Reviewed, UniProtKB/Swiss-Prot P01050 (HIRV1_HIRME)

Names and origin

Protein names	Recommended name:     Hirudin variant-1 Alternative name(s):     Hirudin-1     Hirudin-I     Lepirudin
Organism	Hirudo medicinalis (Medicinal leech)
Taxonomic identifier	6421 [NCBI]
Taxonomic lineage	Eukaryota › Metazoa › Annelida › Clitellata › Hirudinida › Hirudinea › Arhynchobdellida › Hirudiniformes › Hirudinidae › Hirudo

Protein attributes

Sequence length	65 AA.
Sequence status	Complete.
Protein existence	Evidence at protein level.

General annotation (Comments)

Function	Hirudin is a potent thrombin-specific protease inhibitor. It forms a stable non-covalent complex with alpha-thrombin, thereby abolishing its ability to cleave fibrinogen. Ref.13
Subcellular location	Secreted.
Pharmaceutical use	Available under the name Refludan (Hoechst Marion Roussel). Used to treat heparin-induced thrombocytopenia (HIT).
Sequence similarities	Belongs to the protease inhibitor I14 (hirudin) family. [View classification]

Ontologies

Keywords
Cellular component	Secreted
Molecular function	Protease inhibitor Serine protease inhibitor
PTM	Disulfide bond Glycoprotein Sulfation
Technical term	3D-structure Direct protein sequencing Pharmaceutical
Gene Ontology (GO)
Cellular component	extracellular region Inferred from electronic annotation. Source: UniProtKB-SubCell
Molecular function	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

Chain

1 – 65

65

Hirudin variant-1

PRO_0000195639

Regions

Region

1 – 3

3

Interaction with thrombin active site

Region

55 – 65

11

Interaction with fibrinogen-binding exosite of thrombin

Amino acid modifications

Modified residue

63

1

Sulfotyrosine Ref.12 Ref.14

Glycosylation

45

1

O-linked (GalNAc...) By similarity

Disulfide bond

6 ↔ 14

Ref.13 Ref.14 Ref.7

Disulfide bond

16 ↔ 28

Ref.13 Ref.14 Ref.7

Disulfide bond

22 ↔ 39

Ref.13 Ref.14 Ref.7

Experimental info

Mutagenesis

1

1

V → E or K: Strongly decreased affinity for thrombin. Ref.5

Mutagenesis

1

1

V → G or S: Decreased affinity for thrombin. Ref.5

Mutagenesis

1

1

V → L or R: No effect. Ref.5

Mutagenesis

2

1

V → E or G: Strongly decreased affinity for thrombin. Ref.5

Mutagenesis

2

1

V → L: Decreased affinity for thrombin. Ref.5

Mutagenesis

3

1

Y → A: Strongly decreased affinity for thrombin. Ref.5

Mutagenesis

4

1

T → A: Decreased affinity for thrombin. Ref.5

Mutagenesis

5

1

D → A or E: Decreased affinity for thrombin. Ref.5

Mutagenesis

15

1

L → A: Strongly decreased affinity for thrombin. Ref.6

Mutagenesis

17

1

E → A: Slightly decreased affinity for thrombin.

Mutagenesis

20

1

N → A: Decreased affinity for thrombin. Ref.6

Mutagenesis

21

1

V → A: Slightly decreased affinity for thrombin. Ref.6

Mutagenesis

56

1

F → A or W: Slightly decreased affinity for thrombin. Ref.4

Mutagenesis

56

1

F → I, L, T or V: Strongly decreased affinity for thrombin. Ref.4

Mutagenesis

56

1

F → Y: No effect. Ref.4

Mutagenesis

60

1

P → A or G: Decreased affinity for thrombin. Ref.4

Mutagenesis

63

1

Y → A, E or L: Slightly decreased affinity for thrombin. Ref.4

Mutagenesis

63

1

Y → F: No effect. Ref.4

Mutagenesis

63

1

Y → V: Decreased affinity for thrombin. Ref.4

Secondary structure

1

.

.

.

.

.

.

.

.

.

.

.

65

Helix Strand Turn

Details...

Beta strand

13 – 15

3

Beta strand

17 – 19

3

Beta strand

26 – 29

4

Beta strand

38 – 42

5

Helix

61 – 63

3

Sequences

	Sequence		Length	Mass (Da)
	P01050-1 [UniParc]. Last modified July 21, 1986. Version 1. Checksum: 9085A5876E3DE9FF	FASTA	65	6,970
	10 20 30 40 50 60 VVYTDCTESG QNLCLCEGSN VCGQGNKCIL GSDGEKNQCV TGEGTPKPQS HNDGDFEEIP EEYLQ

