Information on EC 3.4.24.87 - ADAMTS13 endopeptidase

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The enzyme appears in viruses and cellular organisms

EC NUMBER
COMMENTARY hide
3.4.24.87
-
RECOMMENDED NAME
GeneOntology No.
ADAMTS13 endopeptidase
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REACTION
REACTION DIAGRAM
COMMENTARY hide
ORGANISM
UNIPROT
LITERATURE
The enzyme cleaves the von Willebrand factor at bond Tyr842-/-Met843 within the A2 domain
show the reaction diagram
REACTION TYPE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
CAS REGISTRY NUMBER
COMMENTARY hide
334869-10-2
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ORGANISM
COMMENTARY hide
LITERATURE
UNIPROT
SEQUENCE DB
SOURCE
beagle dogs
-
-
Manually annotated by BRENDA team
Dunkin Hartley guinea pigs
-
-
Manually annotated by BRENDA team
cynomolgus monkey
-
-
Manually annotated by BRENDA team
BALB/c
-
-
Manually annotated by BRENDA team
New Zealand white rabbits
-
-
Manually annotated by BRENDA team
Yorkshire strain
-
-
Manually annotated by BRENDA team
GENERAL INFORMATION
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
malfunction
metabolism
-
cyclophilin B knock-out mice have reduced ADAMTS13 levels
physiological function
additional information
SUBSTRATE
PRODUCT                       
REACTION DIAGRAM
ORGANISM
UNIPROT
COMMENTARY
(Substrate) hide
LITERATURE
(Substrate)
COMMENTARY
(Product) hide
LITERATURE
(Product)
Reversibility
r=reversible
ir=irreversible
?=not specified
Collagen + H2O
?
show the reaction diagram
-
-
-
-
?
DREQAPNLVYMVTGNPASDEIKRLPGDIQVVPIGVGPNANVQELERIGWPNAPILIQDFETLPREAPDLVLQRA + H2O
?
show the reaction diagram
-
i.e. VWF74 peptide, a pseudo-wild-type peptide von Willebrand factor 74, VWF74, encompassing the von Willebrand factor, VWF, A2 domain sequence 1596-1669
-
-
?
fluorescence resonance energy transfer substrate-von Willebrand factor 73 + H2O
?
show the reaction diagram
-
-
-
-
?
fluorescent resonance energy transfer-von Willebrand factor 73 + H2O
?
show the reaction diagram
-
-
-
-
?
FRET-VWF115 peptide + H2O
?
show the reaction diagram
-
von Willebrand factor-derived peptide substrate comprising residues 1554-1668
-
-
?
FRET-VWF73 + H2O
?
show the reaction diagram
-
fluorogenic von Willebrand factor-derived peptide substrate. The distal C-terminal domains of ADAMTS13 are not necessary for the cleavage of the VWF73-based peptide substrate
-
-
?
FRETS-rVWF71 + H2O
?
show the reaction diagram
-
substrate based on von Willebrand factor residues Gln1599-Arg1668, with an N-terminal Gly and with mutations N1610C and K1617R. The N-terminus is modified with IRDye QC-1 Nhydroxysuccinimide ester, and Cys1610 is modified with DyLight 633 maleimide
-
-
?
FRETS-von Willebrand factor 73 + H2O
?
show the reaction diagram
-
-
-
-
?
FRETS-vWF73 + H2O
?
show the reaction diagram
-
a fluorogenic von Willebrand factor-derived peptide substrate
-
-
?
FRETS-VWF73 peptide + H2O
?
show the reaction diagram
-
fluorogenic von Willebrand factor-derived peptide substrate
-
-
?
FRETSVWF73 + H2O
?
show the reaction diagram
-
a von Willebrand factor-derived fluorescein-labeled peptide substrate
-
-
?
FRETSVWF73 peptide + H2O
?
show the reaction diagram
-
a von Willebrand factor-derived fluorescein-labeled peptide substrate
-
-
?
GST-von Willebrand factor 73 + H2O
?
show the reaction diagram
-
contains residues Asp1596-Arg1668 from von Willebrand factor domain A2
-
-
?
