543415, ECO:0000269|PubMed:16545378,
CC ECO:0000269|PubMed:18815274, ECO:0000269|PubMed:19650874,
CC ECO:0000269|PubMed:22363010, ECO:0000269|PubMed:24240280,
CC ECO:0000269|PubMed:27313224, ECO:0000269|PubMed:7578132,
CC ECO:0000305|PubMed:22363010}.
CC -!- INTERACTION:
CC P0DPI0; Q02563: Sv2a; Xeno; NbExp=2; IntAct=EBI-8178893, EBI-466194;
CC P0DPI0; Q496J9: SV2C; Xeno; NbExp=4; IntAct=EBI-8178893, EBI-16081469;
CC P0DPI0; Q9Z2I6: Sv2c; Xeno; NbExp=12; IntAct=EBI-8178893, EBI-8178859;
CC -!- SUBCELLULAR LOCATION: [Botulinum neurotoxin type A]: Secreted
CC {ECO:0000269|PubMed:7592120}. Secreted, cell wall
CC {ECO:0000269|PubMed:7592120}. Host synapse, host presynaptic cell
CC membrane {ECO:0000269|PubMed:6694738}. Note=Whole toxin may be released
CC from the bacteria during cell wall exfoliation (PubMed:7592120). There
CC are estimated to be 150-500 toxin molecules per um(2) of non-myelinated
CC mouse hemidiaphragm nerve membrane (PubMed:6694738). In mouse
CC hemidiaphragm binds only to nerve terminals, and not to muscle, blood
CC vessels, connective tissue Schwann cells or myelin, toxin can be
CC internalized by this preparation (PubMed:6694738).
CC {ECO:0000269|PubMed:6694738, ECO:0000269|PubMed:7592120}.
CC -!- SUBCELLULAR LOCATION: [Botulinum neurotoxin A light chain]: Secreted
CC {ECO:0000269|PubMed:6370252}. Host cytoplasm, host cytosol
CC {ECO:0000305|PubMed:7578132, ECO:0000305|PubMed:8243676,
CC ECO:0000305|PubMed:9886085}.
CC -!- SUBCELLULAR LOCATION: [Botulinum neurotoxin A heavy chain]: Secreted
CC {ECO:0000269|PubMed:6370252}. Host synapse, host presynaptic cell
CC membrane {ECO:0000269|PubMed:6694738, ECO:0000305|PubMed:10413679}.
CC Host cytoplasmic vesicle, host secretory vesicle, host synaptic vesicle
CC membrane {ECO:0000269|PubMed:23471747, ECO:0000305|PubMed:10413679,
CC ECO:0000305|PubMed:21632541, ECO:0000305|PubMed:21832053,
CC ECO:0000305|PubMed:24240280, ECO:0000305|PubMed:6694738}; Multi-pass
CC membrane protein {ECO:0000305|PubMed:17666397,
CC ECO:0000305|PubMed:19096517, ECO:0000305|PubMed:2446925}. Note=Whole
CC toxin may be released from the bacteria during cell wall exfoliation
CC (PubMed:7592120). Colocalizes with its receptor SV2C (synaptic vesicle
CC glycoprotein 2C) and VGAT (vesicular inhibitory amino acid transporter)
CC in neurons (PubMed:24240280). In neurons HC colocalizes with
CC synaptophysin or VAMP2 probably in synaptic vesicles, a portion also
CC colocalizes with RAB5 and may be in synaptic vesicle protein sorting
CC endosomes (PubMed:21632541, PubMed:21832053). Therefore there may be
CC more than one uptake pathway at nerve terminals. Uptake of HC and whole
CC toxin is slowed by dynamin inhibitors (PubMed:21832053). 1-2 molecules
CC of HC are found in the host synaptic vesicle lumen, uptake and
CC subsequent release of LC is very rapid (PubMed:21832053,
CC PubMed:23471747). {ECO:0000269|PubMed:21632541,
CC ECO:0000269|PubMed:21832053, ECO:0000269|PubMed:23471747,
CC ECO:0000269|PubMed:24240280}.
CC -!- INDUCTION: In cultured bacteria, first detected in late exponential
CC growth (17 hours), reaches maximal levels at 24-25 hours and remains
CC nearly constant for 5 days (at protein level).
CC {ECO:0000269|PubMed:7592120}.
CC -!- DOMAIN: [Botulinum neurotoxin A light chain]: Has protease activity
CC (PubMed:7578132). {ECO:0000269|PubMed:7578132}.
CC -!- DOMAIN: [Botulinum neurotoxin A 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). Upon trypsin digestion the isolated TD forms channels in
CC bilayers when the cis side is acidic/oxidizing and the trans side is pH
CC 7.0/reducing (PubMed:2446925, PubMed:17666397, PubMed:19096517). The
CC RBD rotates 140 degrees around the TD in the presence of NTNHA
CC (PubMed:22363010). The 3 major domains each serve as a chaperone for
CC the other 2 to ensure they act only in the correct host cell context
CC (PubMed:19096517). In BoNT/A structures the LC is separated from the
CC RBD by the TD; the belt wraps around the perimeter of the LC,
CC protecting Zn(2+) in the active site (PubMed:18032388, PubMed:19351593,
CC PubMed:22363010). The belt region (449-545) may be a pseudosubstrate
CC inhibitor which serves as an intramolecular chaperone for the LC prior
CC to its translocation into the host cytosol (PubMed:17907800).