References

[1]	"The complete amino acid sequence of hirudin, a thrombin specific inhibitor. Application of colour carboxymethylation." Dodt J., Mueller H.-P., Seemueller U., Chang J.-Y. FEBS Lett. 165:180-183(1984) Cited for: PROTEIN SEQUENCE.
[2]	Petersen T.E., Roberts H.R., Sottrup-Jensen L., Magnusson S., Bagdy D. (In) Peeters H. (eds.); Protides of the biological fluids, Proc. 23th colloquium, pp.145-149, Pergamon Press, New York (1976) Cited for: PROTEIN SEQUENCE.
[3]	"Primary structures of new 'iso-hirudins'." Scharf M., Engels J., Tripier D. FEBS Lett. 255:105-110(1989) [PubMed: 2792365] [Abstract] Cited for: PROTEIN SEQUENCE OF 1-17 AND 28-65.
[4]	"Role of interactions involving C-terminal nonpolar residues of hirudin in the formation of the thrombin-hirudin complex." Betz A., Hofsteenge J., Stone S.R. Biochemistry 30:9848-9853(1991) [PubMed: 1911777] [Abstract] Cited for: MUTAGENESIS OF PHE-56; PRO-60 AND TYR-63.
[5]	"Interaction of the N-terminal region of hirudin with the active-site cleft of thrombin." Betz A., Hofsteenge J., Stone S.R. Biochemistry 31:4557-4562(1992) [PubMed: 1581311] [Abstract] Cited for: MUTAGENESIS OF VAL-1; VAL-2; TYR-3; THR-4 AND ASP-5.
[6]	"Contribution of interactions with the core domain of hirudin to the stability of its complex with thrombin." Betz A., Hopkins P.C.R., Le Bonniec B.F., Stone S.R. Biochem. J. 298:507-510(1994) [PubMed: 8135762] [Abstract] Cited for: MUTAGENESIS OF LEU-15; ASN-20 AND VAL-21.
[7]	"Solution structure of recombinant hirudin and the Lys-47-->Glu mutant: a nuclear magnetic resonance and hybrid distance geometry-dynamical simulated annealing study." Folkers P.J.M., Clore G.M., Driscoll P.C., Dodt J., Koehler S., Gronenborn A.M. Biochemistry 28:2601-2617(1989) [PubMed: 2567183] [Abstract] Cited for: STRUCTURE BY NMR, DISULFIDE BONDS.
[8]	"Conformation of recombinant desulfatohirudin in aqueous solution determined by nuclear magnetic resonance." Haruyama H., Wuethrich K. Biochemistry 28:4301-4312(1989) [PubMed: 2765488] [Abstract] Cited for: STRUCTURE BY NMR.
[9]	"Crystal structure of the thrombin-hirudin complex: a novel mode of serine protease inhibition." Gruetter M.G., Priestle J.P., Rahuel J., Grossenbacher H., Bode W., Hofsteenge J., Stone S.R. EMBO J. 9:2361-2365(1990) [PubMed: 2369893] [Abstract] Cited for: X-RAY CRYSTALLOGRAPHY (2.95 ANGSTROMS) IN COMPLEX WITH THROMBIN.
[10]	"The structure of a complex of bovine alpha-thrombin and recombinant hirudin at 2.8-A resolution." Vitali J., Martin P.D., Malkowski M.G., Robertson W.D., Lazar J.B., Winant R.C., Johnson P.H., Edwards B.F.P. J. Biol. Chem. 267:17670-17678(1992) [PubMed: 1517214] [Abstract] Cited for: X-RAY CRYSTALLOGRAPHY (2.8 ANGSTROMS) OF 51-65 IN COMPLEX WITH THROMBIN.
[11]	"Nuclear magnetic resonance solution structure of hirudin(1-51) and comparison with corresponding three-dimensional structures determined using the complete 65-residue hirudin polypeptide chain." Szyperski T., Guentert P., Stone S.R., Wuethrich K. J. Mol. Biol. 228:1193-1205(1992) [PubMed: 1335515] [Abstract] Cited for: STRUCTURE BY NMR OF 1-51.
[12]	"Changes in interactions in complexes of hirudin derivatives and human alpha-thrombin due to different crystal forms." Priestle J.P., Rahuel J., Rink H., Tones M., Gruetter M.G. Protein Sci. 2:1630-1642(1993) [PubMed: 8251938] [Abstract] Cited for: X-RAY CRYSTALLOGRAPHY (2.2 ANGSTROMS) OF 54-65 IN COMPLEX WITH THROMBIN, SULFATION AT TYR-63.
[13]	"The NMR solution structure of recombinant RGD-hirudin." Song X., Mo W., Liu X., Zhu L., Yan X., Song H., Dai L. Biochem. Biophys. Res. Commun. 360:103-108(2007) [PubMed: 17585879] [Abstract] Cited for: STRUCTURE BY NMR OF 1-57 OF MUTANT 32-R--D-34, FUNCTION, DISULFIDE BONDS.
[14]	"Crystal structure of a biosynthetic sulfo-hirudin complexed to thrombin." Liu C.C., Brustad E., Liu W., Schultz P.G. J. Am. Chem. Soc. 129:10648-10649(2007) [PubMed: 17685615] [Abstract] Cited for: X-RAY CRYSTALLOGRAPHY (1.84 ANGSTROMS), DISULFIDE BONDS, SULFATION AT TYR-63.
+	Additional computationally mapped references.

