A5HZZ9 (BXA1_CLOBH) Reviewed, UniProtKB/Swiss-Prot

Names and origin

Protein names

Recommended name:
Botulinum neurotoxin type A
Short name=BoNT/A
EC=3.4.24.69

Alternative name(s):
Bontoxilysin-A
Short name=BOTOX

Cleaved into the following 2 chains:

1. Botulinum neurotoxin A light chain
2. Botulinum neurotoxin A heavy chain

Gene names

Name:	botA
Ordered Locus Names:	CBO0806, CLC_0862

Organism

Clostridium botulinum (strain Hall / ATCC 3502 / NCTC 13319 / Type A) [Reference proteome] [HAMAP]

Taxonomic identifier

441771 [NCBI]

Taxonomic lineage

cellular organisms › Bacteria › Firmicutes › Clostridia › Clostridiales › Clostridiaceae › Clostridium › Clostridium botulinum › Clostridium botulinum A

Protein attributes

Sequence length	1296 AA.
Sequence status	Complete.
Sequence processing	The displayed sequence is further processed into a mature form.
Protein existence	Evidence at protein level

General annotation (Comments)

Function	Inhibits acetylcholine release. The botulinum toxin binds with high affinity to peripheral neuronal presynaptic membrane to the secretory vesicle protein SV2. It binds directly to the largest luminal loop of SV2A, SV2B and SV2C. It is then internalized by receptor-mediated endocytosis. The C-terminus of the heavy chain (H) is responsible for the adherence of the toxin to the cell surface while the N-terminus mediates transport of the light chain from the endocytic vesicle to the cytosol. After translocation, the light chain (L) hydrolyzes the 197-Gln-|-Arg-198 bond in SNAP-25, thereby blocking neurotransmitter release. Inhibition of acetylcholine release results in flaccid paralysis, with frequent heart or respiratory failure.
Catalytic activity	Limited hydrolysis of proteins of the neuroexocytosis apparatus, synaptobrevins, SNAP25 or syntaxin. No detected action on small molecule substrates.
Cofactor	Binds 1 zinc ion per subunit.
Subunit structure	Disulfide-linked heterodimer of a light chain (L) and a heavy chain (H).
Subcellular location	Botulinum neurotoxin A light chain: Secreted. Host cytoplasm › host cytosol. Botulinum neurotoxin A heavy chain: Secreted. Host cell junction › host synapse › host presynaptic cell membrane; Multi-pass membrane protein Potential.
Pharmaceutical use	Available under the name BOTOX (Allergan) for the treatment of strabismus and blepharospasm associated with dystonia and cervical dystonia. Also used for the treatment of hemifacial spasm and a number of other neurological disorders characterized by abnormal muscle contraction.
Miscellaneous	There are seven antigenically distinct forms of botulinum neurotoxin: Types A, B, C1, D, E, F, and G.
Sequence similarities	Belongs to the peptidase M27 family.

Ontologies

Keywords
Biological process	Virulence
Cellular component	Host cell junction Host cell membrane Host cytoplasm Host membrane Host synapse Membrane Secreted
Domain	Transmembrane Transmembrane helix
Ligand	Metal-binding Zinc
Molecular function	Hydrolase Metalloprotease Neurotoxin Protease Toxin
PTM	Disulfide bond
Technical term	3D-structure Complete proteome Direct protein sequencing Pharmaceutical Reference proteome
Gene Ontology (GO)
Biological_process	inhibition of neurotransmitter uptake Inferred from electronic annotation. Source: InterPro pathogenesis Inferred from electronic annotation. Source: UniProtKB-KW proteolysis Inferred from electronic annotation. Source: UniProtKB-KW
Cellular_component	host cell cytosol Inferred from electronic annotation. Source: UniProtKB-SubCell host cell junction Inferred from electronic annotation. Source: UniProtKB-KW host cell presynaptic membrane Inferred from electronic annotation. Source: UniProtKB-SubCell integral to membrane Inferred from electronic annotation. Source: UniProtKB-KW
Molecular_function	metalloendopeptidase activity Inferred from electronic annotation. Source: InterPro zinc ion binding Inferred from electronic annotation. Source: InterPro
Complete GO annotation...