CC {ECO:0000269|PubMed:17666397, ECO:0000269|PubMed:18032388,
CC ECO:0000269|PubMed:19096517, ECO:0000269|PubMed:19351593,
CC ECO:0000269|PubMed:22363010, ECO:0000269|PubMed:2446925,
CC ECO:0000305|PubMed:17907800}.
CC -!- PTM: In a bacterial culture the precursor chain is initally cleaved on
CC the amino side of Gly-445 and is processed more slowly between Lys-448
CC and Ala-449 to give the final mature heavy chain sequence.
CC {ECO:0000269|PubMed:2126206}.
CC -!- PTM: [Botulinum neurotoxin A light chain]: Has slow autocatalytic
CC activity, cleaves 250-Tyr-Tyr-251, 266-Phe-Gly-267, 419-Phe-Thr-420,
CC 423-Phe-Glu-424, 430-Cys-Val-431, 432-Arg-Gly-433, 438-Lys-Thr-439, and
CC probably 429-Leu-Cys-430 over a period of 4 weeks. Catalysis of the
CC '197-Gln-|-Arg-198' bond in SNAP25 is estimated to be 10(5) more
CC efficient than autocatalysis, leaving the physiological importance of
CC autocatalysis in doubt (PubMed:11565902).
CC {ECO:0000269|PubMed:11565902}.
CC -!- PTM: [Botulinum neurotoxin A light chain]: Ubiquitinated by host HECD2.
CC Deubiquitination by host VCPIP1 prevents degradation by the proteasome.
CC {ECO:0000269|PubMed:28584101}.
CC -!- BIOTECHNOLOGY: Dynamin inhibitors slow toxin uptake by nerve cells, and
CC might be used to prolong the treatment window for antitoxins.
CC {ECO:0000305|PubMed:21832053}.
CC -!- BIOTECHNOLOGY: Replacement of the RBD by other proteins (such as wheat
CC germ agglutinin) allows the rest of the toxin to be taken up by other
CC cell types, and can be used for investigating synaptic vesicle docking-
CC dependent processes in BoNT resistant cells (PubMed:10768948). LC
CC retains protease activity (PubMed:10768948, PubMed:19351593).
CC {ECO:0000269|PubMed:10768948, ECO:0000269|PubMed:19351593}.
CC -!- BIOTECHNOLOGY: Isolated receptor-binding domain (RBD) can be used as a
CC vaccine; a mutated form that is transcytosed into the general
CC circulation but does not enter nerve cells (Leu-1266-1267-Ser) is as
CC efficient as wild-type RBD (PubMed:21106906).
CC {ECO:0000269|PubMed:21106906}.
CC -!- PHARMACEUTICAL: Available under the name Botox (onabotulinumtoxinA,
CC Allergan), Dysport (abobotulinumtoxinA, Ipsen Biopharmaceuticals) and
CC Xeomin (incobotulinumtoxinA, Merz Pharmaceuticals) for the treatment of
CC strabismus and blepharospasm associated with dystonia and cervical
CC dystonia. Also used for the treatment of hemifacial spasm and a number
CC of other neurological disorders characterized by abnormal muscle
CC contraction. It is also used cosmetically to smooth facial wrinkles.
CC {ECO:0000305|PubMed:28356439}.
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. {ECO:0000305}.
CC -!- MISCELLANEOUS: Types A, B and E are the most frequent cause of adult
CC human foodborne botulism; type A is the most severe, while type E has
CC the shortest incubation period (PubMed:1431246).
CC {ECO:0000269|PubMed:1431246}.
CC -!- MISCELLANEOUS: Neurotoxin type A is released from bacteria in the two-
CC chain form (PubMed:6370252, PubMed:2126206).
CC {ECO:0000269|PubMed:2126206, ECO:0000269|PubMed:6370252}.
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:0000305|PubMed:21106906}.
CC -!- SIMILARITY: Belongs to the peptidase M27 family. {ECO:0000305}.
CC -!- CAUTION: Contradictory results show that only SV2C is the receptor; in
CC these experiments gangliosides do not improve toxin-coreceptor
CC interaction (PubMed:16545378). {ECO:0000269|PubMed:16545378}.
CC -!- WEB RESOURCE: Name=BOTOX product information Web site;
CC URL="https://www.botox.com/";
CC -!- WEB RESOURCE: Name=Protein Spotlight; Note=From sausages to wrinkles
CC - Issue 19 of February 2002;
CC URL="https://web.expasy.org/spotlight/back_issues/019";
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; X52066; CAA36289.1; -; Genomic_DNA.
DR EMBL; M30196; AAA23262.1; -; Genomic_DNA.
DR EMBL; X92973; CAA63551.1; -; Genomic_DNA.
DR EMBL; D67030; BAA11051.1; -; Genomic_DNA.
DR PIR; A35294; BTCLAB.
DR RefSeq; WP_003356619.1; NZ_LFOS01000037.1.