GST-VWF73 + H2O
?
show the reaction diagram
HRPH-A2-B
?
show the reaction diagram
-
HRPH-A2-B is a derivative of von Willebrand factor 73, consisting of a HRP conjugate of a biotinylated von Willebrand factor 78 sequence
cleavage of Tyr842-Met843 within the A2 domain, i.e. Tyr1605-Met1606 in von Willebrand factor UniProt Id P04275
-
-
large von Willebrand factor multimer + H2O
?
show the reaction diagram
-
-
-
-
?
proteins + H2O
peptides
show the reaction diagram
ultra-large von Willebrand factor + H2O
?
show the reaction diagram
-
-
-
-
?
ultra-large von Willebrand factor multimer + H2O
?
show the reaction diagram
-
-
-
-
?
von Willebrand factor + H2O
2 peptides
show the reaction diagram
von Willebrand factor + H2O
2 peptides of 140 kD and 65 kD
show the reaction diagram
-
cleavage of peptide bond Tyr842-Met843
-
?
von Willebrand factor + H2O
2 peptides of 140 kDa and 176 kDa
show the reaction diagram
von Willebrand factor + H2O
2 peptides of 176 kD and 140 kD
show the reaction diagram
von Willebrand factor + H2O
?
show the reaction diagram
von Willebrand factor + H2O
von Willebrand factor 140-kD fragment + von Willebrand factor 176-kD fragment
show the reaction diagram
von Willebrand factor + H2O
von Willebrand factor fragments
show the reaction diagram
von Willebrand factor 115 (1554-1668) + H2O
?
show the reaction diagram
-
A2 domain fragment
-
-
?
von Willebrand factor 115 + H2O
10000 Da fragment of von Willebrand factor 115 + 7000 Da fragment of von Willebrand factor 115
show the reaction diagram
-
von Willebrand factor is cleaved at the Tyr1605-Met1606 bond in the von Willebrand factor A2 domain
-
-
?
von Willebrand factor 115-A3 (1554-1874) + H2O
?
show the reaction diagram
-
A2 domain fragment
-
-
?
von Willebrand factor 73 + H2O
7722 Da peptide + ?
show the reaction diagram
-
-
-
-
?
von Willebrand factor 73 + H2O
?
show the reaction diagram
-
minimal substrate cleavable by ADAMTS-13
-
-
?
von Willebrand factor 76 (1593-1668) + H2O
?
show the reaction diagram
-
A2 domain fragment
-
-
?
VWF115 + H2O
10 kDa VWF115 fragment + 7 kDa VWF115 fragment
show the reaction diagram
-
VWFA2 domain fragment, spanning von Willebrand factor residues 1554-1668, generation of 2 cleavage products of 10 kDa and 7 kDa
-
-
?
VWF115 + H2O
?
show the reaction diagram
VWF115 D1614A mutant + H2O
10 kDa VWF115 fragment + 7 kDa VWF115 fragment
show the reaction diagram
-
Asp1614 VWFA2 domain fragment, spanning von Willebrand factor residues 1554-1668, generation of 2 cleavage products of 10 kDa and 7 kDa
-
-
?
VWF73 peptide + H2O
?
show the reaction diagram
-
von Willebrand factor-derived peptide substrate
-
-
?
VWF73 region of von Willebrand factor + H2O
?
show the reaction diagram
-
with this minimal substrate urea is not required for cleavage, minimal substrate for ADAMTS-13
-
?
VWFA2 peptide + H2O
?
show the reaction diagram
-
substrate based on a 78-amino acid sequence corresponding to the sequence Leu1591-Arg1668 of the von Willebrand factor A2 domain
-
-
?
VWFA2 peptide + H2O
VWFA2 peptide fragments
show the reaction diagram
-
A2 domain fragment of von Willebrand factor, cleavage of oxidized or nonoxidized VWFA2 peptide by ADAMTS13, cleavage of the Tyr1605-Met(O)1606 peptide bond by ADAMTS13, overview
-
-
?
additional information
?
-
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
proteins + H2O
peptides
show the reaction diagram
von Willebrand factor + H2O
2 peptides
show the reaction diagram
-
rapid physiological process that occurs on endothelial surfaces, reduced activity results in thrombotic thrombocytopenic pupura
-
?
von Willebrand factor + H2O
2 peptides of 140 kD and 65 kD
show the reaction diagram
-
cleavage of peptide bond Tyr842-Met843
-
?
von Willebrand factor + H2O
2 peptides of 140 kDa and 176 kDa
show the reaction diagram
von Willebrand factor + H2O
2 peptides of 176 kD and 140 kD
show the reaction diagram
-
rapid physiological process that occurs on endothelial surfaces, activity is reduced by 87-100% in patients with thrombotic thrombocytopenic pupura
-
?
von Willebrand factor + H2O
?
show the reaction diagram
von Willebrand factor + H2O
von Willebrand factor 140-kD fragment + von Willebrand factor 176-kD fragment
show the reaction diagram
von Willebrand factor + H2O
von Willebrand factor fragments
show the reaction diagram
von Willebrand factor 115 + H2O
10000 Da fragment of von Willebrand factor 115 + 7000 Da fragment of von Willebrand factor 115
show the reaction diagram
-
von Willebrand factor is cleaved at the Tyr1605-Met1606 bond in the von Willebrand factor A2 domain
-
-
?
additional information
?
