HIRV1_HIRME
ID HIRV1_HIRME Reviewed; 65 AA.
AC P01050; P28501;
DT 21-JUL-1986, integrated into UniProtKB/Swiss-Prot.
DT 21-JUL-1986, sequence version 1.
DT 25-MAY-2022, entry version 134.
DE RecName: Full=Hirudin variant-1;
DE AltName: Full=Hirudin-1;
DE AltName: Full=Hirudin-I;
DE AltName: INN=Lepirudin;
OS Hirudo medicinalis (Medicinal leech).
OC Eukaryota; Metazoa; Spiralia; Lophotrochozoa; Annelida; Clitellata;
OC Hirudinea; Hirudinida; Hirudiniformes; Hirudinidae; Hirudo.
OX NCBI_TaxID=6421;
RN [1]
RP PROTEIN SEQUENCE.
RA Dodt J., Mueller H.-P., Seemueller U., Chang J.-Y.;
RT "The complete amino acid sequence of hirudin, a thrombin specific
RT inhibitor. Application of colour carboxymethylation.";
RL FEBS Lett. 165:180-183(1984).
RN [2]
RP PROTEIN SEQUENCE.
RA Petersen T.E., Roberts H.R., Sottrup-Jensen L., Magnusson S., Bagdy D.;
RL (In) Peeters H. (eds.);
RL Protides of the biological fluids, Proc. 23th colloquium, pp.145-149,
RL Pergamon Press, New York (1976).
RN [3]
RP PROTEIN SEQUENCE OF 1-17 AND 28-65.
RX PubMed=2792365; DOI=10.1016/0014-5793(89)81070-1;
RA Scharf M., Engels J., Tripier D.;
RT "Primary structures of new 'iso-hirudins'.";
RL FEBS Lett. 255:105-110(1989).
RN [4]
RP MUTAGENESIS OF PHE-56; PRO-60 AND TYR-63.
RX PubMed=1911777; DOI=10.1021/bi00105a006;
RA Betz A., Hofsteenge J., Stone S.R.;
RT "Role of interactions involving C-terminal nonpolar residues of hirudin in
RT the formation of the thrombin-hirudin complex.";
RL Biochemistry 30:9848-9853(1991).
RN [5]
RP MUTAGENESIS OF VAL-1; VAL-2; TYR-3; THR-4 AND ASP-5.
RX PubMed=1581311; DOI=10.1021/bi00134a004;
RA Betz A., Hofsteenge J., Stone S.R.;
RT "Interaction of the N-terminal region of hirudin with the active-site cleft
RT of thrombin.";
RL Biochemistry 31:4557-4562(1992).
RN [6]
RP MUTAGENESIS OF LEU-15; ASN-20 AND VAL-21.
RX PubMed=8135762; DOI=10.1042/bj2980507;
RA Betz A., Hopkins P.C.R., Le Bonniec B.F., Stone S.R.;
RT "Contribution of interactions with the core domain of hirudin to the
RT stability of its complex with thrombin.";
RL Biochem. J. 298:507-510(1994).
RN [7]
RP STRUCTURE BY NMR, AND DISULFIDE BONDS.
RX PubMed=2567183; DOI=10.1021/bi00432a038;
RA Folkers P.J.M., Clore G.M., Driscoll P.C., Dodt J., Koehler S.,
RA Gronenborn A.M.;
RT "Solution structure of recombinant hirudin and the Lys-47-->Glu mutant: a
RT nuclear magnetic resonance and hybrid distance geometry-dynamical simulated
RT annealing study.";
RL Biochemistry 28:2601-2617(1989).
RN [8]
RP STRUCTURE BY NMR.
RX PubMed=2765488; DOI=10.1021/bi00436a027;
RA Haruyama H., Wuethrich K.;
RT "Conformation of recombinant desulfatohirudin in aqueous solution
RT determined by nuclear magnetic resonance.";
RL Biochemistry 28:4301-4312(1989).
RN [9]
RP X-RAY CRYSTALLOGRAPHY (2.95 ANGSTROMS) IN COMPLEX WITH THROMBIN.
RX PubMed=2369893; DOI=10.1002/j.1460-2075.1990.tb07410.x;
RA Gruetter M.G., Priestle J.P., Rahuel J., Grossenbacher H., Bode W.,
RA Hofsteenge J., Stone S.R.;
RT "Crystal structure of the thrombin-hirudin complex: a novel mode of serine
RT protease inhibition.";
RL EMBO J. 9:2361-2365(1990).