Web resources

Refludan

Clinical information on Refludan

Cross-references

Sequence databases

PIR

HULXH. A91318.
S05672.

3D structure databases

PDBe
RCSB PDB
PDBj

Entry	Method	Resolution (Å)	Chain	Positions	PDBsum
1AD8	X-ray	2.00	I	55-64	[»]
1AE8	X-ray	2.00	I	55-64	[»]
1AFE	X-ray	2.00	I	55-64	[»]
1AI8	X-ray	1.85	I	55-64	[»]
1AWF	X-ray	2.20	I	55-64	[»]
1BCU	X-ray	2.00	I	54-65	[»]
1D3D	X-ray	2.04	H	54-65	[»]
1D3P	X-ray	2.10	H	54-65	[»]
1D3Q	X-ray	2.90	H	54-65	[»]
1D3T	X-ray	3.00	H	54-65	[»]
1D4P	X-ray	2.07	H	54-65	[»]
1H8D	X-ray	1.40	I	55-64	[»]
1H8I	X-ray	1.75	I	55-64	[»]
1HIC	NMR	-	A	1-51	[»]
1HRT	X-ray	2.80	I	1-65	[»]
1HXE	X-ray	2.10	I	55-64	[»]
1HXF	X-ray	2.10	I	55-64	[»]
1QHR	X-ray	2.20	I	55-64	[»]
1QJ1	X-ray	2.00	I	55-64	[»]
1QJ6	X-ray	2.20	I	55-64	[»]
1QJ7	X-ray	2.20	I	55-64	[»]
1TMT	X-ray	2.20	I	53-65	[»]
1TMU	X-ray	2.50	I	55-65	[»]
1UMA	X-ray	2.00	I	55-64	[»]
1WBG	X-ray	2.20	I	55-64	[»]
2HIR	NMR	-	A	1-65	[»]
2JOO	NMR	-	A	1-61	[»]
2PW8	X-ray	1.84	I	3-65	[»]
2UUF	X-ray	1.26	H	55-64	[»]
2UUJ	X-ray	1.32	H	55-64	[»]
2UUK	X-ray	1.39	H	55-64	[»]
2V3O	X-ray	1.79	I	56-65	[»]
2ZC9	X-ray	1.58	I	54-64	[»]
2ZDA	X-ray	1.73	I	54-64	[»]
2ZDV	X-ray	1.72	I	54-64	[»]
2ZF0	X-ray	2.20	I	54-64	[»]
2ZFF	X-ray	1.47	I	54-64	[»]
2ZFP	X-ray	2.25	I	54-64	[»]
2ZGB	X-ray	1.60	I	54-64	[»]
2ZGX	X-ray	1.80	I	54-64	[»]
2ZHQ	X-ray	1.96	I	54-64	[»]
2ZI2	X-ray	1.65	I	54-64	[»]
2ZIQ	X-ray	1.65	I	54-64	[»]
2ZNK	X-ray	1.80	I	54-64	[»]
2ZO3	X-ray	1.70	I	54-64	[»]
3D49	X-ray	1.50	I	54-64	[»]
3DT0	X-ray	2.40	I	54-64	[»]
3DUX	X-ray	1.60	I	54-64	[»]
3EQ0	X-ray	1.53	I	54-64	[»]
4HIR	NMR	-	A	1-65	[»]
5HIR	NMR	-	A	1-65	[»]
6HIR	NMR	-	A	1-65	[»]

DisProt

DP00137.

ModBase

Search...

Protein family/group databases

MEROPS

I14.001.

Family and domain databases

InterPro

IPR000429. Prot_inh_hirudin.
IPR011061. Prot_inh_I14/15_hirudin/antisn.
[Graphical view]

Gene3D

G3DSA:2.70.10.10. Prot_inh_hirudin. 1 hit.

Pfam

PF00713. Hirudin. 1 hit.
[Graphical view]

PIRSF

PIRSF001640. Hirudin. 1 hit.

PRINTS

PR00777. HIRUDIN.

ProDom

PD004216. Prot_inh_hirudin. 1 hit.
[Graphical view] [Entries sharing at least one domain]

ProtoNet

Search...

Other Resources

DrugBank

DB00001. Lepirudin.

Entry information

Entry name

HIRV1_HIRME

Accession

Primary (citable) accession number: P01050
Secondary accession number(s): P28501

Entry history

Integrated into UniProtKB/Swiss-Prot:	July 21, 1986
Last sequence update:	July 21, 1986
Last modified:	December 15, 2009
This is version 85 of the entry and version 1 of the sequence. [Complete history]

Entry status

Reviewed (UniProtKB/Swiss-Prot)