DR RefSeq; WP_011948511.1; NZ_LHUL01000004.1.
DR PDB; 1XTF; X-ray; 2.20 A; A/B=2-420.
DR PDB; 1XTG; X-ray; 2.10 A; A=2-420.
DR PDB; 2ILP; X-ray; 1.90 A; A/B=3-424.
DR PDB; 2IMA; X-ray; 1.94 A; A/B=1-424.
DR PDB; 2IMB; X-ray; 2.41 A; A/B=3-424.
DR PDB; 2IMC; X-ray; 2.00 A; A/B=3-424.
DR PDB; 2ISE; X-ray; 2.20 A; A/B=1-420.
DR PDB; 2ISG; X-ray; 2.00 A; A/B=1-420.
DR PDB; 2ISH; X-ray; 2.00 A; A/B=1-420.
DR PDB; 2VU9; X-ray; 1.60 A; A=876-1296.
DR PDB; 2VUA; X-ray; 1.70 A; A=876-1296.
DR PDB; 2W2D; X-ray; 2.59 A; A/C=3-442, B/D=447-877.
DR PDB; 3BOK; X-ray; 1.25 A; A=3-425.
DR PDB; 3BON; X-ray; 1.20 A; A=3-425.
DR PDB; 3BOO; X-ray; 1.40 A; A=3-425.
DR PDB; 3V0A; X-ray; 2.70 A; A=1-1296.
DR PDB; 3V0B; X-ray; 3.90 A; A=1-1296.
DR PDB; 3V0C; X-ray; 4.30 A; A=1-1296.
DR PDB; 3ZUR; X-ray; 2.71 A; A/B=3-430, A/B=454-865.
DR PDB; 3ZUS; X-ray; 2.95 A; A/B/C/D=3-431, A/B/C/D=454-865.
DR PDB; 4HEV; X-ray; 2.50 A; A/B=1-425.
DR PDB; 4IQP; X-ray; 2.30 A; A=871-1296.
DR PDB; 4JRA; X-ray; 2.30 A; A/B=871-1296.
DR PDB; 5JLV; X-ray; 2.00 A; A/B=872-1296.
DR PDB; 5JMC; X-ray; 2.64 A; A/C/E/G=872-1296.
DR PDB; 5MK6; X-ray; 1.45 A; A=871-1296.
DR PDB; 5MK7; X-ray; 1.80 A; A=871-1296.
DR PDB; 5TPB; X-ray; 2.60 A; A/B=876-1296.
DR PDB; 5TPC; X-ray; 2.00 A; A=876-1296.
DR PDB; 5V8P; X-ray; 2.50 A; A/B=1-424.
DR PDB; 5V8R; X-ray; 1.90 A; A/B=1-424.
DR PDB; 5V8U; X-ray; 2.05 A; A/B=1-424.
DR PDB; 5VGV; X-ray; 2.60 A; A=1-425.
DR PDB; 5VGX; X-ray; 2.15 A; A=1-425.
DR PDB; 6DKK; X-ray; 2.70 A; A/B=547-871.
DR PDB; 6MHJ; X-ray; 3.02 A; A=547-871.
DR PDB; 6XCB; X-ray; 1.74 A; A=3-424.
DR PDB; 6XCC; X-ray; 1.90 A; A=3-424.
DR PDB; 6XCD; X-ray; 1.92 A; A=3-424.
DR PDB; 6XCE; X-ray; 2.50 A; A=3-424.
DR PDB; 6XCF; X-ray; 1.68 A; A=3-424.
DR PDB; 7KYF; X-ray; 2.33 A; C=1-417.
DR PDB; 7KYH; X-ray; 2.91 A; A/B/C/D=1-417.
DR PDB; 7LZP; X-ray; 2.86 A; A/D=2-438.
DR PDB; 7M1H; X-ray; 2.78 A; A=2-438.
DR PDBsum; 1XTF; -.
DR PDBsum; 1XTG; -.
DR PDBsum; 2ILP; -.
DR PDBsum; 2IMA; -.
DR PDBsum; 2IMB; -.
DR PDBsum; 2IMC; -.
DR PDBsum; 2ISE; -.
DR PDBsum; 2ISG; -.
DR PDBsum; 2ISH; -.
DR PDBsum; 2VU9; -.
DR PDBsum; 2VUA; -.
DR PDBsum; 2W2D; -.
DR PDBsum; 3BOK; -.
DR PDBsum; 3BON; -.
DR PDBsum; 3BOO; -.
DR PDBsum; 3V0A; -.
DR PDBsum; 3V0B; -.
DR PDBsum; 3V0C; -.
DR PDBsum; 3ZUR; -.
DR PDBsum; 3ZUS; -.
DR PDBsum; 4HEV; -.
DR PDBsum; 4IQP; -.
DR PDBsum; 4JRA; -.
DR PDBsum; 5JLV; -.
DR PDBsum; 5JMC; -.
DR PDBsum; 5MK6; -.
DR PDBsum; 5MK7; -.
DR PDBsum; 5TPB; -.
DR PDBsum; 5TPC; -.
DR PDBsum; 5V8P; -.
DR PDBsum; 5V8R; -.
DR PDBsum; 5V8U; -.
DR PDBsum; 5VGV; -.
