P15358 (ANTA_HAEOF) Reviewed, UniProtKB/Swiss-Prot

Names and origin

Protein names	Recommended name: Antistasin Short name=ATS Alternative name(s): Blood coagulation factor Xa/proclotting enzyme inhibitor
Organism	Haementeria officinalis (Mexican leech)
Taxonomic identifier	6410 [NCBI]
Taxonomic lineage	Eukaryota › Metazoa › Annelida › Clitellata › Hirudinida › Hirudinea › Rhynchobdellida › Glossiphoniidae › Haementeria

Protein attributes

Sequence length	136 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	This highly disulfide-bonded protein is a potent inhibitor of factor Xa. May have therapeutic utility as an anticoagulant. Also exhibits a strong metastatic activity.
Subcellular location	Secreted.
Miscellaneous	Binds to heparin-agarose, binds to sulfated glycoconjugates. At least four isoforms of antistasin have been identified in leech salivary gland extracts, which differ by 1 or 2 amino-acid residues.
Sequence similarities	Belongs to the protease inhibitor I15 (antistasin) family. [View classification] Contains 2 antistasin-like domains.

Ontologies

Keywords
Biological process	Blood coagulation
Cellular component	Secreted
Domain	Repeat Signal
Ligand	Heparin-binding
Molecular function	Protease inhibitor Serine protease inhibitor
PTM	Disulfide bond Pyrrolidone carboxylic acid
Technical term	3D-structure Direct protein sequencing
Gene Ontology (GO)
Biological process	blood coagulation Inferred from electronic annotation. Source: UniProtKB-KW negative regulation of coagulation Inferred from electronic annotation. Source: InterPro
Cellular component	extracellular region Inferred from electronic annotation. Source: UniProtKB-SubCell
Molecular function	heparin binding Inferred from electronic annotation. Source: UniProtKB-KW serine-type endopeptidase inhibitor activity 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

Signal peptide

1 – 17

17

Ref.2 Ref.3

Chain

18 – 136

119

Antistasin

PRO_0000001700

Regions

Domain

45 – 70

26

Antistasin-like 1

Domain

100 – 125

26

Antistasin-like 2

Region

114 – 117

4

Heparin-binding Potential

Region

128 – 135

8

Heparin-binding Potential

Sites

Site

51 – 52

2

Reactive bond

Site

106 – 107

2

Reactive bond

Amino acid modifications

Modified residue

18

1

Pyrrolidone carboxylic acid

Disulfide bond

25 ↔ 36

Disulfide bond

30 ↔ 43

Disulfide bond

45 ↔ 65

Disulfide bond

50 ↔ 68

Disulfide bond

54 ↔ 70

Disulfide bond

79 ↔ 90

Disulfide bond

84 ↔ 97

Disulfide bond

99 ↔ 120

Disulfide bond

105 ↔ 123

Disulfide bond

109 ↔ 125

Natural variations

Natural variant

22

1

G → R in isoform B.

Natural variant

47

1

G → E.

Natural variant

52

1

M → V.

Natural variant

71

1

R → I.

Secondary structure

1

.

.

.

.

.

.

.

.

.

.

.

.

.

.

.

.

.

.

.

.

136

Helix Strand Turn

Details...

Helix

25 – 28

4

Turn

38 – 40

3

Beta strand

58 – 60

3

Beta strand

66 – 70

5

Helix

81 – 83

3

Turn

92 – 94

3

Beta strand

95 – 97

3

Beta strand

99 – 101

3

Beta strand

113 – 115

3

Beta strand

121 – 125

5

Sequences

	Sequence		Length	Mass (Da)
	P15358 [UniParc]. Last modified February 1, 1991. Version 2. Checksum: 582AF009ED9A0291	FASTA	136	15,225
	10 20 30 40 50 60 MIKLAILLLF TVAIVRCQGP FGPGCEEAGC PEGSACNIIT DRCTCSGVRC RMHCPHGFQR 70 80 90 100 110 120 SRYGCEFCKC RLEPMKATCD ISECPEGMMC SRLTNKCDCK IDINCRKTCP NGLKRDKLGC 130 EYCECRPKRK LIPRLS