-
METALS and IONS
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
Cu2+
-
activates
guanidine
-
activates
Mn2+
-
activates
Ni2+
-
activates
Sr2+
-
activating
Urea
-
activates
additional information
INHIBITORS
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
1,10-phenanthroline
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activity can be restored by Ca2+
Ca2+
-
enzyme catalytic activity in blood plasma is tuned by varying buffer calcium, with lower divalent ion concentrations enhancing cleavage
citrate
-
weak
Cl-
-
Chloride ions inhibit von Willebrand factor hydrolysis by ADAMTS-13 of the A1-A2-A3 and A1-A2 domains in the presence of either urea or high shear stress, whereas this effect is either absent or negligible when using A2 and A2-A3 domains
CoCl2
-
an increasing concentration of CoCl2 inhibits ADAMTS13 activity
H2O2
-
treatment with 25 nM myeloperoxidase plus 50 microM H2O2 reduces ADAMTS13 activity by >85%. Residues Met249, Met331, and Met496 in important functional domains of ADAMTS13 are oxidized to methionine sulfoxide in an HOCl concentration-dependent manner
heparin
-
inhibits ADAMTS13 activity through inhibition of enzyme binding to endothelial cell surfaces, overview
HOCl
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HOCl can oxidize methionine to methionine sulfoxide and tyrosine to chlorotyrosine, oxidation of VWF A2 peptide, at Met1606 and Tyr1605, markedly impairs ADAMTS13 cleavage. Oxidative modification by myeloperoxidase/H2O2 is similar to that produced by HOCl
Mn2+
-
ADAMTS13 activity is decreased in the presence of 0.9 mM Ni2+
MnSO4
-
an increasing concentration of MnSO4 inhibits ADAMTS13 activity
N-ethylmaleimide
-
slow and weak inhibition
Ni2+
-
ADAMTS13 activity is markedly decreased in the presence of 0.9 mM Ni2+
NiSO4
-
an increasing concentration of NiSO4 inhibits ADAMTS13 activity
peroxynitrite
-
formation of methionine sulfoxide by peroxynitrite at position 1606 of von Willebrand factor inhibits its cleavage by ADAMTS-13 a prothrombotic mechanism in diseases associated with oxidative stress, overview. Oxidation by peroxynitrite of purified VWF multimers inhibits ADAMTS-13 hydrolysis, but does not alter their electrophoretic pattern nor their ability to induce platelet agglutination by ristocetin. In vitro treatment of ADAMTS-13 with peroxynitrite over a concentration ranging from 0.050 to 0.250 mM causes a complete inhibition of the protease activity of the enzyme
plasma from patients with thrombotic thrombocytopenic purpurea
-
-
-
Pro-1645-Lys-1668 fragment of von Willebrand factor 73
-
-
-
Trypsin
-
inhibits ADAMTS13 activity through inhibition of enzyme binding to endothelial cell surfaces, overview
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VWFA2 domain
-
a C-terminal 32 kDa fragment of VWF, soluble VWFA2 domain effectively inhibits the binding of ADAMTS13 to immobilized VWFA2, and completely inhibits the interaction between ADAMTS13 and immobilized 64 kDa VWFA1A2A3 fragment
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W688X6-1
-
the monoclonal antibody W688X6-1 shows dose-dependent inhibitory activity toward ADAMTS13-mediated hydrolysis
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WH2-22-1A
-
the monoclonal antibody WH2-22-1A shows dose-dependent inhibitory activity toward ADAMTS13-mediated hydrolysis
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Z-Phe-Phe-CHN2
-
best peptidyl diazomethyl ketone inhibitor
additional information
-
ACTIVATING COMPOUND
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
Ristocetin
-
activates
Rituximab
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a monoclonal anti-CD 20 antibody, Rituximab leads to a prompt reduction in IgG antibody levels, followed by an increase in ADAMTS 13 activity increases. However, in patients receiving Rituximab electively, normalisation of ADAMTS 13 enzyme activity may be delayed for up to 3 months
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von Willebrand factor
-
binding to substrate von Willebrand factor D4 allosterically activates ADAMTS13, increasing its catalytic activity. Binding also positions ADAMTS13 on the substrate where it can act rapidly when shear stress exposes the scissile bond in the adjacent A2 domain
-
additional information
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KM VALUE [mM]
SUBSTRATE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
0.00662
DREQAPNLVYMVTGNPASDEIKRLPGDIQVVPIGVGPNANVQELERIGWPNAPILIQDFETLPREAPDLVLQRA
-
pH 7.5, 25°C, recombinant enzyme
0.00129 - 0.00432
FRET-VWF115 peptide
-
0.0032
FRETS-von Willebrand factor 73
-
apparent value, pH 7.4
-
0.0058
FRETSVWF73 peptide
-
pH 7.5, 37°C, recombinant ADAMTS13
-
0.00025
HRPH-A2-B
-
-
-
0.00065 - 0.00161
von Willebrand factor 115
-
0.00086 - 0.59
VWF115
-
0.37 - 0.47
VWF115 D1614A mutant
-
0.0017 - 0.022
VWF73 peptide
-
additional information
additional information
-
TURNOVER NUMBER [1/s]