DR PDBsum; 5VGX; -.
DR PDBsum; 6DKK; -.
DR PDBsum; 6MHJ; -.
DR PDBsum; 6XCB; -.
DR PDBsum; 6XCC; -.
DR PDBsum; 6XCD; -.
DR PDBsum; 6XCE; -.
DR PDBsum; 6XCF; -.
DR PDBsum; 7KYF; -.
DR PDBsum; 7KYH; -.
DR PDBsum; 7LZP; -.
DR PDBsum; 7M1H; -.
DR AlphaFoldDB; P0DPI0; -.
DR SMR; P0DPI0; -.
DR IntAct; P0DPI0; 4.
DR MINT; P0DPI0; -.
DR BindingDB; P0DPI0; -.
DR ChEMBL; CHEMBL5192; -.
DR UniLectin; P0DPI0; -.
DR ABCD; P0DPI0; 61 sequenced antibodies.
DR GeneID; 5185061; -.
DR Reactome; R-HSA-5250968; Toxicity of botulinum toxin type A (botA).
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:0016021; C:integral component of membrane; IEA:UniProtKB-KW.
DR GO; GO:1905576; F:ganglioside GT1b binding; IDA:UniProtKB.
DR GO; GO:0004222; F:metalloendopeptidase activity; IEA:UniProtKB-EC.
DR GO; GO:0008320; F:protein transmembrane transporter activity; EXP:Reactome.
DR GO; GO:0090729; F:toxin activity; IEA:UniProtKB-KW.
DR GO; GO:0008270; F:zinc ion binding; IDA:UniProtKB.
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; Cell wall; Direct protein sequencing; Disulfide bond;
KW Host cell membrane; Host cytoplasm; Host cytoplasmic vesicle;
KW Host membrane; Host synapse; Hydrolase; Lipid-binding; Membrane;
KW Metal-binding; Metalloprotease; Neurotoxin; Pharmaceutical; Protease;
KW Secreted; Toxin; Transmembrane; Transmembrane helix; Ubl conjugation;
KW Virulence; Zinc.
FT INIT_MET 1
FT /note="Removed"
FT /evidence="ECO:0000269|PubMed:2126206,
FT ECO:0000269|PubMed:6370252, ECO:0000269|Ref.6,
FT ECO:0000305|PubMed:11565902"
FT CHAIN 2..1296
FT /note="Botulinum neurotoxin type A"
FT /id="PRO_0000444902"
FT CHAIN 2..448
FT /note="Botulinum neurotoxin A light chain"
FT /evidence="ECO:0000269|PubMed:6370252"
FT /id="PRO_0000029211"
FT CHAIN 449..1296
FT /note="Botulinum neurotoxin A heavy chain"
FT /evidence="ECO:0000269|PubMed:6370252"
FT /id="PRO_0000029212"
FT TRANSMEM 627..647
FT /note="Helical"
FT /evidence="ECO:0000255"
FT TRANSMEM 656..676
FT /note="Helical"
FT /evidence="ECO:0000255"
FT REGION 449..870
FT /note="Translocation domain (TD); not required to bind
FT NTNHA"
FT /evidence="ECO:0000269|PubMed:22363010,
FT ECO:0000305|PubMed:19096517"
FT REGION 492..545
FT /note="Belt; not required for channel formation"
FT /evidence="ECO:0000305|PubMed:22158863"
FT REGION 871..1092
FT /note="N-terminus of receptor binding domain (N-RBD)"
FT /evidence="ECO:0000305|PubMed:19096517"
FT REGION 1093..1296
FT /note="C-terminus of receptor binding domain (C-RBD)"
FT /evidence="ECO:0000305|PubMed:19096517"
FT REGION 1252..1253
FT /note="Interaction with host ganglioside GT1b"
FT /evidence="ECO:0000269|PubMed:18704164,
FT ECO:0000269|PubMed:27958736, ECO:0000305|PubMed:14731268,
FT ECO:0007744|PDB:2VU9, ECO:0007744|PDB:5TPB,
FT ECO:0007744|PDB:5TPC"
FT MOTIF 1264..1267
FT /note="Host ganglioside-binding motif; interacts with GT1b"
FT /evidence="ECO:0000269|PubMed:18704164,
FT ECO:0000269|PubMed:27958736, ECO:0000305|PubMed:14731268,
FT ECO:0000305|PubMed:19650874, ECO:0007744|PDB:2VU9,
FT ECO:0007744|PDB:5TPB, ECO:0007744|PDB:5TPC"
FT ACT_SITE 224
FT /note="Proton acceptor"
FT /evidence="ECO:0000255|PROSITE-ProRule:PRU10095,
FT ECO:0000305|PubMed:10694409"
FT BINDING 223
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:15592454, ECO:0000269|PubMed:19351593,
FT ECO:0007744|PDB:1XTF, ECO:0007744|PDB:1XTG,
FT ECO:0007744|PDB:2W2D, ECO:0007744|PDB:3V0A,
FT ECO:0007744|PDB:3V0B"
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:15592454, ECO:0000269|PubMed:19351593,
