Reviewed, UniProtKB/Swiss-Prot P19321 (BXD_CLOBO)

Names and origin

Protein names

Recommended name:
    Botulinum neurotoxin type D
      Short name=BoNT/D
    EC=3.4.24.69
Alternative name(s):
    Bontoxilysin-D
Cleaved into the following 2 chains:
    1- Recommended name:
            Botulinum neurotoxin D light chain
    2- Recommended name:
            Botulinum neurotoxin D heavy chain

Gene names

Name:

botD

Organism

Clostridium botulinum

Taxonomic identifier

1491 [NCBI]

Taxonomic lineage

Bacteria › Firmicutes › Clostridia › Clostridiales › Clostridiaceae › Clostridium

Protein attributes

Sequence length	1276 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	Botulinum toxin acts by inhibiting neurotransmitter release. It binds to peripheral neuronal synapses, is internalized and moves by retrograde transport up the axon into the spinal cord where it can move between postsynaptic and presynaptic neurons. It inhibits neurotransmitter release by acting as a zinc endopeptidase that cleaves the '60-Lys-|-Leu-61' bond of synaptobrevins-1 and -2.
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 By similarity.
Subunit structure	Disulfide-linked heterodimer of a light chain (L) and a heavy chain (H). The light chain has the pharmacological activity, while the N- and C-terminal of the heavy chain mediate channel formation and toxin binding, respectively.
Subcellular location	Botulinum neurotoxin D light chain: Secreted. Host cytoplasm › host cytosol. Botulinum neurotoxin D heavy chain: Secreted. Host cell junction › host synapse › host presynaptic cell membrane Potential.
Miscellaneous	There are seven antigenically distinct forms of botulinum neurotoxin: Types A, B, C1, D, E, F, and G. Botulinum type D neurotoxin is synthesized by D strains of C.botulinum which carry the appropriate bacteriophage.
Sequence similarities	Belongs to the peptidase M27 family.

Ontologies

Keywords
Cellular component	Host cell junction Host cell membrane Host cytoplasm Host membrane Host synapse Membrane Secreted
Domain	Transmembrane
Ligand	Metal-binding Zinc
Molecular function	Hydrolase Metalloprotease Neurotoxin Protease Toxin
PTM	Disulfide bond
Technical term	3D-structure Direct protein sequencing
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: InterPro
Cellular component	cell junction Inferred from electronic annotation. Source: UniProtKB-KW cytosol Inferred from Experiment. Source: Reactome endocytic vesicle membrane Inferred from Experiment. Source: Reactome host cell cytosol Inferred from electronic annotation. Source: UniProtKB-SubCell integral to membrane Inferred from electronic annotation. Source: UniProtKB-KW synapse Inferred from electronic annotation. Source: UniProtKB-KW
Molecular function	metalloendopeptidase activity Inferred from electronic annotation. Source: InterPro toxin receptor binding Inferred from electronic annotation. Source: InterPro zinc ion binding Inferred from electronic annotation. Source: UniProtKB-KW
Complete GO annotation...

Sequence annotation (Features)

Feature key

Position(s)

Length

Description

Graphical view

Feature identifier

Molecule processing

Chain

1 – 442

442

Botulinum neurotoxin D light chain

PRO_0000029219

Chain

443 – 1276

834

Botulinum neurotoxin D heavy chain

PRO_0000029220

Sites

Active site

230

1

By similarity

Metal binding

229

1

Zinc; catalytic

Metal binding

233

1

Zinc; catalytic

Metal binding

269

1

Zinc; catalytic

Amino acid modifications

Disulfide bond

437 ↔ 450

Interchain (between light and heavy chains) Probable

Natural variations

Natural variant

15 – 16

2

ND → PV in strain: D-SA.

Natural variant

17 – 18

2

ND → LQ in strain: D-1873.

Natural variant

452

1

K → Q in strain: D-SA.

Natural variant

457

1

R → F in strain: D-1873.

Natural variant

457

1

R → T in strain: D-SA.

Natural variant

462

1

A → D in strain: D-1873.

Natural variant

489

1

K → N in strain: CB16.

Natural variant

644

1

N → K in strain: CB16.

Natural variant

1122

1

Q → R in strain: CB16.

Secondary structure

1

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1276

Helix Strand Turn

Details...

