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Information on EC 3.4.24.72 - fibrolase and Organism(s) Agkistrodon contortrix contortrix and UniProt Accession P28891

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EC Tree
     3 Hydrolases
         3.4 Acting on peptide bonds (peptidases)
             3.4.24 Metalloendopeptidases
                3.4.24.72 fibrolase
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Agkistrodon contortrix contortrix
UNIPROT: P28891 not found.
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Word Map
The taxonomic range for the selected organisms is: Agkistrodon contortrix contortrix
The expected taxonomic range for this enzyme is: Eukaryota, Bacteria
Synonyms
fibrolase, alfimeprase, fibrinolytic protease, neuwiedase, brevithrombolase, starase, fibrinolytic proteinase, ufeiii, snake venom fibrinolytic enzyme, cfr15-protease, more
SYNONYM
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
snake venom fibrinolytic enzyme
-
3-203-fibrolase[3-serine]
-
-
Agkistrodon contortrix contortrix metalloproteinase
-
-
-
-
Alfimeprase
Fibrinolytic proteinase
-
-
-
-
Non-hemorrhagic fibrinolytic metalloproteinase
-
-
-
-
Proteinase, Agkistrodon contortrix contortrix venom metallo-
-
-
-
-
VlF
-
-
-
-
REACTION TYPE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
hydrolysis of peptide bond
-
-
-
-
CAS REGISTRY NUMBER
COMMENTARY hide
116036-70-5
-
SUBSTRATE
PRODUCT                       
REACTION DIAGRAM
ORGANISM
UNIPROT
COMMENTARY
(Substrate) hide
LITERATURE
(Substrate)
COMMENTARY
(Product) hide
LITERATURE
(Product)
Reversibility
r=reversible
ir=irreversible
?=not specified
Fibrin + H2O
?
show the reaction diagram
Fibrinogen + H2O
?
show the reaction diagram
the enzyme has a direct proteolytic activity against the A alpha-chain of fibrinogen at positions Lys413 and Leu414
-
-
?
fibrinogen + H2O
fibrin + propeptide
show the reaction diagram
4-amino-benzene-His-Thr-Glu-Lys-leu-Val-Thr-Ser-dinitrophenol + H2O
?
show the reaction diagram
-
-
-
-
?
azocasein + H2O
?
show the reaction diagram
-
-
-
-
?
bradykinin + H2O
Arg-Pro-Pro-Gly-Phe-Ser-Pro + Phe-Arg
show the reaction diagram
-
-
-
-
?
Fibrin + H2O
?
show the reaction diagram
Fibrin Aalpha-chain + H2O
?
show the reaction diagram
-
human, cleavage at Lys413-Lys414, the Bbeta-chain is cleaved more slowly and the gamma-chain is minimally affected
-
-
?
Fibrinogen + H2O
?
show the reaction diagram
-
fibrolase rapidly cleaves the A(alpha)-chain of fibrinogen and the B(beta)-chain at a slower rate, while it has no activity on the gamma-chain. The primary cleavage site at the alpha-chain ist he Lys-Leu bond at residues 413-414
-
-
?
fibrinogen + H2O
fibrin + propeptide
show the reaction diagram
Fibrinogen Aalpha-chain + H2O
?
show the reaction diagram
His-Thr-Glu-Ala-Leu-Val-Thr-Ser + H2O
?
show the reaction diagram
-
-
-
-
?
His-Thr-Glu-Arg-Leu-Val-Thr-Ser + H2O
?
show the reaction diagram
-
-
-
-
?
His-Thr-Glu-Asn-Leu-Val-Thr-Ser + H2O
?
show the reaction diagram
-
-
-
-
?
His-Thr-Glu-Glu-Leu-Val-Thr-Ser + H2O
?
show the reaction diagram
-
-
-
-
?
His-Thr-Glu-His-Leu-Val-Thr-Ser + H2O
?
show the reaction diagram
-
-
-
-
?
His-Thr-Glu-Leu-Leu-Val-Thr-Ser + H2O
?
show the reaction diagram
-
-
-
-
?