RN [10]
RP X-RAY CRYSTALLOGRAPHY (2.8 ANGSTROMS) OF 51-65 IN COMPLEX WITH THROMBIN.
RX PubMed=1517214; DOI=10.1016/s0021-9258(19)37095-4;
RA Vitali J., Martin P.D., Malkowski M.G., Robertson W.D., Lazar J.B.,
RA Winant R.C., Johnson P.H., Edwards B.F.P.;
RT "The structure of a complex of bovine alpha-thrombin and recombinant
RT hirudin at 2.8-A resolution.";
RL J. Biol. Chem. 267:17670-17678(1992).
RN [11]
RP STRUCTURE BY NMR OF 1-51.
RX PubMed=1335515; DOI=10.1016/0022-2836(92)90325-e;
RA Szyperski T., Guentert P., Stone S.R., Wuethrich K.;
RT "Nuclear magnetic resonance solution structure of hirudin(1-51) and
RT comparison with corresponding three-dimensional structures determined using
RT the complete 65-residue hirudin polypeptide chain.";
RL J. Mol. Biol. 228:1193-1205(1992).
RN [12]
RP X-RAY CRYSTALLOGRAPHY (2.2 ANGSTROMS) OF 54-65 IN COMPLEX WITH THROMBIN,
RP AND SULFATION AT TYR-63.
RX PubMed=8251938; DOI=10.1002/pro.5560021009;
RA Priestle J.P., Rahuel J., Rink H., Tones M., Gruetter M.G.;
RT "Changes in interactions in complexes of hirudin derivatives and human
RT alpha-thrombin due to different crystal forms.";
RL Protein Sci. 2:1630-1642(1993).
RN [13]
RP STRUCTURE BY NMR OF 1-57 OF MUTANT 32-R--D-34, FUNCTION, AND DISULFIDE
RP BONDS.
RX PubMed=17585879; DOI=10.1016/j.bbrc.2007.06.014;
RA Song X., Mo W., Liu X., Zhu L., Yan X., Song H., Dai L.;
RT "The NMR solution structure of recombinant RGD-hirudin.";
RL Biochem. Biophys. Res. Commun. 360:103-108(2007).
RN [14]
RP X-RAY CRYSTALLOGRAPHY (1.84 ANGSTROMS), DISULFIDE BONDS, AND SULFATION AT
RP TYR-63.
RX PubMed=17685615; DOI=10.1021/ja0735002;
RA Liu C.C., Brustad E., Liu W., Schultz P.G.;
RT "Crystal structure of a biosynthetic sulfo-hirudin complexed to thrombin.";
RL J. Am. Chem. Soc. 129:10648-10649(2007).
CC -!- FUNCTION: Hirudin is a potent thrombin-specific protease inhibitor. It
CC forms a stable non-covalent complex with alpha-thrombin, thereby
CC abolishing its ability to cleave fibrinogen.
CC {ECO:0000269|PubMed:17585879}.
CC -!- SUBCELLULAR LOCATION: Secreted.
CC -!- PHARMACEUTICAL: Available under the name Refludan (Hoechst Marion
CC Roussel). Used to treat heparin-induced thrombocytopenia (HIT).
CC -!- SIMILARITY: Belongs to the protease inhibitor I14 (hirudin) family.
CC {ECO:0000305}.
CC -!- WEB RESOURCE: Name=Refludan; Note=Clinical information on Refludan;
CC URL="https://www.rxlist.com/refludan-drug.htm";
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DR PIR; A91318; HULXH.
DR PIR; S05672; S05672.
DR PDB; 1AD8; X-ray; 2.00 A; I=55-64.
DR PDB; 1AE8; X-ray; 2.00 A; I=55-64.
DR PDB; 1AFE; X-ray; 2.00 A; I=55-64.
DR PDB; 1AHT; X-ray; 1.60 A; I=55-64.
DR PDB; 1AI8; X-ray; 1.85 A; I=55-64.
DR PDB; 1AWF; X-ray; 2.20 A; I=55-64.
DR PDB; 1AY6; X-ray; 1.80 A; I=55-64.
DR PDB; 1BA8; X-ray; 1.80 A; C=53-64.
DR PDB; 1BB0; X-ray; 2.10 A; C=53-64.
DR PDB; 1BCU; X-ray; 2.00 A; I=54-65.
DR PDB; 1CA8; X-ray; 2.10 A; C=53-64.
DR PDB; 1D3D; X-ray; 2.04 A; H=54-65.
DR PDB; 1D3P; X-ray; 2.10 A; H=54-65.
DR PDB; 1D3Q; X-ray; 2.90 A; H=54-65.