FT ECO:0000305|PubMed:7578132, ECO:0007744|PDB:1XTF,
FT ECO:0007744|PDB:1XTG, ECO:0007744|PDB:2W2D,
FT ECO:0007744|PDB:3V0A, ECO:0007744|PDB:3V0B"
FT BINDING 262
FT /ligand="Zn(2+)"
FT /ligand_id="ChEBI:CHEBI:29105"
FT /ligand_note="catalytic"
FT /evidence="ECO:0000269|PubMed:15592454,
FT ECO:0000269|PubMed:19351593, ECO:0000305|PubMed:11700044,
FT ECO:0007744|PDB:1XTF, ECO:0007744|PDB:1XTG,
FT ECO:0007744|PDB:2W2D, ECO:0007744|PDB:3V0A,
FT ECO:0007744|PDB:3V0B"
FT BINDING 1117
FT /ligand="ganglioside GT1b (d18:1(4E))"
FT /ligand_id="ChEBI:CHEBI:78452"
FT /ligand_note="host ganglioside"
FT /evidence="ECO:0000269|PubMed:18704164,
FT ECO:0000269|PubMed:27958736, ECO:0007744|PDB:2VU9,
FT ECO:0007744|PDB:5TPB, ECO:0007744|PDB:5TPC"
FT BINDING 1203
FT /ligand="ganglioside GT1b (d18:1(4E))"
FT /ligand_id="ChEBI:CHEBI:78452"
FT /ligand_note="host ganglioside"
FT /evidence="ECO:0000269|PubMed:18704164,
FT ECO:0000269|PubMed:27958736, ECO:0000305|PubMed:14731268,
FT ECO:0007744|PDB:2VU9, ECO:0007744|PDB:5TPB,
FT ECO:0007744|PDB:5TPC"
FT SITE 363
FT /note="Transition state stabilizer"
FT /evidence="ECO:0000305|PubMed:11827515"
FT SITE 366
FT /note="Transition state stabilizer"
FT /evidence="ECO:0000305|PubMed:11827515"
FT DISULFID 430..454
FT /note="Interchain (between light and heavy chains)"
FT /evidence="ECO:0000269|PubMed:19351593,
FT ECO:0000269|PubMed:22363010, ECO:0000305|PubMed:17666397,
FT ECO:0007744|PDB:2W2D, ECO:0007744|PDB:3V0A,
FT ECO:0007744|PDB:3V0B, ECO:0007744|PDB:3V0C"
FT DISULFID 1235..1280
FT /evidence="ECO:0007744|PDB:3V0C, ECO:0007744|PDB:5MK7"
FT VARIANT 27
FT /note="V -> A (in strain: 62A)"
FT MUTAGEN 224
FT /note="E->D: Light chain has 5% cleavage activity on
FT SNAP25. KM for SNAP25 is nearly wild-type."
FT /evidence="ECO:0000269|PubMed:10694409"
FT MUTAGEN 224
FT /note="E->Q: Light chain no longer cleaves SNAP25, no
FT effect on substrate or Zn(2+) binding."
FT /evidence="ECO:0000269|PubMed:10694409"
FT MUTAGEN 227
FT /note="H->Y: Light chain no longer cleaves SNAP25, not
FT toxic in vitro or in vivo when reconstituted with heavy
FT chain."
FT /evidence="ECO:0000269|PubMed:7578132"
FT MUTAGEN 262
FT /note="E->A: Light chain has 20% cleavage activity on
FT SNAP25, 40% decrease in Zn(2+)."
FT /evidence="ECO:0000269|PubMed:11700044"
FT MUTAGEN 266
FT /note="F->A: Light chain has 50% cleavage activity on
FT SNAP25, no effect on Zn(2+) binding."
FT /evidence="ECO:0000269|PubMed:11700044"
FT MUTAGEN 351
FT /note="E->A,Q: Wild-type KM for SNAP25, no protease
FT activity, about 30% less Zn(2+)."
FT /evidence="ECO:0000269|PubMed:11827515"
FT MUTAGEN 363
FT /note="R->A,H,K: Wild-type KM for SNAP25, about 75-fold
FT decrease in kcat, no effect on Zn(2+) binding."
FT /evidence="ECO:0000269|PubMed:11827515"
FT MUTAGEN 366
FT /note="Y->A: Light chain has 40% cleavage activity on
FT SNAP25, 30% decrease in Zn(2+)."
FT /evidence="ECO:0000269|PubMed:11700044"
FT MUTAGEN 366
FT /note="Y->F: About wild-type KM for SNAP25, 35-fold
FT decrease in kcat, no effect on Zn(2+) binding."
FT /evidence="ECO:0000269|PubMed:11827515"
FT MUTAGEN 861..871
FT /note="RLLSTFTEYIK->ALLSTFTPYIP: Reduced toxicity."
FT /evidence="ECO:0000269|PubMed:22363010"
FT MUTAGEN 862..867
FT /note="LLSTFT->KESTFK: Reduced toxicity."
FT /evidence="ECO:0000269|PubMed:22363010"
FT MUTAGEN 953
FT /note="F->G: Whole toxin has 50-fold reduction in toxicity,
FT almost no binding of RBD to neurons."