SUBSTRATE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
0.72
DREQAPNLVYMVTGNPASDEIKRLPGDIQVVPIGVGPNANVQELERIGWPNAPILIQDFETLPREAPDLVLQRA
Homo sapiens
-
pH 7.5, 25°C, recombinant enzyme
0.31 - 1.31
FRET-VWF115 peptide
-
4.43
FRETSVWF73 peptide
Homo sapiens
-
pH 7.5, 37°C, recombinant ADAMTS13
-
0.67
HRPH-A2-B
Homo sapiens
-
-
-
0.14 - 0.52
von Willebrand factor 115
-
0.0019 - 0.71
VWF115
-
0.008 - 0.00875
VWF115 D1614A mutant
-
0.76 - 2.05
VWF73 peptide
-
additional information
additional information
Homo sapiens
-
kcat: 0.83 per min for FRETS-VWF73, a fluorescent synthetic peptide, corresponding to residues Asp1596-Arg1668 of von Willebrand factor domain A2 and containing the Tyr1605-Met1606 bond cleaved by ADAMTS13
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kcat/KM VALUE [1/mMs-1]
SUBSTRATE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
71.76 - 1016
FRET-VWF115 peptide
19392
763.8
FRETSVWF73 peptide
Homo sapiens
-
pH 7.5, 37°C, recombinant ADAMTS13
41342
821
von Willebrand factor 115
Homo sapiens
-
wild type enzyme, in 20 mM Tris-HCl (pH 7.8), 150 mM NaCl, 5 mM CaCl2, at 37°C
18521
15 - 830
VWF115
15127
50 - 57
VWF115 D1614A mutant
19391
74 - 733
VWF73 peptide
4447
Ki VALUE [mM]
INHIBITOR
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
0.012
Pro-1645-Lys-1668 fragment of von Willebrand factor 73
-
-
-
0.000028
VWFA2 domain
-
pH 7.8, 37°C
-
additional information
additional information
-
C-terminal fragments competitive inhibition binding, kinetics, overview
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IC50 VALUE [mM]
INHIBITOR
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
0.000072
VWFA2 domain
Homo sapiens
-
pH 7.8, 37°C
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SPECIFIC ACTIVITY [µmol/min/mg]
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
additional information
pH OPTIMUM
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
6
-
peptide substrates
7.5 - 8
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multimeric substrate
7.5
-
assay at
7.5 - 8
-
assay at
8 - 9
-
-
8.15
-
assay at
9 - 10
-
in presence of Ca2+
additional information
-
no activity at neutral pH at low salt concentration
TEMPERATURE OPTIMUM
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
22
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room temperature, assay at
25
-
assay at
pI VALUE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
5.1 - 5.5
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purified blood plasma ADAMTS13, isoelectric focusing
SOURCE TISSUE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
SOURCE
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a renal carcinoma cell line
Manually annotated by BRENDA team
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in cerebral corpora amylacea, that occur in aging brains and in patients with neurodegenerative conditions
Manually annotated by BRENDA team
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in astrocytes of aging brains and patients with neurodegenerative conditions. Corporae amylaceae include glycosylated material, ubiquitin, and an assortment of proteins derived from neuronal cytoplasm. Many of these proteins are not specifically localized to neurons or astrocytes, some components of corpora amylacea, such as complement proteins, are most abundantly expressed outside the central nervous system. Corporae amylaceae can result from a conglomeration of interacting proteins from degenerating neurons and from extravasated blood elements released after transient breakdown of the blood-brain barrier
Manually annotated by BRENDA team
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primary renal cortical cell cultures, real-time PCR and immunohistochemic analysis, overview
Manually annotated by BRENDA team
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ADAMTS13 and von Willebrand factor contents, overview
Manually annotated by BRENDA team
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ADAMTS13 and von Willebrand factor contents, overview
Manually annotated by BRENDA team
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enzyme expression
Manually annotated by BRENDA team
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thrombospondin1 and ADAMTS13 form complexes together in cells and in direct protein binding assays
Manually annotated by BRENDA team
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secretes the enzyme
Manually annotated by BRENDA team
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in both proximal and distal in healthy persons and in patients with renal disorders, e.g. tubulopathy
Manually annotated by BRENDA team
-
enzyme expression
Manually annotated by BRENDA team
-
at the surface of endothelial cells
Manually annotated by BRENDA team
-
only in urine from patients with renal disorder tubulopathy
Manually annotated by BRENDA team
additional information
LOCALIZATION
ORGANISM
UNIPROT
COMMENTARY hide
GeneOntology No.