Beta strand

16 – 23

8

Beta strand

34 – 40

7

Beta strand

43 – 46

4

Turn

67 – 69

3

Turn

74 – 77

4

Helix

80 – 97

18

Helix

101 – 112

12

Beta strand

126 – 128

3

Turn

132 – 134

3

Beta strand

136 – 142

7

Beta strand

145 – 152

8

Beta strand

155 – 159

5

Helix

181 – 183

3

Beta strand

184 – 186

3

Beta strand

190 – 193

4

Beta strand

201 – 203

3

Beta strand

218 – 220

3

Helix

223 – 238

16

Helix

267 – 273

7

Helix

275 – 280

6

Helix

283 – 306

24

Beta strand

309 – 312

4

Helix

313 – 318

6

Helix

319 – 329

11

Helix

344 – 355

12

Helix

360 – 366

7

Turn

390 – 392

3

Turn

395 – 397

3

Helix

403 – 405

3

Helix

411 – 413

3

Turn

415 – 417

3

Beta strand

421 – 423

3

Sequences

	Sequence		Length	Mass (Da)
	P19321-1 [UniParc]. Last modified November 1, 1990. Version 1. Checksum: C1EC50F46C8233E2	FASTA	1,276	146,872
	10 20 30 40 50 60 MTWPVKDFNY SDPVNDNDIL YLRIPQNKLI TTPVKAFMIT QNIWVIPERF SSDTNPSLSK 70 80 90 100 110 120 PPRPTSKYQS YYDPSYLSTD EQKDTFLKGI IKLFKRINER DIGKKLINYL VVGSPFMGDS 130 140 150 160 170 180 STPEDTFDFT RHTTNIAVEK FENGSWKVTN IITPSVLIFG PLPNILDYTA SLTLQGQQSN 190 200 210 220 230 240 PSFEGFGTLS ILKVAPEFLL TFSDVTSNQS SAVLGKSIFC MDPVIALMHE LTHSLHQLYG 250 260 270 280 290 300 INIPSDKRIR PQVSEGFFSQ DGPNVQFEEL YTFGGLDVEI IPQIERSQLR EKALGHYKDI 310 320 330 340 350 360 AKRLNNINKT IPSSWISNID KYKKIFSEKY NFDKDNTGNF VVNIDKFNSL YSDLTNVMSE 370 380 390 400 410 420 VVYSSQYNVK NRTHYFSRHY LPVFANILDD NIYTIRDGFN LTNKGFNIEN SGQNIERNPA 430 440 450 460 470 480 LQKLSSESVV DLFTKVCLRL TKNSRDDSTC IKVKNNRLPY VADKDSISQE IFENKIITDE 490 500 510 520 530 540 TNVQNYSDKF SLDESILDGQ VPINPEIVDP LLPNVNMEPL NLPGEEIVFY DDITKYVDYL 550 560 570 580 590 600 NSYYYLESQK LSNNVENITL TTSVEEALGY SNKIYTFLPS LAEKVNKGVQ AGLFLNWANE 610 620 630 640 650 660 VVEDFTTNIM KKDTLDKISD VSVIIPYIGP ALNIGNSALR GNFNQAFATA GVAFLLEGFP 670 680 690 700 710 720 EFTIPALGVF TFYSSIQERE KIIKTIENCL EQRVKRWKDS YQWMVSNWLS RITTQFNHIN 730 740 750 760 770 780 YQMYDSLSYQ ADAIKAKIDL EYKKYSGSDK ENIKSQVENL KNSLDVKISE AMNNINKFIR 790 800 810 820 830 840 ECSVTYLFKN MLPKVIDELN KFDLRTKTEL INLIDSHNII LVGEVDRLKA KVNESFENTM 850 860 870 880 890 900 PFNIFSYTNN SLLKDIINEY FNSINDSKIL SLQNKKNALV DTSGYNAEVR VGDNVQLNTI 910 920 930 940 950 960 YTNDFKLSSS GDKIIVNLNN NILYSAIYEN SSVSFWIKIS KDLTNSHNEY TIINSIEQNS 970 980 990 1000 1010 1020 GWKLCIRNGN IEWILQDVNR KYKSLIFDYS ESLSHTGYTN KWFFVTITNN IMGYMKLYIN 1030 1040 1050 1060 1070 1080 GELKQSQKIE DLDEVKLDKT IVFGIDENID ENQMLWIRDF NIFSKELSNE DINIVYEGQI 1090 1100 1110 1120 1130 1140 LRNVIKDYWG NPLKFDTEYY IINDNYIDRY IAPESNVLVL VQYPDRSKLY TGNPITIKSV 1150 1160 1170 1180 1190 1200 SDKNPYSRIL NGDNIILHML YNSRKYMIIR DTDTIYATQG GECSQNCVYA LKLQSNLGNY 1210 1220 1230 1240 1250 1260 GIGIFSIKNI VSKNKYCSQI FSSFRENTML LADIYKPWRF SFKNAYTPVA VTNYETKLLS 1270 TSSFWKFISR DPGWVE