FT /evidence="ECO:0000269|PubMed:27294781"
FT MUTAGEN 953
FT /note="F->R: Whole toxin is non-toxic, almost no binding of
FT RBD to neurons."
FT /evidence="ECO:0000269|PubMed:27294781"
FT MUTAGEN 982
FT /note="E->A,Q: Decreased binding of NTNHA by receptor-
FT binding domain (RBD) at pH 7.5."
FT /evidence="ECO:0000269|PubMed:22363010"
FT MUTAGEN 1000
FT /note="K->A: Decreased binding of NTNHA by RBD at pH 6.0,
FT none at pH 7.5."
FT /evidence="ECO:0000269|PubMed:22363010"
FT MUTAGEN 1037
FT /note="D->A,N: Decreased binding of NTNHA by RBD at pH
FT 7.5."
FT /evidence="ECO:0000269|PubMed:22363010"
FT MUTAGEN 1064
FT /note="H->G,R: Whole toxin has reduced toxicity,
FT dramatically reduced binding of RBD to neurons."
FT /evidence="ECO:0000269|PubMed:27294781"
FT MUTAGEN 1118
FT /note="D->A: Decreased binding of NTNHA by RBD at pH 7.5."
FT /evidence="ECO:0000269|PubMed:22363010"
FT MUTAGEN 1145..1146
FT /note="TT->AA: No binding of RBD to neurons. Loss of
FT binding to SV2C."
FT /evidence="ECO:0000269|PubMed:24240280,
FT ECO:0000269|PubMed:27294781"
FT MUTAGEN 1156
FT /note="R->E: Decreased binding of RBD to SV2C, substantial
FT binding to neurons."
FT /evidence="ECO:0000269|PubMed:24240280,
FT ECO:0000269|PubMed:27294781"
FT MUTAGEN 1171
FT /note="D->A: Decreased binding of NTNHA by RBD at pH 7.5."
FT /evidence="ECO:0000269|PubMed:22363010"
FT MUTAGEN 1203
FT /note="E->L: Strongly reduced toxicity, heavy chain has
FT very strongly reduced binding to synaptosomes, decreased
FT binding to gangioside GT1b."
FT /evidence="ECO:0000269|PubMed:14731268"
FT MUTAGEN 1253
FT /note="H->A: Strongly reduced toxicity, heavy chain has
FT very strongly reduced binding to synaptosomes, decrease in
FT ganglioside GT1b binding."
FT /evidence="ECO:0000269|PubMed:14731268"
FT MUTAGEN 1253
FT /note="H->W: Heavy chain has very strongly reduced binding
FT to synaptosomes, binds much less GT1b. RBD protects against
FT neurotoxin less well than wild-type."
FT /evidence="ECO:0000269|PubMed:14731268,
FT ECO:0000269|PubMed:21106906"
FT MUTAGEN 1264
FT /note="S->A: Reduced toxicity, heavy chain has strongly
FT reduced binding to synaptosomes, heavy chain binds less
FT GT1b."
FT /evidence="ECO:0000269|PubMed:14731268"
FT MUTAGEN 1266..1267
FT /note="WY->LS: Whole RBD does not protect against
FT neurotoxin, no effect on epithelial cell passage; can be
FT used as a vaccine."
FT /evidence="ECO:0000269|PubMed:21106906"
FT MUTAGEN 1266
FT /note="W->L: Nearly complete loss of toxicity, heavy chain
FT has very strongly reduced binding to synaptosomes, binds
FT much less GT1b. Heavy chain no longer inhibits whole-toxin
FT uptake and toxicity. RBD protects against neurotoxin
FT considerably less well than wild-type."
FT /evidence="ECO:0000269|PubMed:14731268,
FT ECO:0000269|PubMed:19650874, ECO:0000269|PubMed:21106906"
FT MUTAGEN 1267
FT /note="Y->F: Nearly complete loss of toxicity, heavy chain
FT has very strongly reduced binding to synaptosomes, binds
FT much less GT1b."
FT /evidence="ECO:0000269|PubMed:14731268"
FT MUTAGEN 1267
FT /note="Y->S: Nearly complete loss of toxicity, heavy chain
FT has very strongly reduced binding to synaptosomes, binds
FT much less GT1b. RBD protects against neurotoxin
FT considerably less well than wild-type."
FT /evidence="ECO:0000269|PubMed:14731268,
FT ECO:0000269|PubMed:21106906"
FT MUTAGEN 1292
FT /note="G->Q: Whole toxin has very strongly reduced
FT toxicity, almost no binding of RBD to neurons."
FT /evidence="ECO:0000269|PubMed:27294781"
FT MUTAGEN 1292
FT /note="G->R: Nearly complete loss of toxicity, almost no
FT binding of RBD to neurons."
FT /evidence="ECO:0000269|PubMed:27294781"
FT MUTAGEN 1294
FT /note="R->A: Decreased binding of RBD to SV2C, substantial
FT binding to neurons."
FT /evidence="ECO:0000269|PubMed:24240280"
FT MUTAGEN 1294
FT /note="R->S: Decreased binding of RBD to SV2C, substantial
FT binding to neurons."