DR PDB; 1D3T; X-ray; 3.00 A; H=54-65.
DR PDB; 1D4P; X-ray; 2.07 A; H=54-65.
DR PDB; 1H8D; X-ray; 1.40 A; I=55-64.
DR PDB; 1H8I; X-ray; 1.75 A; I=55-64.
DR PDB; 1HIC; NMR; -; A=1-51.
DR PDB; 1HRT; X-ray; 2.80 A; I=1-65.
DR PDB; 1HXE; X-ray; 2.10 A; I=55-64.
DR PDB; 1HXF; X-ray; 2.10 A; I=55-64.
DR PDB; 1NO9; X-ray; 1.90 A; I=55-65.
DR PDB; 1QHR; X-ray; 2.20 A; I=55-64.
DR PDB; 1QJ1; X-ray; 2.00 A; I=55-64.
DR PDB; 1QJ6; X-ray; 2.20 A; I=55-64.
DR PDB; 1QJ7; X-ray; 2.20 A; I=55-64.
DR PDB; 1TMT; X-ray; 2.20 A; I=53-65.
DR PDB; 1TMU; X-ray; 2.50 A; I=55-65.
DR PDB; 1UMA; X-ray; 2.00 A; I=55-64.
DR PDB; 1WBG; X-ray; 2.20 A; I=55-64.
DR PDB; 2HIR; NMR; -; A=1-65.
DR PDB; 2JOO; NMR; -; A=1-65.
DR PDB; 2PW8; X-ray; 1.84 A; I=3-65.
DR PDB; 2UUF; X-ray; 1.26 A; H=55-64.
DR PDB; 2UUJ; X-ray; 1.32 A; H=55-64.
DR PDB; 2UUK; X-ray; 1.39 A; H=55-64.
DR PDB; 2V3O; X-ray; 1.79 A; I=56-65.
DR PDB; 2ZC9; X-ray; 1.58 A; I=54-64.
DR PDB; 2ZDA; X-ray; 1.73 A; I=54-64.
DR PDB; 2ZDV; X-ray; 1.72 A; I=54-64.
DR PDB; 2ZF0; X-ray; 2.20 A; I=54-64.
DR PDB; 2ZFF; X-ray; 1.47 A; I=54-64.
DR PDB; 2ZFP; X-ray; 2.25 A; I=54-64.
DR PDB; 2ZGB; X-ray; 1.60 A; I=54-64.
DR PDB; 2ZGX; X-ray; 1.80 A; I=54-64.
DR PDB; 2ZHQ; X-ray; 1.96 A; I=54-64.
DR PDB; 2ZI2; X-ray; 1.65 A; I=54-64.
DR PDB; 2ZIQ; X-ray; 1.65 A; I=54-64.
DR PDB; 2ZNK; X-ray; 1.80 A; I=54-64.
DR PDB; 2ZO3; X-ray; 1.70 A; I=54-64.
DR PDB; 3D49; X-ray; 1.50 A; I=54-64.
DR PDB; 3DHK; X-ray; 1.73 A; I=54-64.
DR PDB; 3DT0; X-ray; 2.40 A; I=54-64.
DR PDB; 3DUX; X-ray; 1.60 A; I=54-64.
DR PDB; 3EGK; X-ray; 2.20 A; I=54-64.
DR PDB; 3EQ0; X-ray; 1.53 A; I=54-64.
DR PDB; 3LDX; X-ray; 2.25 A; I=55-65.
DR PDB; 3UTU; X-ray; 1.55 A; I=54-64.
DR PDB; 4AX9; X-ray; 1.90 A; I=55-65.
DR PDB; 4BAH; X-ray; 1.94 A; D=53-64.
DR PDB; 4BAK; X-ray; 1.94 A; D=53-64.
DR PDB; 4BAM; X-ray; 1.88 A; D=53-64.
DR PDB; 4BAN; X-ray; 1.87 A; D=53-64.
DR PDB; 4BAQ; X-ray; 1.89 A; D=53-64.
DR PDB; 4HIR; NMR; -; A=1-65.
DR PDB; 4LXB; X-ray; 1.61 A; I=53-65.
DR PDB; 4MLF; X-ray; 2.20 A; D=1-65.
DR PDB; 4YES; X-ray; 1.50 A; H=54-65.
DR PDB; 5HIR; NMR; -; A=1-65.
DR PDB; 6HIR; NMR; -; A=1-65.
DR PDB; 7A0D; X-ray; 1.60 A; III=1-65.
DR PDB; 7A0E; X-ray; 1.90 A; III=1-65.
DR PDB; 7A0F; X-ray; 2.70 A; III=1-65.