His-Thr-Glu-Lys-Leu-Val-Thr-Ser + H2O
?
show the reaction diagram
-
cleaving sequence residues 410-417 from fibrinogen Aalpha-chain
-
-
?
His-Thr-Glu-Phe-Leu-Val-Thr-Ser + H2O
?
show the reaction diagram
-
-
-
-
?
His-Thr-Glu-Pro-Leu-Val-Thr-Ser + H2O
?
show the reaction diagram
-
-
-
-
?
His-Thr-Glu-Ser-Leu-Val-Thr-Ser + H2O
?
show the reaction diagram
-
-
-
-
?
Insulin B-chain + H2O
?
show the reaction diagram
Leu-Val-Glu-Ala-Leu-Tyr-Leu-Val + H2O
?
show the reaction diagram
-
cleaving sequence residues 11-18 from insulin B-chain
-
-
?
low molecular weight kininogen + H2O
kallidin + ?
show the reaction diagram
-
-
i.e. Lys-Arg-Pro-Pro-Gly-Phe-Ser-Pro-Phe-Arg
-
?
low-molecular weight kininogen + H2O
kallidin + ?
show the reaction diagram
additional information
?
-
NATURAL SUBSTRATE
NATURAL PRODUCT
REACTION DIAGRAM
ORGANISM
UNIPROT
COMMENTARY
(Substrate) hide
LITERATURE
(Substrate)
COMMENTARY
(Product) hide
LITERATURE
(Product)
REVERSIBILITY
r=reversible
ir=irreversible
?=not specified
Fibrin + H2O
?
show the reaction diagram
Fibrinogen + H2O
?
show the reaction diagram
the enzyme has a direct proteolytic activity against the A alpha-chain of fibrinogen at positions Lys413 and Leu414
-
-
?
fibrinogen + H2O
fibrin + propeptide
show the reaction diagram
bovine substrate
-
-
?
bradykinin + H2O
Arg-Pro-Pro-Gly-Phe-Ser-Pro + Phe-Arg
show the reaction diagram
-
-
-
-
?
Fibrin + H2O
?
show the reaction diagram
Fibrinogen + H2O
?
show the reaction diagram
-
fibrolase rapidly cleaves the A(alpha)-chain of fibrinogen and the B(beta)-chain at a slower rate, while it has no activity on the gamma-chain. The primary cleavage site at the alpha-chain ist he Lys-Leu bond at residues 413-414
-
-
?
fibrinogen + H2O
fibrin + propeptide
show the reaction diagram
-
fibrinogenolytic activity
-
-
?
low molecular weight kininogen + H2O
kallidin + ?
show the reaction diagram
-
-
i.e. Lys-Arg-Pro-Pro-Gly-Phe-Ser-Pro-Phe-Arg
-
?
low-molecular weight kininogen + H2O
kallidin + ?
show the reaction diagram
-
initiates generation of bradykinin
-
-
?
additional information
?
-
METALS and IONS
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
Calcium
-
potential binding site for calcium
INHIBITOR
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
1,10-phenanthroline
alpha2-Macroglobulin
-
tetraethylene pentamine
-
rapid and complete inhibition
Tetraethylenepentamine
-
-
additional information
-
ACTIVATING COMPOUND
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
additional information
-
Ki VALUE [mM]
INHIBITOR
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
additional information
additional information
-
native and PEG-bound enzyme, rapid interaction kinetics with inhibitor alpha2-macroglobulin
-
SPECIFIC ACTIVITY [µmol/min/mg]
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
10900
-
purified PEG-bound enzyme, substrate human fibrin
11300
-
purified native enzyme, substrate human fibrin
1200
-
purified PEG-bound enzyme, substrate azocasein
1260
-
purified native enzyme, substrate azocasein
additional information
pH OPTIMUM
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
TEMPERATURE OPTIMUM
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
pI VALUE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
ORGANISM
COMMENTARY hide
LITERATURE
UNIPROT
SEQUENCE DB
SOURCE
UNIPROT
ENTRY NAME
ORGANISM
NO. OF AA
NO. OF TRANSM. HELICES
MOLECULAR WEIGHT[Da]
SOURCE
SEQUENCE
LOCALIZATION PREDICTION?