FT /evidence="ECO:0000269|PubMed:27294781"
FT CONFLICT 2
FT /note="P -> Q (in Ref. 1; CAA36289)"
FT /evidence="ECO:0000305"
FT CONFLICT 480
FT /note="E -> P (in Ref. 9; AA sequence)"
FT /evidence="ECO:0000305"
FT STRAND 10..12
FT /evidence="ECO:0007829|PDB:7KYH"
FT STRAND 16..23
FT /evidence="ECO:0007829|PDB:6XCF"
FT TURN 25..27
FT /evidence="ECO:0007829|PDB:6XCF"
FT STRAND 33..39
FT /evidence="ECO:0007829|PDB:6XCF"
FT STRAND 42..48
FT /evidence="ECO:0007829|PDB:6XCF"
FT STRAND 51..53
FT /evidence="ECO:0007829|PDB:7KYF"
FT HELIX 54..56
FT /evidence="ECO:0007829|PDB:7LZP"
FT HELIX 64..66
FT /evidence="ECO:0007829|PDB:6XCB"
FT TURN 68..70
FT /evidence="ECO:0007829|PDB:7KYF"
FT TURN 75..78
FT /evidence="ECO:0007829|PDB:6XCF"
FT HELIX 81..99
FT /evidence="ECO:0007829|PDB:6XCF"
FT HELIX 102..113
FT /evidence="ECO:0007829|PDB:6XCF"
FT STRAND 126..128
FT /evidence="ECO:0007829|PDB:6XCF"
FT HELIX 131..133
FT /evidence="ECO:0007829|PDB:6XCF"
FT STRAND 134..138
FT /evidence="ECO:0007829|PDB:6XCF"
FT STRAND 144..148
FT /evidence="ECO:0007829|PDB:6XCF"
FT STRAND 150..155
FT /evidence="ECO:0007829|PDB:6XCF"
FT STRAND 157..161
FT /evidence="ECO:0007829|PDB:7KYH"
FT STRAND 164..166
FT /evidence="ECO:0007829|PDB:6XCF"
FT TURN 175..177
FT /evidence="ECO:0007829|PDB:6XCF"
FT STRAND 184..187
FT /evidence="ECO:0007829|PDB:6XCF"
FT STRAND 192..196
FT /evidence="ECO:0007829|PDB:6XCF"
FT HELIX 200..203
FT /evidence="ECO:0007829|PDB:6XCF"
FT STRAND 206..208
FT /evidence="ECO:0007829|PDB:7M1H"
FT HELIX 217..232
FT /evidence="ECO:0007829|PDB:6XCF"
FT STRAND 242..244
FT /evidence="ECO:0007829|PDB:6XCF"
FT HELIX 249..253
FT /evidence="ECO:0007829|PDB:6XCF"
FT STRAND 257..259
FT /evidence="ECO:0007829|PDB:6XCF"
FT HELIX 260..266
FT /evidence="ECO:0007829|PDB:6XCF"
FT HELIX 268..273
FT /evidence="ECO:0007829|PDB:6XCF"
FT HELIX 276..299
FT /evidence="ECO:0007829|PDB:6XCF"
FT STRAND 302..308
FT /evidence="ECO:0007829|PDB:6XCF"
FT HELIX 310..321
FT /evidence="ECO:0007829|PDB:6XCF"
FT STRAND 323..325
FT /evidence="ECO:0007829|PDB:7LZP"
FT STRAND 327..329
FT /evidence="ECO:0007829|PDB:6XCD"
FT STRAND 331..333
FT /evidence="ECO:0007829|PDB:7LZP"
FT HELIX 335..346
FT /evidence="ECO:0007829|PDB:6XCF"
FT HELIX 351..358
FT /evidence="ECO:0007829|PDB:6XCF"
FT STRAND 366..368
FT /evidence="ECO:0007829|PDB:6XCF"
FT STRAND 372..375
FT /evidence="ECO:0007829|PDB:6XCF"
FT TURN 381..383
FT /evidence="ECO:0007829|PDB:6XCF"
FT TURN 386..388
FT /evidence="ECO:0007829|PDB:6XCF"
FT STRAND 393..395
FT /evidence="ECO:0007829|PDB:6XCE"
FT HELIX 396..398
FT /evidence="ECO:0007829|PDB:7KYF"
FT TURN 399..401
FT /evidence="ECO:0007829|PDB:7KYH"
FT HELIX 402..404
FT /evidence="ECO:0007829|PDB:6XCF"
FT TURN 406..409
FT /evidence="ECO:0007829|PDB:6XCF"
FT HELIX 410..412
FT /evidence="ECO:0007829|PDB:6XCF"
FT STRAND 414..418
FT /evidence="ECO:0007829|PDB:6XCF"
FT TURN 420..422
FT /evidence="ECO:0007829|PDB:6XCE"
FT STRAND 428..431