LITERATURE
SOURCE
-
homogeneous distribution of wild-type ADAMTS13 in cis-Golgi and endoplasmic reticulum compartments. Reduction of ADAMTS13(Val88Met) in both compartments. ADAMTS13(Gly1239Val) fails to reach the cis-Golgi compartment and remains in the endoplasmic reticulum
Manually annotated by BRENDA team
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homogeneous distribution of wild-type ADAMTS13 in cis-Golgi and endoplasmic reticulum compartments. Reduction of ADAMTS13(Val88Met) in both compartments. ADAMTS13(Gly1239Val) fails to reach the cis-Golgi compartment and remains in the endoplasmic reticulum
Manually annotated by BRENDA team
MOLECULAR WEIGHT
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
45000
-
x * 45000, recombinant His-tagged ADAMTS13 ancillary domains, ADAMTS13-DTCS, SDS-PAGE
150000 - 190000
-
gel filtration
150000
170000
180000
-
x * 180000, SDS-PAGE
190000
195000
-
SDS-PAGE
300000
-
gel filtration
SUBUNITS
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
monomer
additional information
POSTTRANSLATIONAL MODIFICATION
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
glycoprotein
proteolytic modification
Crystallization/COMMENTARY
ORGANISM
UNIPROT
LITERATURE
in electron microscopy, wild-type ADAMTS13 adopts a closed conformation, gain-of-function mutant R568K/F592Y/R660K/Y661F/Y665F shows a more open conformation
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purified recombinant detagged ADAMTS13-DTCS, residues 287-685, an non-catalytic von Willebrand factor-exosite-containing ADAMTS13 fragment, sitting drop vapor diffusion method, mixing of 500 nl of protein solution and 500 nl reservoir solution containing 26% w/v PEG1500, 100 mM MES, pH 6.0, supplemented with 200 nl of 40% w/w pentaerythritol ethoxylate, equilibration for several days at 20°C. Os-derivative crystals by soaking native crystals in reservoir solution supplemented with 1 mM OsCl3 and 20% glycerol for several hours, X-ray diffraction structure determination and analysis at 2.6 A and 2.8 A resolution resulting in two crystal structures
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recombinant His-tagged ADAMTS13 ancillary domains, residues 287-685, sitting-drop vapour-diffusion method, mixing of 100 nl protein solution, containing about 20 mg/ml protein, with an equal amount of reservoir solution, containing 26% w/v PEG 1500, 100 mM MES, pH 6.0, equilibration against 0.1 ml of reservoir solution, at 20°C, for 24 h, crystals are soaked in a solution containing 20% glycerol, 26% PEG 1500, 100 mM MES, pH 6.0, for cryoprotection, X-ray diffraction structure determination and analysis at 2.6-2.8 A resolution, labeling with heavy-atom derivatives
-
pH STABILITY
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
6.1 - 8.8
-
the activity of purified pADAMTS13 is stable and relatively constant at pH ranges between pH 6.1 and 8.8 for 18 h storage at 4°C. However, ADAMTS13 activity is almost totally lost below pH 3.1
719761
TEMPERATURE STABILITY
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
22
-
stable for several days during purification at room temperature
GENERAL STABILITY
ORGANISM
UNIPROT
LITERATURE
ADAMTS-13 is highly stable in plasma that is stored frozen at -80°C
-
Purification/COMMENTARY
ORGANISM
UNIPROT
LITERATURE
A10-agarose column chromatography, HiTrap DEAE column chromatography, and Superdex 200 gel filtration
-
about 10000fold
-
from commercial factor VIII/vWF concentrate
-
HiLoad Superdex 200 gel filtration
-
Ni-NTA agarose column chromatography
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Ni-NTA column chromatography
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recombinant ADAMTS-13 from HEK293 cells by Zn2+-agarose affinity and anion exchange chromatography to homogeneity
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recombinant His-tagged ADAMTS13 ancillary domains, ADAMTS13-DTCS, consisting of residues 287-685, from CHO Lec cells by nickel affinity and cation exchange chromatography
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recombinant His-tagged ADAMTS13 by nickel affinity chromatography
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recombinant His-tagged ADAMTS13 from HEK-293 cells by anion exchange chromatography, Ni affinity chromatography, and gel filtration
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recombinant structure-based mutants of ADAMTS13-MDTCS residues 75-685 fragment with a C-terminal tobacco etch virus proteinase cleavage site followed by tandem His-tag sequences from CHO Lec 3.2.8.1 cells by nickel affinity chromatography and gel filtration
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recombinant tagged deletion mutants
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recombinant wild-type ADAMTS13 with a C-terminal Myc/His tag from HEK293 cells, and recombinant His-tagged truncated DAMTS13 mutants MP-Dis and MP by nickel affinity chromatography and gel filtration
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recombinant wild-type and mutant ADAMTS13 from HEK-293 cells by nickel affinity chromatography
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recombinant wild-type and mutant ADAMTS13s with a C-terminal Myc/His tag expressed from HEK-293 cells by nickel affinity chromatography
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Sephadex 26/10 gel filtration
-
to homogeneity
-
Cloned/COMMENTARY
ORGANISM
UNIPROT
LITERATURE
ADAMTS13 DNA and amino acid sequence determination and analysis of wild-type and mutant enzymes, genotyping and expression analysis in HEK-293 cells, overview
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ADAMTS13 genotyping, overview
