Information on EC 3.4.24.B33 - Vipera ammodytes ammodytes metalloproteinase VaH3

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The expected taxonomic range for this enzyme is: Vipera ammodytes ammodytes

EC NUMBER
COMMENTARY hide
3.4.24.B33
preliminary BRENDA-supplied EC number
RECOMMENDED NAME
GeneOntology No.
Vipera ammodytes ammodytes metalloproteinase VaH3
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REACTION
REACTION DIAGRAM
COMMENTARY hide
ORGANISM
UNIPROT
LITERATURE
Cleavage of the Lys413-/-Leu414 bond of alpha-chain of human fibrinogen. Cleavage of Ala14-/-Leu15 and more slowly Tyr16-/-Leu17 in insulin B chain. Hemorrhagic metalloproteinase
show the reaction diagram
ORGANISM
COMMENTARY hide
LITERATURE
UNIPROT
SEQUENCE DB
SOURCE
GENERAL INFORMATION
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
evolution
the enzyme VaH3 belongs to the P-IIIc class of snake venom metalloproteinases
physiological function
Zn2+-dependent metalloproteinases play a major part in the pathological effects of viperid snake bites, the most pronounced being local and systemic hemorrhage, local tissue damage and coagulopathy. Enzyme VaH3 is one of the principal hemorrhagins in Vipera ammodytes ammodytes venom The enzyme is an effective alpha-fibrinogenase that cleaves prothrombin and factor X without activating them. VaH3 only very weakly inhibits collagen-, ADP- and ristocetin-induced platelet aggregation
additional information
enzyme three-dimensional structure model and structure-function relationship analysis, overview
SUBSTRATE
PRODUCT                       
REACTION DIAGRAM
ORGANISM
UNIPROT
COMMENTARY
(Substrate) hide
LITERATURE
(Substrate)
COMMENTARY
(Product) hide
LITERATURE
(Product)
Reversibility
r=reversible
ir=irreversible
?=not specified
bovine factor X + H2O
?
show the reaction diagram
the enzyme is able to activate factor X only to a very small extent. However it strongly degrades factor X. The major proteolytic products accumulates between 34 and 37 kDa and their N-terminals correspond to cleavage at residues 17, 20 and 22 upstream of the N-terminal of the factor Xa heavy chain
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?
bovine fibronectin + H2O
?
show the reaction diagram
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?
bovine prothrombin + H2O
?
show the reaction diagram
the enzyme degrades prothrombin in vitro, however in a nonactivating way. VaH3 cleaves the molecule at sites involved in the physiological process of its activation. This results in the formation of prethrombin-1 and prethrombin-2, along with fragments 1 and 2. However, VaH3 does not cleave the Arg320-/-Ile321 peptide bond that is essential for the formation of active alpha-thrombin
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?
Collagen IV + H2O
?
show the reaction diagram
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?
factor X + H2O
?
show the reaction diagram
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?
fibrinogen alpha-chain + H2O
?
show the reaction diagram
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?
Fibronectin + H2O
?
show the reaction diagram
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?
human fibrinogen alpha-chain + H2O
?
show the reaction diagram
the alpha-chain of human fibrinogen is cleaved between Lys413 and Leu414, no hydrolysis of the beta- or gamma-chains is observed
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?
human type collagen IV + H2O
?
show the reaction diagram
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?
Insulin B-chain + H2O
?
show the reaction diagram
VaH3 rapidly cleaves the peptide bond Ala14-/-Leu15, the bond Tyr16-/-Leu17 is hydrolyzed at a much slower rate
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?
murine laminin + H2O
?
show the reaction diagram
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?
murine nidogen + H2O
?
show the reaction diagram
from Matrigel Growth Factor Reduced, preferentially cleaved at positions Ser322-/-Phe323 and and Tyr352-/-Asn353
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?
Nidogen + H2O
?
show the reaction diagram
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?
additional information
?
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NATURAL SUBSTRATES
NATURAL PRODUCTS
REACTION DIAGRAM
ORGANISM
UNIPROT
COMMENTARY
(Substrate) hide
LITERATURE
(Substrate)
COMMENTARY
(Product) hide
LITERATURE
(Product)
REVERSIBILITY
r=reversible
ir=irreversible
?=not specified
Collagen IV + H2O
?
show the reaction diagram
R4NNL0
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?
factor X + H2O
?
show the reaction diagram
R4NNL0
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?
fibrinogen alpha-chain + H2O
?
show the reaction diagram
R4NNL0
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?
Fibronectin + H2O
?
show the reaction diagram
R4NNL0
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?
Nidogen + H2O
?
show the reaction diagram
R4NNL0
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?
additional information
?
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R4NNL0
hydrolyzes plasma proteins involved in blood coagulation. VaH3 only very weakly inhibits collagen-, ADP- and ristocetin-induced platelet aggregation
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METALS and IONS
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
Ca2+
required for enzyme stability and activity
Zn2+
dependent on, required for enzyme stability
INHIBITORS
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
anti-ammodytagin antibodies
complete inhibition, the antibodies strongly crossreact with VaH3 and completely neutralize its hemorrhagic activity in rat
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EDTA
alpha-fibrinogenolytic activity is completely inhibited
additional information
no inhibition by PMSF
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pH OPTIMUM
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
TEMPERATURE OPTIMUM
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
pI VALUE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
6.2
isoelectric focusing
SOURCE TISSUE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
SOURCE
LOCALIZATION
ORGANISM
UNIPROT
COMMENTARY hide
GeneOntology No.
LITERATURE
SOURCE
MOLECULAR WEIGHT
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
53700
2 * 58000, SDS-PAGE, 2 * 53700, mass spectrometric analysis after chemical reduction and S-carbamoylmethylation
58000
2 * 58000, SDS-PAGE, 2 * 53700, mass spectrometric analysis after chemical reduction and S-carbamoylmethylation
104000
MALDI/TOF mass spectrometric analysis
SUBUNITS
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
homodimer
2 * 58000, SDS-PAGE, 2 * 53700, mass spectrometric analysis after chemical reduction and S-carbamoylmethylation
additional information
two identical, covalently linked subunits, each of the identical glycoprotein subunits comprise a metalloproteinase, a disintegrin-like domain and a cysteine-rich domain. Enzyme three-dimensional structure modeling and structure-function relationship analysis, peptide mapping after digestion of the enzyme by endoproteinase Lys-C, overview
POSTTRANSLATIONAL MODIFICATION
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
glycoprotein
low N-glycosylation level, deglycosylation reduces the molecular weight by 4.6 kDa
GENERAL STABILITY
ORGANISM
UNIPROT
LITERATURE
enzyme VaH3 is a labile protein that rapidly loses its proteolytic and hemorrhagic activities, presence of imidazole and absence of Ca2+ ions in the buffers reduces VaH3 stability, also 50 mM NaCl or KCl reduce the enzyme stability
stability depends on the presence of Zn2+ and Ca2+ ions
Purification/COMMENTARY
ORGANISM
UNIPROT
LITERATURE
native enzyme from venom by gel filtration, concanavalin A affinity and anion-exchange chromatography, followed by hydroxyapatite and cation exchange chromatography, homogenization by isoelectric focusing
Cloned/COMMENTARY
ORGANISM
UNIPROT
LITERATURE
DNA and amino acid sequence determination and analysis
APPLICATION
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
medicine
snake venom metalloproteinases are important targets in antivenom therapy