FT /evidence="ECO:0007829|PDB:7LZP"
FT STRAND 548..550
FT /evidence="ECO:0007829|PDB:6DKK"
FT HELIX 551..555
FT /evidence="ECO:0007829|PDB:6DKK"
FT STRAND 562..564
FT /evidence="ECO:0007829|PDB:6DKK"
FT STRAND 567..570
FT /evidence="ECO:0007829|PDB:6DKK"
FT HELIX 572..576
FT /evidence="ECO:0007829|PDB:6DKK"
FT STRAND 581..583
FT /evidence="ECO:0007829|PDB:6DKK"
FT HELIX 588..594
FT /evidence="ECO:0007829|PDB:6DKK"
FT TURN 600..602
FT /evidence="ECO:0007829|PDB:6DKK"
FT HELIX 603..619
FT /evidence="ECO:0007829|PDB:6DKK"
FT STRAND 622..625
FT /evidence="ECO:0007829|PDB:6DKK"
FT STRAND 627..633
FT /evidence="ECO:0007829|PDB:6DKK"
FT STRAND 636..639
FT /evidence="ECO:0007829|PDB:6DKK"
FT HELIX 643..645
FT /evidence="ECO:0007829|PDB:6DKK"
FT STRAND 646..649
FT /evidence="ECO:0007829|PDB:6DKK"
FT STRAND 653..657
FT /evidence="ECO:0007829|PDB:6DKK"
FT STRAND 662..668
FT /evidence="ECO:0007829|PDB:6DKK"
FT HELIX 679..685
FT /evidence="ECO:0007829|PDB:6MHJ"
FT HELIX 688..719
FT /evidence="ECO:0007829|PDB:6DKK"
FT HELIX 721..751
FT /evidence="ECO:0007829|PDB:6DKK"
FT HELIX 756..760
FT /evidence="ECO:0007829|PDB:6DKK"
FT HELIX 766..798
FT /evidence="ECO:0007829|PDB:6DKK"
FT HELIX 800..824
FT /evidence="ECO:0007829|PDB:6DKK"
FT TURN 827..832
FT /evidence="ECO:0007829|PDB:6DKK"
FT HELIX 834..845
FT /evidence="ECO:0007829|PDB:6DKK"
FT HELIX 853..856
FT /evidence="ECO:0007829|PDB:6DKK"
FT HELIX 861..868
FT /evidence="ECO:0007829|PDB:6DKK"
SQ SEQUENCE 1296 AA; 149454 MW; B731EF5BA5E62FDA CRC64;
MPFVNKQFNY KDPVNGVDIA YIKIPNVGQM QPVKAFKIHN KIWVIPERDT FTNPEEGDLN
PPPEAKQVPV SYYDSTYLST DNEKDNYLKG VTKLFERIYS TDLGRMLLTS IVRGIPFWGG
STIDTELKVI DTNCINVIQP DGSYRSEELN LVIIGPSADI IQFECKSFGH EVLNLTRNGY
GSTQYIRFSP DFTFGFEESL EVDTNPLLGA GKFATDPAVT LAHELIHAGH RLYGIAINPN
RVFKVNTNAY YEMSGLEVSF EELRTFGGHD AKFIDSLQEN EFRLYYYNKF KDIASTLNKA
KSIVGTTASL QYMKNVFKEK YLLSEDTSGK FSVDKLKFDK LYKMLTEIYT EDNFVKFFKV
LNRKTYLNFD KAVFKINIVP KVNYTIYDGF NLRNTNLAAN FNGQNTEINN MNFTKLKNFT
GLFEFYKLLC VRGIITSKTK SLDKGYNKAL NDLCIKVNNW DLFFSPSEDN FTNDLNKGEE
ITSDTNIEAA EENISLDLIQ QYYLTFNFDN EPENISIENL SSDIIGQLEL MPNIERFPNG
KKYELDKYTM FHYLRAQEFE HGKSRIALTN SVNEALLNPS RVYTFFSSDY VKKVNKATEA
AMFLGWVEQL VYDFTDETSE VSTTDKIADI TIIIPYIGPA LNIGNMLYKD DFVGALIFSG
AVILLEFIPE IAIPVLGTFA LVSYIANKVL TVQTIDNALS KRNEKWDEVY KYIVTNWLAK
VNTQIDLIRK KMKEALENQA EATKAIINYQ YNQYTEEEKN NINFNIDDLS SKLNESINKA
MININKFLNQ CSVSYLMNSM IPYGVKRLED FDASLKDALL KYIYDNRGTL IGQVDRLKDK
VNNTLSTDIP FQLSKYVDNQ RLLSTFTEYI KNIINTSILN LRYESNHLID LSRYASKINI
GSKVNFDPID KNQIQLFNLE SSKIEVILKN AIVYNSMYEN FSTSFWIRIP KYFNSISLNN
EYTIINCMEN NSGWKVSLNY GEIIWTLQDT QEIKQRVVFK YSQMINISDY INRWIFVTIT
NNRLNNSKIY INGRLIDQKP ISNLGNIHAS NNIMFKLDGC RDTHRYIWIK YFNLFDKELN
EKEIKDLYDN QSNSGILKDF WGDYLQYDKP YYMLNLYDPN KYVDVNNVGI RGYMYLKGPR
GSVMTTNIYL NSSLYRGTKF IIKKYASGNK DNIVRNNDRV YINVVVKNKE YRLATNASQA
GVEKILSALE IPDVGNLSQV VVMKSKNDQG ITNKCKMNLQ DNNGNDIGFI GFHQFNNIAK
LVASNWYNRQ IERSSRTLGC SWEFIPVDDG WGERPL