VM1F_AGKCO
203
0
22908
Swiss-Prot
Mitochondrion (Reliability: 5)
MOLECULAR WEIGHT
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
23000
23800
x * 23800, recombinant His-tagged alfimeprase, SDS-PAGE
22540
-
Agkistrodon contortrix, calculation from amino acid sequence
22750
-
Agkistrodon contortrix, isoform fib2, calculation from amino acid sequence
22880
-
Agkistrodon contortrix, isoform fib1, calculation from amino acid sequence
23000
43330
-
PEG-bound enzyme, gel filtration
60000
-
x * 23000, native enzyme, x * 60000, PEG-bound enzyme
additional information
-
amino acid sequence
SUBUNIT
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
additional information
POSTTRANSLATIONAL MODIFICATION
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
additional information
-
contains little or no carbohydrate
CRYSTALLIZATION (Commentary)
ORGANISM
UNIPROT
LITERATURE
PROTEIN VARIANTS
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
additional information
-
construction of a heterobifunctional chimeric derivative of fibrolase through linking covalently with a RGD-like peptide via a surface residue distant from the active site, structure, overview, the chimera shows fibrolase and platelet-binding ability, the chimeric enzyme exhibits an IC50 of 105 nM in inhibition of platelet aggregation, overview
pH STABILITY
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
5.5
-
6 h, 30-35% loss of activity
31471
6.5
-
optimal pH for stability
31471
6.5 - 10
-
stable
31481
9
-
6 h, 45-50% loss of activity
31471
additional information
-
effect of pH on conformation
31481
TEMPERATURE STABILITY
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
22
-
stable for 10 days
37
-
about 50% loss of activity after 2 days, complete loss of activity after 10 days
additional information
-
effect of temperature on conformation
GENERAL STABILITY
ORGANISM
UNIPROT
LITERATURE
Effect of zinc binding on the structure and stability of fibrolase
-
STORAGE STABILITY
ORGANISM
UNIPROT
LITERATURE
22°C, stable for 10 days
-
4°C or -20°C, stable for 6 months, 20-30% loss of activity after 1 year
-
PURIFICATION (Commentary)
ORGANISM
UNIPROT
LITERATURE
HiTrap Q-Sepharose column chromatography, gel filtration
recombinant soluble His-tagged enzyme from Escherichia coli by nickel affinity and hydrophobic interaction chromatography to 95.6% purity
secreted recombinant His-tagged alfimeprase from Pichia pastoris strains of GS115 and KM71 cell culture medium by immunoaffinity chromatography to 96% purity
hydrophobic interaction HPLC, hydroxyapatite HPLC and cation exchange HPLC
-
immunoaffinity chromatography
-
native enzyme by hydrophobic interaction and hydroxyapatite chromatography
-
CLONED (Commentary)
ORGANISM
UNIPROT
LITERATURE
chimeric Arg-Gly-Asp-fibrolase is expressed in Escherichia coli BL21(DE3) cells
co-expression of the active signal-less His-tagged enzyme with chaperone DsbC in Escherichia coli cytoplasm, over 90% of the recombinant enzyme is soluble due to DsbC, while the chaperones FkpA and Skp do not have an effect on fibrolase solubility
gene alf, high level expression of His-tagged alfimeprase in Pichia pastoris strains of GS115 and KM71 by methanol induction, secretion of the recombinant enzyme to the cell culture medium
expressed in Pichia pastoris
-
expression of the wild-type fibrolase and a truncated enzyme form, alfimeprase
-
production of alfimeprase by recombinant DNA technology
-
RENATURED/Commentary
ORGANISM
UNIPROT
LITERATURE
using 50 mM Tris-HCl, 0.5 M L-arginine, 1 M NaCl, 0.5 mM ZnCl2, 1 mM oxidized glutathione, 2 mM reduced glutathione, and 2 M urea at pH 7.0. After the protein is added, the pH is adjusted to 7.5, and the protein is left to refold at 4°C for approximately 24 h
APPLICATION
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
medicine
development of more effective therapeutic approaches for treating occlusive thrombotic diseases, potential clinical interest in fibrolase for pharmacologic dissolution of an established thrombus
drug development
-
summary of results (including adverse events and pharmacodynamics) regarding drug development of alfimeprase as a potential drug against thrombosis and vascular disorders
medicine
-
the use of the recombinant fibrolase alfimeprase results in rapid restoration of arterial patency in less than 4 h in most peripheral arterial occlusion patients
pharmacology
-
alfimeprase is a potential therapeutic agents in thrombosis therapy, clinical indications and studies, overview
REF.