Sequence annotation (Features)

Feature key

Position(s)

Length

Description

Graphical view

Feature identifier

Molecule processing

Initiator methionine

1

1

Removed By similarity

Chain

2 – 448

447

Botulinum neurotoxin A light chain

PRO_0000308906

Chain

449 – 1296

848

Botulinum neurotoxin A heavy chain

PRO_0000308907

Regions

Transmembrane

627 – 647

21

Helical; Potential

Transmembrane

656 – 676

21

Helical; Potential

Sites

Active site

224

1

By similarity

Metal binding

223

1

Zinc; catalytic By similarity

Metal binding

227

1

Zinc; catalytic By similarity

Metal binding

262

1

Zinc; catalytic By similarity

Amino acid modifications

Disulfide bond

430 ↔ 454

Interchain (between light and heavy chains) By similarity

Disulfide bond

1235 ↔ 1280

By similarity

Experimental info

Sequence conflict

1218

1

S → Y AA sequence Ref.4

Secondary structure

1

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1296

Helix Strand Turn

Details...

Beta strand

4 – 6

3

Beta strand

16 – 23

8

Beta strand

26 – 28

3

Beta strand

33 – 39

7

Beta strand

42 – 48

7

Beta strand

51 – 53

3

Helix

54 – 56

3

Beta strand

63 – 65

3

Turn

68 – 70

3

Turn

75 – 78

4

Helix

81 – 99

19

Helix

102 – 113

12

Beta strand

126 – 128

3

Helix

131 – 133

3

Beta strand

134 – 138

5

Beta strand

144 – 148

5

Beta strand

150 – 155

6

Beta strand

164 – 166

3

Turn

175 – 177

3

Beta strand

184 – 187

4

Beta strand

192 – 196

5

Helix

200 – 203

4

Beta strand

211 – 214

4

Helix

217 – 232

16

Beta strand

242 – 244

3

Helix

249 – 252

4

Turn

253 – 255

3

Beta strand

257 – 259

3

Helix

260 – 266

7

Helix

268 – 273

6

Helix

276 – 299

24

Beta strand

302 – 308

7

Helix

310 – 321

12

Beta strand

323 – 325

3

Beta strand

327 – 329

3

Beta strand

331 – 333

3

Helix

335 – 347

13

Helix

351 – 358

8

Beta strand

366 – 368

3

Beta strand

372 – 375

4

Turn

381 – 383

3

Turn

386 – 388

3

Helix

396 – 398

3

Turn

400 – 404

5

Turn

406 – 409

4

Helix

410 – 412

3

Beta strand

414 – 418

5

Turn

420 – 422

3

Sequences

	Sequence		Length	Mass (Da)
	A5HZZ9 [UniParc]. Last modified June 26, 2007. Version 1. Checksum: DEA8CF2754AE43E6	FASTA	1,296	149,426
	10 20 30 40 50 60 MPFVNKQFNY KDPVNGVDIA YIKIPNAGQM QPVKAFKIHN KIWVIPERDT FTNPEEGDLN 70 80 90 100 110 120 PPPEAKQVPV SYYDSTYLST DNEKDNYLKG VTKLFERIYS TDLGRMLLTS IVRGIPFWGG 130 140 150 160 170 180 STIDTELKVI DTNCINVIQP DGSYRSEELN LVIIGPSADI IQFECKSFGH EVLNLTRNGY 190 200 210 220 230 240 GSTQYIRFSP DFTFGFEESL EVDTNPLLGA GKFATDPAVT LAHELIHAGH RLYGIAINPN 250 260 270 280 290 300 RVFKVNTNAY YEMSGLEVSF EELRTFGGHD AKFIDSLQEN EFRLYYYNKF KDIASTLNKA 310 320 330 340 350 360 KSIVGTTASL QYMKNVFKEK YLLSEDTSGK FSVDKLKFDK LYKMLTEIYT EDNFVKFFKV 370 380 390 400 410 420 LNRKTYLNFD KAVFKINIVP KVNYTIYDGF NLRNTNLAAN FNGQNTEINN MNFTKLKNFT 430 440 450 460 470 480 GLFEFYKLLC VRGIITSKTK SLDKGYNKAL NDLCIKVNNW DLFFSPSEDN FTNDLNKGEE 490 500 510 520 530 540 ITSDTNIEAA EENISLDLIQ QYYLTFNFDN EPENISIENL SSDIIGQLEL MPNIERFPNG 550 560 570 580 590 600 KKYELDKYTM FHYLRAQEFE HGKSRIALTN SVNEALLNPS RVYTFFSSDY VKKVNKATEA 610 620 630 640 650 660 AMFLGWVEQL VYDFTDETSE VSTTDKIADI TIIIPYIGPA LNIGNMLYKD DFVGALIFSG 670 680 690 700 710 720 AVILLEFIPE IAIPVLGTFA LVSYIANKVL TVQTIDNALS KRNEKWDEVY KYIVTNWLAK 730 740 750 760 770 780 VNTQIDLIRK KMKEALENQA EATKAIINYQ YNQYTEEEKN NINFNIDDLS SKLNESINKA 790 800 810 820 830 840 MININKFLNQ CSVSYLMNSM IPYGVKRLED FDASLKDALL KYIYDNRGTL IGQVDRLKDK 850 860 870 880 890 900 VNNTLSTDIP FQLSKYVDNQ RLLSTFTEYI KNIINTSILN LRYESNHLID LSRYASKINI 910 920 930 940 950 960 GSKVNFDPID KNQIQLFNLE SSKIEVILKN AIVYNSMYEN FSTSFWIRIP KYFNSISLNN 970 980 990 1000 1010 1020 EYTIINCMEN NSGWKVSLNY GEIIWTLQDT QEIKQRVVFK YSQMINISDY INRWIFVTIT 1030 1040 1050 1060 1070 1080 NNRLNNSKIY INGRLIDQKP ISNLGNIHAS NNIMFKLDGC RDTHRYIWIK YFNLFDKELN 1090 1100 1110 1120 1130 1140 EKEIKDLYDN QSNSGILKDF WGDYLQYDKP YYMLNLYDPN KYVDVNNVGI RGYMYLKGPR 1150 1160 1170 1180 1190 1200 GSVMTTNIYL NSSLYRGTKF IIKKYASGNK DNIVRNNDRV YINVVVKNKE YRLATNASQA 1210 1220 1230 1240 1250 1260 GVEKILSALE IPDVGNLSQV VVMKSKNDQG ITNKCKMNLQ DNNGNDIGFI GFHQFNNIAK 1270 1280 1290 LVASNWYNRQ IERSSRTLGC SWEFIPVDDG WGERPL