DR PDBsum; 1AD8; -.
DR PDBsum; 1AE8; -.
DR PDBsum; 1AFE; -.
DR PDBsum; 1AHT; -.
DR PDBsum; 1AI8; -.
DR PDBsum; 1AWF; -.
DR PDBsum; 1AY6; -.
DR PDBsum; 1BA8; -.
DR PDBsum; 1BB0; -.
DR PDBsum; 1BCU; -.
DR PDBsum; 1CA8; -.
DR PDBsum; 1D3D; -.
DR PDBsum; 1D3P; -.
DR PDBsum; 1D3Q; -.
DR PDBsum; 1D3T; -.
DR PDBsum; 1D4P; -.
DR PDBsum; 1H8D; -.
DR PDBsum; 1H8I; -.
DR PDBsum; 1HIC; -.
DR PDBsum; 1HRT; -.
DR PDBsum; 1HXE; -.
DR PDBsum; 1HXF; -.
DR PDBsum; 1NO9; -.
DR PDBsum; 1QHR; -.
DR PDBsum; 1QJ1; -.
DR PDBsum; 1QJ6; -.
DR PDBsum; 1QJ7; -.
DR PDBsum; 1TMT; -.
DR PDBsum; 1TMU; -.
DR PDBsum; 1UMA; -.
DR PDBsum; 1WBG; -.
DR PDBsum; 2HIR; -.
DR PDBsum; 2JOO; -.
DR PDBsum; 2PW8; -.
DR PDBsum; 2UUF; -.
DR PDBsum; 2UUJ; -.
DR PDBsum; 2UUK; -.
DR PDBsum; 2V3O; -.
DR PDBsum; 2ZC9; -.
DR PDBsum; 2ZDA; -.
DR PDBsum; 2ZDV; -.
DR PDBsum; 2ZF0; -.
DR PDBsum; 2ZFF; -.
DR PDBsum; 2ZFP; -.
DR PDBsum; 2ZGB; -.
DR PDBsum; 2ZGX; -.
DR PDBsum; 2ZHQ; -.
DR PDBsum; 2ZI2; -.
DR PDBsum; 2ZIQ; -.
DR PDBsum; 2ZNK; -.
DR PDBsum; 2ZO3; -.
DR PDBsum; 3D49; -.
DR PDBsum; 3DHK; -.
DR PDBsum; 3DT0; -.
DR PDBsum; 3DUX; -.
DR PDBsum; 3EGK; -.
DR PDBsum; 3EQ0; -.
DR PDBsum; 3LDX; -.
DR PDBsum; 3UTU; -.
DR PDBsum; 4AX9; -.
DR PDBsum; 4BAH; -.
DR PDBsum; 4BAK; -.
DR PDBsum; 4BAM; -.
DR PDBsum; 4BAN; -.
DR PDBsum; 4BAQ; -.
DR PDBsum; 4HIR; -.
DR PDBsum; 4LXB; -.
DR PDBsum; 4MLF; -.
DR PDBsum; 4YES; -.
DR PDBsum; 5HIR; -.
DR PDBsum; 6HIR; -.
DR PDBsum; 7A0D; -.
DR PDBsum; 7A0E; -.
DR PDBsum; 7A0F; -.
DR AlphaFoldDB; P01050; -.
DR BMRB; P01050; -.
DR SMR; P01050; -.
DR Allergome; 9843; Hir me Hirudin.
DR MEROPS; I14.001; -.
DR EvolutionaryTrace; P01050; -.
DR GO; GO:0005615; C:extracellular space; IDA:CAFA.
DR GO; GO:0004867; F:serine-type endopeptidase inhibitor activity; IDA:CAFA.
DR GO; GO:1902572; P:negative regulation of serine-type peptidase activity; IDA:CAFA.
DR DisProt; DP00137; -.
DR Gene3D; 2.70.10.10; -; 1.
DR InterPro; IPR024793; Hirudin.
DR InterPro; IPR011061; Hirudin/antistatin.
DR InterPro; IPR000429; Prot_inh_hirudin.
DR Pfam; PF00713; Hirudin; 1.
DR PIRSF; PIRSF001640; Hirudin; 1.
DR PRINTS; PR00777; HIRUDIN.
DR SUPFAM; SSF57262; SSF57262; 1.
PE 1: Evidence at protein level;
KW 3D-structure; Direct protein sequencing; Disulfide bond; Glycoprotein;
KW Pharmaceutical; Protease inhibitor; Secreted; Serine protease inhibitor;
KW Sulfation.