AUTHORS
TITLE
JOURNAL
VOL.
PAGES
YEAR
ORGANISM (UNIPROT)
PUBMED ID
SOURCE
Guan, A.L.; Retzios, A.D.; Henderson, G.N.; Markland, F.S.
Purification and characterization of a fibrinolytic enzyme from venom of the southern copperhead snake (Agkistrodon contortrix contortrix)
Arch. Biochem. Biophys.
289
197-207
1991
Agkistrodon contortrix contortrix
Manually annotated by BRENDA team
Retzios, A.D.; Markland, F.S.
A direct-acting fibrinolytic enzyme from the venom of Agkistrodon contortrix contortrix: effects on various components of the human blood coagulation and fibrinolysis systems
Thromb. Res.
52
541-552
1988
Agkistrodon contortrix contortrix
Manually annotated by BRENDA team
Randolph, A.; Chamberlain, S.H.; Chu, H.L.C.; Retzios, A.D.; Markland, F.S.; Masiarz, F.R.
Amino acid sequence of fibrolase, a direct-acting fibrinolytic enzyme from Agkistrodon contortrix contortrix venom
Protein Sci.
1
590-600
1992
Agkistrodon contortrix contortrix
Manually annotated by BRENDA team
Loayza, S.L.; Trikha, M.; Markland, F.S.; Riquelme, P.; Kuo, J.
Resolution of isoforms of natural and recombinant fibrolase, the fibrinolytic enzyme from Agkistrodon contortrix contortrix snake venom, and comparison of their EDTA sensitivities
J. Chromatogr.
662
227-243
1994
Agkistrodon contortrix contortrix
Manually annotated by BRENDA team
Retzios, A.D.; Markland, F.S.
Fibrinolytic enzymes from the venoms of Agkistrodon contortrix contortrix and Crotalus basiliscus basiliscus: cleavage site specificity towards the alpha-chain of fibrin
Thromb. Res.
74
355-367
1994
Agkistrodon contortrix contortrix
Manually annotated by BRENDA team
Manning, M.C.
Sequence analysis of fibrolase, a fibrinolytic metalloproteinase from Agkistrodon contortrix contortrix
Toxicon
33
1189-1200
1995
Agkistrodon contortrix contortrix
Manually annotated by BRENDA team
Trikha, M.; Schmitmeier, S.; Markland, F.S.
Purification and characterization of fibrolase isoforms from venom of individual southern copperhead (Agkistrodon contortrix Contortrix) snakes
Toxicon
32
1521-1531
1994
Agkistrodon contortrix contortrix
Manually annotated by BRENDA team
Patton, L.M.; Pretzer, D.; Schulteis, B.S.; Saggart, B.S.; Tennant, K.D.; Ahmed, N.K.
Activity assays for characterizing the thrombolytic protein fibrolase
J. Biochem. Biophys. Methods
27
11-23
1993
Agkistrodon contortrix contortrix
Manually annotated by BRENDA team
Markland, F.S.; Morris, S.; Deschamps, J.R.; Ward, K.B.