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construction of ADAMTS13 epitope library
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DNA and amino acid sequence determination and analysis, genotyping
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DNA sequence determination and analysis
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DNA sequence determination and analysis, transient expression of wild-type and mutants as FLAG-tagged enzymes in HeLa cells
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DNA sequence determination of natural genetic variants, heterozygous polymorphism at Tyr/Cys1584 and other, overview
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expressed in CHO cells
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expressed in HEK-293 cells
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expressed in HeLa cells
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expressed in T-REx 293 cells
-
expression in HEK-293 cell
expression of a series of partial deletions in the A2 domain flanked with N- and C-terminal tags in Escherichia coli
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expression of ADAMTS-13 in HEK293 cells
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expression of ADAMTS13 in HEK-293 cells
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expression of FLAG-tagged ADAMTS13 in HEK-293 cells, co-expression with thrombospondin1
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expression of His-tagged ADAMTS13
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expression of structure-based mutants of ADAMTS13-MDTCS residues 75-685 fragment with a C-terminal tobacco etch virus proteinase cleavage site followed by tandem His-tag sequences in CHO Lec 3.2.8.1 cells. Transient expression of His-tagged mutant ADAMTS13-MDTCS domains using a cytomegalovirus promoter-driven expression vector in HeLa cells
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expression of the murine Adamts-13 cDNA in HEK 293 cells
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expression of wild-type ADAMTS13 with a C-terminal Myc/His tag in HEK293 cells, transient expression of His-tagged truncated DAMTS13 mutants MP-Dis and MP and point mutants in HEK-293T cells
-
expression of wild-type and mutant ADAMTS13 in HEK-293 cells
-
expression of wild-type and mutant ADAMTS13s with a C-terminal Myc/His tag expressed in HEK-293 cells
-
expression of wild-type and mutant enzyme forms in HEK-293 cells and CHO cells
-
expression of wild-type and mutant enzymes in HEK 293 and COS-7 cells
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gene Adamts13, DNA and amino acid sequence determination and analysis, expression of truncated ADAMTS13 in 129/Sv transgenic mice
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gene Adamts13, DNA and amino acid sequence determination, construction of self-inactivated lentiviral vector, and expression of functional His-tagged ADAMTS13 cDNA in COS-7 cells
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gene ADAMTS13, location of chromosome 9q34
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gene ADMTS13 and mutant variant truncated after the spacer domain, MDTCS, expression in transgenic mice
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stable expression in HEK-293 cells
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stable expression of His-tagged ADAMTS13 ancillary domains, ADAMTS13-DTCS, consisting of residues 287-685, in CHO Lec cells
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stable expression of His-tagged ADAMTS13 in HEK-293 cells
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transient expression in HEK293 cells
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transient expression in HeLa cells, in HepG2 cells, in RFL6 cells, and in COS-1 cells, highest expression level is obtained in HeLa cells
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transient expression of C-terminally tagged enzyme in COS-7 cells or expression in a Sf9 cells/baculovirus expression system, tagged with His6 or V5 epitopes
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transient expression of wild-type and mutants in HeLa cells as FLAG-tagged proteins
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ENGINEERING
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
A1033T
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naturally occuring mutation of ADAMTS13
A250V
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naturally occuring mutation of ADAMTS13
A596V
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naturally occuring mutation of ADAMTS13
A606P
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naturally occuring mutation of ADAMTS13
A732V
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naturally occuring mutation of ADAMTS13
C1024G
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naturally occuring mutation of ADAMTS13
C1213Y
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naturally occuring mutation of ADAMTS13
C311Y
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naturally occuring mutation of ADAMTS13
C347S
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naturally occuring mutation of ADAMTS13
C758R
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naturally occuring mutation of ADAMTS13
C908S
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naturally occuring mutation of ADAMTS13
C908Y
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naturally occuring mutation of ADAMTS13
C951G
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naturally occuring mutation of ADAMTS13