FT CHAIN 1..65
FT /note="Hirudin variant-1"
FT /id="PRO_0000195639"
FT REGION 1..3
FT /note="Interaction with thrombin active site"
FT REGION 39..65
FT /note="Disordered"
FT /evidence="ECO:0000256|SAM:MobiDB-lite"
FT REGION 55..65
FT /note="Interaction with fibrinogen-binding exosite of
FT thrombin"
FT MOD_RES 63
FT /note="Sulfotyrosine"
FT /evidence="ECO:0000269|PubMed:17685615,
FT ECO:0000269|PubMed:8251938"
FT CARBOHYD 45
FT /note="O-linked (GalNAc...) threonine"
FT /evidence="ECO:0000250"
FT DISULFID 6..14
FT DISULFID 16..28
FT DISULFID 22..39
FT MUTAGEN 1
FT /note="V->E,K: Strongly decreased affinity for thrombin."
FT /evidence="ECO:0000269|PubMed:1581311"
FT MUTAGEN 1
FT /note="V->G,S: Decreased affinity for thrombin."
FT /evidence="ECO:0000269|PubMed:1581311"
FT MUTAGEN 1
FT /note="V->L,R: No effect."
FT /evidence="ECO:0000269|PubMed:1581311"
FT MUTAGEN 2
FT /note="V->E,G: Strongly decreased affinity for thrombin."
FT /evidence="ECO:0000269|PubMed:1581311"
FT MUTAGEN 2
FT /note="V->L: Decreased affinity for thrombin."
FT /evidence="ECO:0000269|PubMed:1581311"
FT MUTAGEN 3
FT /note="Y->A: Strongly decreased affinity for thrombin."
FT /evidence="ECO:0000269|PubMed:1581311"
FT MUTAGEN 4
FT /note="T->A: Decreased affinity for thrombin."
FT /evidence="ECO:0000269|PubMed:1581311"
FT MUTAGEN 5
FT /note="D->A,E: Decreased affinity for thrombin."
FT /evidence="ECO:0000269|PubMed:1581311"
FT MUTAGEN 15
FT /note="L->A: Strongly decreased affinity for thrombin."
FT /evidence="ECO:0000269|PubMed:8135762"
FT MUTAGEN 17
FT /note="E->A: Slightly decreased affinity for thrombin."
FT MUTAGEN 20
FT /note="N->A: Decreased affinity for thrombin."
FT /evidence="ECO:0000269|PubMed:8135762"
FT MUTAGEN 21
FT /note="V->A: Slightly decreased affinity for thrombin."
FT /evidence="ECO:0000269|PubMed:8135762"
FT MUTAGEN 56
FT /note="F->A,W: Slightly decreased affinity for thrombin."
FT /evidence="ECO:0000269|PubMed:1911777"
FT MUTAGEN 56
FT /note="F->I,L,T,V: Strongly decreased affinity for
FT thrombin."
FT /evidence="ECO:0000269|PubMed:1911777"
FT MUTAGEN 56
FT /note="F->Y: No effect."
FT /evidence="ECO:0000269|PubMed:1911777"
FT MUTAGEN 60
FT /note="P->A,G: Decreased affinity for thrombin."
FT /evidence="ECO:0000269|PubMed:1911777"
FT MUTAGEN 63
FT /note="Y->A,E,L: Slightly decreased affinity for thrombin."
FT /evidence="ECO:0000269|PubMed:1911777"
FT MUTAGEN 63
FT /note="Y->F: No effect."
FT /evidence="ECO:0000269|PubMed:1911777"
FT MUTAGEN 63
FT /note="Y->V: Decreased affinity for thrombin."
FT /evidence="ECO:0000269|PubMed:1911777"
FT STRAND 11..15
FT /evidence="ECO:0007829|PDB:2PW8"
FT STRAND 17..19
FT /evidence="ECO:0007829|PDB:2PW8"
FT STRAND 20..22
FT /evidence="ECO:0007829|PDB:4HIR"
FT STRAND 26..29
FT /evidence="ECO:0007829|PDB:2PW8"
FT STRAND 32..34
FT /evidence="ECO:0007829|PDB:1HIC"
FT STRAND 38..42
FT /evidence="ECO:0007829|PDB:2PW8"
FT HELIX 61..63
FT /evidence="ECO:0007829|PDB:2UUF"
SQ SEQUENCE 65 AA; 6970 MW; 9085A5876E3DE9FF CRC64;
VVYTDCTESG QNLCLCEGSN VCGQGNKCIL GSDGEKNQCV TGEGTPKPQS HNDGDFEEIP
EEYLQ