References

	« Hide 'large scale' referencesShow 'large scale' references »
[1]	"Genome sequence of a proteolytic (Group I) Clostridium botulinum strain Hall A and comparative analysis of the clostridial genomes." Sebaihia M., Peck M.W., Minton N.P., Thomson N.R., Holden M.T.G., Mitchell W.J., Carter A.T., Bentley S.D., Mason D.R., Crossman L., Paul C.J., Ivens A., Wells-Bennik M.H.J., Davis I.J., Cerdeno-Tarraga A.M., Churcher C., Quail M.A., Chillingworth T. , Feltwell T., Fraser A., Goodhead I., Hance Z., Jagels K., Larke N., Maddison M., Moule S., Mungall K., Norbertczak H., Rabbinowitsch E., Sanders M., Simmonds M., White B., Whithead S., Parkhill J. Genome Res. 17:1082-1092(2007) [PubMed] [Europe PMC] [Abstract] Cited for: NUCLEOTIDE SEQUENCE [LARGE SCALE GENOMIC DNA]. Strain: Hall / ATCC 3502 / NCTC 13319 / Type A.
[2]	"Analysis of the neurotoxin complex genes in Clostridium botulinum A1-A4 and B1 strains: BoNT/A3, /Ba4 and /B1 clusters are located within plasmids." Smith T.J., Hill K.K., Foley B.T., Detter J.C., Munk A.C., Bruce D.C., Doggett N.A., Smith L.A., Marks J.D., Xie G., Brettin T.S. PLoS ONE 2:E1271-E1271(2007) [PubMed] [Europe PMC] [Abstract] Cited for: NUCLEOTIDE SEQUENCE [LARGE SCALE GENOMIC DNA]. Strain: Hall / ATCC 3502 / NCTC 13319 / Type A.
[3]	"Characterization of botulinum type A neurotoxin gene: delineation of the N-terminal encoding region." Betley M.J., Somers E., Dasgupta B.R. Biochem. Biophys. Res. Commun. 162:1388-1395(1989) [PubMed] [Europe PMC] [Abstract] Cited for: NUCLEOTIDE SEQUENCE [GENOMIC DNA] OF 1-35.
[4]	"Botulinum type A neurotoxin digested with pepsin yields 132, 97, 72, 45, 42, and 18 kD fragments." Gimenez J.A., DasGupta B.R. J. Protein Chem. 12:351-363(1993) [PubMed] [Europe PMC] [Abstract] Cited for: PROTEIN SEQUENCE OF 867-880 AND 1148-1219.
[5]	"Proteolysis of SNAP-25 by types E and A botulinal neurotoxins." Binz T., Blasi J., Yamasaki S., Baumeister A., Link E., Suedhof T.C., Jahn R., Niemann H. J. Biol. Chem. 269:1617-1620(1994) [PubMed] [Europe PMC] [Abstract] Cited for: IDENTIFICATION OF SUBSTRATE.
+	Additional computationally mapped references.