Resolution of isoforms of natural and recombinant fibrinolytic snake venom enzyme using high performance capillary electrophoresis
J. Liq. Chromatogr.
16
2189-2201
1993
Agkistrodon contortrix contortrix
-
Manually annotated by BRENDA team
Pretzer, D.; Schulteis, B.; Vander Velde, D.G.; Smith, C.D.; Mitchell, J.W.; Manning, M.C.
Effect of zinc binding on the structure and stability of fibrolase, a fibrinolytic protein from snake venom
Pharm. Res.
9
870-877
1992
Agkistrodon contortrix contortrix
Manually annotated by BRENDA team
Pretzer, D.; Schulteis, B.S.; Smith, C.D.; Vander Velde, D.G.; Mitchell, J.W.; Manning, M.C.
Stability of the thrombolytic protein fibrolase: effect of temperature and pH on activity and conformation
Pharm. Res.
8
1103-1112
1991
Agkistrodon contortrix contortrix
Manually annotated by BRENDA team
Chen, H.M.; Guan, A.L.; Markland, F.S.
Immunological properties of the fibrinolytic enzyme (fibrolase) from southern copperhead (Agkistrodon contortrix contortrix) venom and its purification by immunoaffinity chromatography
Toxicon
29
683-694
1991
Agkistrodon contortrix contortrix
Manually annotated by BRENDA team
Guan, A.L.; Markland, F.S.
Isoelectric focusing in immobilized pH gradients of a snake venom fibrinolytic enzyme
J. Biochem. Biophys. Methods
16
215-226
1988
Agkistrodon contortrix contortrix
Manually annotated by BRENDA team
Bolger, M.B.; Swenson, S.; Markland, F.S., Jr.
Three-dimensional structure of fibrolase, the fibrinolytic enzyme from southern copperhead venom, modeled from the X-ray structure of adamalysin II and atrolysin C
AAPS Pharmsci.
3
E16
2001
Agkistrodon contortrix contortrix (P28891), Agkistrodon contortrix contortrix
Manually annotated by BRENDA team
Shi, J.; Zhang, S.T.; Zhang, X.J.; Xu, H.; Guo, A.G.
Expression, purification, and activity identification of alfimeprase in Pichia pastoris
Protein Expr. Purif.
54
240-246
2007
Agkistrodon contortrix contortrix (P28891)
Manually annotated by BRENDA team
Zhang, S.T.; Shi, J.; Zhao, J.; Qi, Y.F.; Guo, A.G.
Expression of soluble and functional snake venom fibrinolytic enzyme fibrolase via the co-expression of DsbC in Escherichia coli
Protein Pept. Lett.
13
559-563
2006
Agkistrodon contortrix contortrix (P28891), Agkistrodon contortrix contortrix
Manually annotated by BRENDA team
Swenson, S.; Markland, F.S.
Fibrolase
Toxin Rev.
25
351-378
2006
Agkistrodon contortrix contortrix
-
Manually annotated by BRENDA team
Toombs, C.F.; Deitcher, S.R.
Nonclinical and clinical characterization of a novel acting thrombolytic: alfimeprase
Toxin Rev.
25
379-392
2006
Agkistrodon contortrix contortrix
-
Manually annotated by BRENDA team
No authors listed
Alfimeprase
Drugs R. D.
9
185-190
2008
Agkistrodon contortrix contortrix
Manually annotated by BRENDA team
Fang, H.M.; Zhao, L.; Lu, P.; Chen, S.J.; Bao, Z.X.; Qin, Y.F.; Zhu, Z.Y.; Zhao, J.M.; Mai, J.; Zhang, S.T.
Modular design, expression and characterization of novel bifunctional mutants of fibrolase with combined platelet aggregation-inhibition and fibrinolytic activity
Protein J.
30
247-252
2011
Agkistrodon contortrix contortrix (P28891), Agkistrodon contortrix contortrix
Manually annotated by BRENDA team
Markland, F.S.; Swenson, S.
Fibrolase: trials and tribulations
Toxins
2
793-808
2010
Agkistrodon contortrix contortrix
Manually annotated by BRENDA team