C977W
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deletion of 6 nucleotides GTGCCC at position 2930-2935, i.e. c.2930_2935del GTGCCC, in exon 23, leading to the replacement of Cys977 residue by a Trp
D187H
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mutation identified in a patient with pregnancy-onset thrombotic thrombocytopenic purpura. Mutation is located in the high affinity Ca2+-binding site in the metalloprotease domain of ADAMTS13. The homozygous mutation down-regulates ADAMTS13 activity in vitro. Impaired proteolytic activity is linked to unstable Ca2+ binding. In addition, the D187H mutation affects protein secretion in vitro
D235H
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naturally occuring mutation of ADAMTS13
D252N
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the ADAMTS13 VR2 single-point mutant is secreted as the wild type enzyme
D330A
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site-directed mutagenesis, the mutant shows activity similar to the wild-type enzyme
D340A
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site-directed mutagenesis, the mutant shows slightly reduced activity compared to the wild-type enzyme
D343A
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site-directed mutagenesis, the mutant shows slightly reduced activity compared to the wild-type enzyme
D500E
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point mutation in the RGD cysteine-rich domain, unaltered activity compared to the wild-type enzyme
delQ1624-R1641
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mutation minimally affects the rate of cleavage
E184A/L185A/D187A/R190A
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the proteolytic function of the mutant is severely affected
E184A/L185A/D187A/R190A/Q191A/V192N/R193A
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the mutant has abolished activity against von Willebrand factor 115
E212A
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site-directed mutagenesis of a Ca2+ binding site residue, the kinetic dissociation constant of ADAMTS13 for Ca2+ is dramatically reduced compared to the wild-type enzyme, Vmax of the mutant is also reduced by 75% compared to the wild-type
E627X
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naturally occuring mutation of ADAMTS13
E634K
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naturally occuring mutation of ADAMTS13
E740K
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naturally occuring mutation of ADAMTS13
G1239V
G525D
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naturally occuring mutation of ADAMTS13
G982R
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naturally occuring mutation of ADAMTS13
H234Q
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naturally occuring mutation of ADAMTS13
H96D
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naturally occuring mutation of ADAMTS13
I1217T
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naturally occuring mutation of ADAMTS13
I178T
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naturally occuring mutation of ADAMTS13
I673F
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naturally occuring mutation of ADAMTS13
I79M
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naturally occuring mutation of ADAMTS13
L185A
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the mutant exhibits wild type activity
L218A
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the ADAMTS13 VR2 single-point mutant is secreted poorly
L232Q
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naturally occuring mutation of ADAMTS13
L350G
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site-directed mutagenesis, the mutant shows highly reduced activity compared to the wild-type enzyme
L351G
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site-directed mutagenesis, the mutant shows slightly reduced activity compared to the wild-type enzyme
N146Q
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decreased secretion and von Willebrand factor cleaving activity
N552Q
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decreased secretion
N828Q
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decreased secretion and von Willebrand factor cleaving activity
P353L
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naturally occuring mutation of ADAMTS13
P457L
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naturally occuring mutation of ADAMTS13
P618A/A732V
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mutation induces secretion deficiency
P671L
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naturally occuring mutation of ADAMTS13
Q1302X
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naturally occuring mutation of ADAMTS13
Q191A
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the mutant exhibits wild type activity
Q197A
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the mutant exhibits wild type activity
Q333A
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site-directed mutagenesis, the mutant shows activity similar to the wild-type enzyme
Q449X
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naturally occuring mutation of ADAMTS13
Q44X
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naturally occuring mutation of ADAMTS13
Q456H
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naturally occuring mutation of ADAMTS13
Q929X
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naturally occuring mutation of ADAMTS13
R102C
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naturally occuring mutation of ADAMTS13
R1034X
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naturally occuring mutation of ADAMTS13