Web resources

BOTOX product information Web site

Protein Spotlight

From sausages to wrinkles - Issue 19 of February 2002

BotDB - A Database Resource for Clostridial Neurotoxins

Cross-references

Sequence databases

EMBL
GenBank
DDBJ

AM412317 Genomic DNA. Translation: CAL82360.1.
CP000727 Genomic DNA. Translation: ABS38337.1.

PIR

BTCLAB. A35294.

RefSeq

YP_001253342.1. NC_009495.1.
YP_001386738.1. NC_009698.1.

3D structure databases

PDBe
RCSB PDB
PDBj

Entry	Method	Resolution (Å)	Chain	Positions	PDBsum
1XTF	X-ray	2.20	A/B	2-420	[»]
1XTG	X-ray	2.10	A	2-422	[»]
2W2D	X-ray	2.59	A/C	3-442	[»]
			B/D	447-877	[»]
3DSE	X-ray	1.90	A	1-417	[»]
3K3Q	X-ray	2.60	B	3-250	[»]
			C	251-425	[»]
3QIX	X-ray	2.41	A/B	3-424	[»]
3QIY	X-ray	2.30	A	3-424	[»]
3QIZ	X-ray	2.00	A	3-424	[»]
3QJ0	X-ray	2.30	A	3-424	[»]
3QW5	X-ray	1.60	A	1-424	[»]
3QW6	X-ray	1.60	A	1-424	[»]
3QW7	X-ray	1.50	A	1-424	[»]
3QW8	X-ray	1.60	A	1-424	[»]

ProteinModelPortal

A5HZZ9.

SMR

A5HZZ9. Positions 1-423.

ModBase

Search...

Protein-protein interaction databases

STRING

413999.CBO0806.

Protocols and materials databases

StructuralBiologyKnowledgebase

Search...

Genome annotation databases

EnsemblBacteria

ABS38337; ABS38337; CLC_0862.

GeneID

5185061.
5398487.

KEGG

cbh:CLC_0862.
cbo:CBO0806.

PATRIC

19363717. VBICloBot22612_0802.

Phylogenomic databases

HOGENOM

HOG000234384.

KO

K06011.

OMA

DFWGNYL.

ProtClustDB

CLSK912378.

Enzyme and pathway databases

BioCyc

CBOT413999:GJ72-872-MONOMER.
CBOT441771:GIWX-833-MONOMER.

Family and domain databases

Gene3D

2.60.120.200. 1 hit.
3.90.1240.10. 1 hit.

InterPro

IPR008985. ConA-like_lec_gl_sf.
IPR013320. ConA-like_subgrp.
IPR011065. Kunitz_inhibitor_ST1-like.
IPR000395. Neurotox_Zn_protease.
IPR013104. Toxin_rcpt-bd_C.
IPR012928. Toxin_rcpt-bd_N.
IPR012500. Toxin_trans.
[Graphical view]

Pfam

PF01742. Peptidase_M27. 1 hit.
PF07951. Toxin_R_bind_C. 1 hit.
PF07953. Toxin_R_bind_N. 1 hit.
PF07952. Toxin_trans. 1 hit.
[Graphical view]

PRINTS

PR00760. BONTOXILYSIN.

SUPFAM

SSF49899. ConA_like_lec_gl. 1 hit.
SSF50386. Kunitz_like. 1 hit.

PROSITE

PS00142. ZINC_PROTEASE. 1 hit.
[Graphical view]

ProtoNet

Search...

Other

BindingDB

A5HZZ9.

ChEMBL

CHEMBL5344.

EvolutionaryTrace

A5HZZ9.

Entry information

Entry name

BXA1_CLOBH

Accession

Primary (citable) accession number: A5HZZ9
Secondary accession number(s): A7G1U9

Entry history

Integrated into UniProtKB/Swiss-Prot:	November 13, 2007
Last sequence update:	June 26, 2007
Last modified:	May 1, 2013
This is version 52 of the entry and version 1 of the sequence. [Complete history]

Entry status

Reviewed (UniProtKB/Swiss-Prot)

Annotation program

Prokaryotic Protein Annotation Program