References

[1]	"Nucleotide sequence of the gene encoding Clostridium botulinum neurotoxin type D." Binz T., Kurazono H., Popoff M.R., Eklund M.W., Sakaguchi G., Kozaki S., Krieglstein K., Henschen A., Gill D.M., Niemann H. Nucleic Acids Res. 18:5556-5556(1990) [PubMed: 2216736] [Abstract] Cited for: NUCLEOTIDE SEQUENCE [GENOMIC DNA]. Strain: Type D / BVD/-3.
[2]	"The complete amino acid sequence of the Clostridium botulinum type D neurotoxin, deduced by nucleotide sequence analysis of the encoding phage d-16 phi genome." Sunagawa H., Ohyama T., Watanabe T., Inoue K. J. Vet. Med. Sci. 54:905-913(1992) [PubMed: 1420572] [Abstract] Cited for: NUCLEOTIDE SEQUENCE [GENOMIC DNA]. Strain: Type D / CB-16.
[3]	"Molecular diversity of neurotoxins from Clostridium botulinum type D strains." Moriishi K., Syuto B., Kubo S., Oguma K. Infect. Immun. 57:2886-2891(1989) [PubMed: 2668193] [Abstract] Cited for: PARTIAL PROTEIN SEQUENCE. Strain: Type D / D-1873 and Type D / South African.
[4]	"Cleavage of members of the synaptobrevin/VAMP family by types D and F botulinal neurotoxins and tetanus toxin." Yamasaki S., Baumeister A., Binz T., Blasi J., Link E., Cornille F., Roques B., Fykse E.M., Suedhof T.C., Jahn R., Niemann H. J. Biol. Chem. 269:12764-12772(1994) [PubMed: 8175689] [Abstract] Cited for: IDENTIFICATION OF SUBSTRATE.
[5]	"Structure of botulinum neurotoxin type D light chain at 1.65 A resolution: repercussions for VAMP-2 substrate specificity." Arndt J.W., Chai Q., Christian T., Stevens R.C. Biochemistry 45:3255-3262(2006) [PubMed: 16519520] [Abstract] Cited for: X-RAY CRYSTALLOGRAPHY (1.65 ANGSTROMS) OF 1-436 IN COMPLEX WITH ZINC IONS.

Web resources

BotDB - A Database Resource for Clostridial Neurotoxins

Cross-references

Sequence databases

EMBLGenBankDDBJ

X54254 Genomic DNA. Translation: CAA38175.1.
S49407 Genomic DNA. Translation: AAB24244.1.

PIR

S11455.

3D structure databases

PDBPDBe

Entry	Method	Resolution (Å)	Chain	Positions	PDBsum
2FPQ	X-ray	1.65	A	1-436	[»]

ModBase

Search...

Enzyme and pathway databases

BRENDA

3.4.24.69. 19133.

Pathway_Interaction_DB

botulinumtoxinpathway. Effects of Botulinum toxin.

Reactome

REACT_11184. Botulinum neurotoxicity.

Family and domain databases

InterPro

IPR008985. ConA-like_lec_gl.
IPR013320. ConA-like_subgrp.
IPR011065. Kunitz_inhibitor_ST1-like.
IPR000395. Peptidase_M27.
IPR013104. Toxin_rcpt_bd_C.
IPR012928. Toxin_rcpt_bd_N.
IPR012500. Toxin_trans.
[Graphical view]

Gene3D

G3DSA:2.60.120.200. ConA_like_subgrp. 1 hit.
G3DSA:3.90.1240.10. Peptidase_M27. 1 hit.

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.

PROSITE

PS00142. ZINC_PROTEASE. 1 hit.
[Graphical view]

ProtoNet

Search...

Entry information

Entry name

BXD_CLOBO

Accession

Primary (citable) accession number: P19321

Entry history

Integrated into UniProtKB/Swiss-Prot:	November 1, 1990
Last sequence update:	November 1, 1990
Last modified:	November 24, 2009
This is version 96 of the entry and version 1 of the sequence. [Complete history]

Entry status

Reviewed (UniProtKB/Swiss-Prot)

Annotation project

HAMAP (High-quality Automated and Manual Annotation of microbial Proteomes)