References

[1]	"Cloning and expression of cDNA encoding antistasin, a leech-derived protein having anti-coagulant and anti-metastatic properties." Han J.H., Law S.W., Keller P.M., Kniskern P.J., Silberklang M., Tung J.S., Gasic T.B., Gasic G.J., Friedman P.A., Ellis R.W. Gene 75:47-57(1989) [PubMed: 2470652] [Abstract] Cited for: NUCLEOTIDE SEQUENCE [MRNA].
[2]	"The amino acid sequence of antistasin. A potent inhibitor of factor Xa reveals a repeated internal structure." Nutt E., Gasic T., Rodkey J., Gasic G.J., Jacobs J.W., Friedman P.A., Simpson E. J. Biol. Chem. 263:10162-10167(1988) [PubMed: 3164720] [Abstract] Cited for: PROTEIN SEQUENCE OF 18-136. Tissue: Saliva.
[3]	"Purification and characterization of inhibitors of blood coagulation factor Xa from hematophagous organisms." Dunwiddie C.T., Waxman L., Vlasuk G.P., Friedman P.A. Methods Enzymol. 223:291-312(1993) [PubMed: 8271959] [Abstract] Cited for: PROTEIN SEQUENCE OF 18-136. Tissue: Saliva.
[4]	"Antistasin, a leech-derived inhibitor of factor Xa. Kinetic analysis of enzyme inhibition and identification of the reactive site." Dunwiddie C., Thornberry N.A., Bull H.G., Sardana M., Friedman P.A., Jacobs J.W., Simpson E. J. Biol. Chem. 264:16694-16699(1989) [PubMed: 2777803] [Abstract] Cited for: REACTIVE SITE.
[5]	"Antistasin, an inhibitor of coagulation and metastasis, binds to sulfatide (Gal(3-SO4) beta 1-1Cer) and has a sequence homology with other proteins that bind sulfated glycoconjugates." Holt G.D., Krivan H.C., Gasic G.J., Ginsburg V. J. Biol. Chem. 264:12138-12140(1989) [PubMed: 2745433] [Abstract] Cited for: SULFATIDE-BINDING.
[6]	"Site-directed mutagenesis of the leech-derived factor Xa inhibitor antistasin. Probing of the reactive site." Hofmann K.J., Nutt E.M., Dunwiddie C. Biochem. J. 287:943-949(1992) [PubMed: 1445252] [Abstract] Cited for: MUTAGENESIS.
[7]	"Mutational analysis of antistasin, an inhibitor of blood coagulation factor Xa derived from the Mexican leech Haementeria officinalis." Theunissen H.J., Dijkema R., Swinkels J.C., de Poorter T.L., Vink P.M., van Dinther T.G. Thromb. Res. 75:41-50(1994) [PubMed: 8073407] [Abstract] Cited for: MUTAGENESIS.
[8]	"X-ray structure of antistasin at 1.9-A resolution and its modelled complex with blood coagulation factor Xa." Lapatto R., Krengel U., Schreuder H.A., Arkema A., de Boer B., Kalk K.H., Hol W.G.J., Grootenhuis P.D.J., Mulders J.W.M., Dijkema R., Theunissen H.J.M., Dijkstra B.W. EMBO J. 16:5151-5161(1997) [PubMed: 9311976] [Abstract] Cited for: X-RAY CRYSTALLOGRAPHY (1.9 ANGSTROMS) OF 24-127.

Cross-references

Sequence databases

EMBL
GenBank
DDBJ

M24422 mRNA. Translation: AAA29192.1.
M24423 mRNA. Translation: AAA29193.1.

PIR

A28806.
A34398.
JS0209.
S13904.

3D structure databases

PDBe
RCSB PDB
PDBj

Entry	Method	Resolution (Å)	Chain	Positions	PDBsum
1SKZ	X-ray	1.90	A	20-136	[»]

ProteinModelPortal

P15358.

SMR

P15358. Positions 24-127.

ModBase

Search...

Protein family/group databases

MEROPS

I15.007.

Protocols and materials databases

StructuralBiologyKnowledgebase

Search...

Family and domain databases

InterPro

IPR011061. Prot_inh_I14/15_hirudin/antisn.
IPR018112. Prot_inh_I15_antistasin.
IPR004094. Prot_inh_I15_antistasin-like.
IPR008086. Prot_inh_I15_antistasin_leech.
[Graphical view]

Gene3D

G3DSA:2.10.22.10. Prot_inh_antistn. 2 hits.

Pfam

PF02822. Antistasin. 1 hit.
[Graphical view]

PRINTS

PR01706. ANTISTASIN.

SUPFAM

SSF57262. Antihaemostatic. 2 hits.

PROSITE

PS51252. ANTISTASIN. 2 hits.
[Graphical view]

ProtoNet

Search...

Entry information

Entry name

ANTA_HAEOF

Accession

Primary (citable) accession number: P15358
Secondary accession number(s): Q9TWQ8, Q9TX45

Entry history

Integrated into UniProtKB/Swiss-Prot:	April 1, 1990
Last sequence update:	February 1, 1991
Last modified:	November 16, 2011
This is version 83 of the entry and version 2 of the sequence. [Complete history]

Entry status

Reviewed (UniProtKB/Swiss-Prot)