R1060W
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naturally occuring mutation of ADAMTS13
R1096H
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naturally occuring mutation of ADAMTS13
R1123C
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naturally occuring mutation of ADAMTS13
R1206X
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naturally occuring mutation of ADAMTS13
R1219W
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naturally occuring mutation of ADAMTS13
R1336
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mutation induces secretion deficiency
R1336W
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naturally occuring mutation of ADAMTS13
R190A
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the mutant shows 2fold reduced catalytic efficiency against von Willebrand factor 115 compared to the wild type enzyme
R193A
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the mutant shows 4fold reduced catalytic efficiency against von Willebrand factor 115 compared to the wild type enzyme
R193W
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naturally occuring mutation of ADAMTS13
R257A
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the ADAMTS13 VR2 single-point mutant is secreted as the wild type enzyme
R349A
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site-directed mutagenesis, the mutant shows highly reduced activity compared to the wild-type enzyme, the mutant enzyme shows increased activity with the mutant D1614A von Willebrand factor115 substrate compared to the wild-type enzyme
R349C
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naturally occuring mutation of ADAMTS13
R398H
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naturally occuring mutation of ADAMTS13
R484K
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naturally occuring mutation of ADAMTS13
R507Q
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naturally occuring mutation of ADAMTS13
R528G
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naturally occuring mutation of ADAMTS13
R568K/F592Y/R660K/Y661F/Y665F
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gain-of-function ADAMTS13 spacer domain variant, about 2.5fold more active than wild-type ADAMTS13, but cannot be further activated by anti-CUB monoclonal antibody or von Willebrand factor D4CK and is unable to bind or to be inhibited by the CUB1-2 domains
R625H
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naturally occuring mutation of ADAMTS13
R660A/Y661A
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site-directed mutagenesis
R660A/Y661A/Y665A
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site-directed mutagenesis, the ADAMTS13 variant, i.e. ADAMTS13-RYY, shows a 12fold reduced catalytic efficiency arising from over 25fold reduced substrate binding
R660A/Y665A
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site-directed mutagenesis
R692C
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naturally occuring mutation of ADAMTS13
R910X
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naturally occuring mutation of ADAMTS13
S119A
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site-directed mutagensis, mutant S119A has properties similar to natural mutant S119F
S119F/Q448E
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naturally occuring mutation in the ADAMTS13 metalloprotease domain. The mutant is expressed normally, but shows markedly impaired secretion
S203P
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naturally occuring mutation of ADAMTS13
S251A
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the ADAMTS13 VR2 single-point mutant is secreted poorly
S263C
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naturally occuring mutation of ADAMTS13
S903L
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naturally occuring mutation of ADAMTS13
T1226I
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naturally occuring mutation of ADAMTS13
T196A
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the mutant exhibits wild type activity
T196I
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naturally occuring mutation of ADAMTS13
T260A
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the ADAMTS13 VR2 single-point mutant is secreted as the wild type enzyme
T339R
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naturally occuring mutation of ADAMTS13
V192N
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the mutant exhibits wild type activity
V352G
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site-directed mutagenesis, the mutant shows reduced activity compared to the wild-type enzyme
V604I
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naturally occuring mutation of ADAMTS13
V832M
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naturally occuring mutation of ADAMTS13
W1016X
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naturally occuring mutation of ADAMTS13
W1245X
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naturally occuring mutation of ADAMTS13
W390C
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naturally occuring mutation of ADAMTS13
W390X
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naturally occuring mutation of ADAMTS13
Y304C
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naturally occuring mutation of ADAMTS13
Y658A
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site-directed mutagenesis
Y661A/Y665A
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site-directed mutagenesis
Y665A
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site-directed mutagenesis
additional information