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Literature summary for 3.4.24.87 extracted from

  • de Groot, R.; Bardhan, A.; Ramroop, N.; Lane, D.A.; Crawley, J.T.
    Essential role of the disintegrin-like domain in ADAMTS13 function (2009), Blood, 113, 5609-5616.
    View publication on PubMed

Cloned(Commentary)

Cloned (Comment) Organism
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 Homo sapiens

Protein Variants

Protein Variants Comment Organism
D330A site-directed mutagenesis, the mutant shows activity similar to the wild-type enzyme Homo sapiens
D340A site-directed mutagenesis, the mutant shows slightly reduced activity compared to the wild-type enzyme Homo sapiens
D343A site-directed mutagenesis, the mutant shows slightly reduced activity compared to the wild-type enzyme Homo sapiens
L350G site-directed mutagenesis, the mutant shows highly reduced activity compared to the wild-type enzyme Homo sapiens
L351G site-directed mutagenesis, the mutant shows slightly reduced activity compared to the wild-type enzyme Homo sapiens
additional information construction of truncated DAMTS13 mutants MP-Dis and MP, which have a molecular weight of 30 kDa and 40 kDa, respectively Homo sapiens
Q333A site-directed mutagenesis, the mutant shows activity similar to the wild-type enzyme Homo sapiens
R349A 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 Homo sapiens
V352G site-directed mutagenesis, the mutant shows reduced activity compared to the wild-type enzyme Homo sapiens

KM Value [mM]

KM Value [mM] KM Value Maximum [mM] Substrate Comment Organism Structure
additional information
-
additional information kinetic analysis of ADAMTS13 disintegrin-like domain mutants Homo sapiens
0.09
-
VWF115 pH 7.8, 37°C, recombinant mutant R349A Homo sapiens
0.37
-
VWF115 D1614A mutant pH 7.8, 37°C, recombinant wild-type enzyme Homo sapiens
0.47
-
VWF115 D1614A mutant pH 7.8, 37°C, recombinant mutant R349A Homo sapiens
0.59
-
VWF115 pH 7.8, 37°C, recombinant wild-type enzyme Homo sapiens

Localization

Localization Comment Organism GeneOntology No. Textmining
extracellular
-
Homo sapiens
-
-

Metals/Ions

Metals/Ions Comment Organism Structure
Ca2+ required Homo sapiens
Zn2+ required, metalloprotease Homo sapiens

Molecular Weight [Da]

Molecular Weight [Da] Molecular Weight Maximum [Da] Comment Organism
190000
-
x * 190000, recombinant wild-type enzyme, SDS-PAGE Homo sapiens

Natural Substrates/ Products (Substrates)

Natural Substrates Organism Comment (Nat. Sub.) Natural Products Comment (Nat. Pro.) Rev. Reac.
von Willebrand factor + H2O Homo sapiens
-
?
-
?

Organism

Organism UniProt Comment Textmining
Homo sapiens
-
-
-

Purification (Commentary)

Purification (Comment) Organism
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 Homo sapiens

Source Tissue

Source Tissue Comment Organism Textmining
plasma
-
Homo sapiens
-

Substrates and Products (Substrate)

Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
additional information role for ADAMTS13 disintegrin-like domains in substrate recognition and proteolysis, homology modeling, overview. Residues Arg349, Leu350, and Val352 are predicted to form a cluster on the surface of the ADAMTS13 disintegrin-like domain immediately adjacent to the active-site cleft Homo sapiens ?
-
?
von Willebrand factor + H2O
-
Homo sapiens ?
-
?
VWF115 + H2O VWFA2 domain fragment, spanning von Willebrand factor residues 1554-1668, generation of 2 cleavage products of 10 kDa and 7 kDa Homo sapiens 10 kDa VWF115 fragment + 7 kDa VWF115 fragment
-
?
VWF115 D1614A mutant + H2O Asp1614 VWFA2 domain fragment, spanning von Willebrand factor residues 1554-1668, generation of 2 cleavage products of 10 kDa and 7 kDa Homo sapiens 10 kDa VWF115 fragment + 7 kDa VWF115 fragment
-
?

Subunits

Subunits Comment Organism
? x * 190000, recombinant wild-type enzyme, SDS-PAGE Homo sapiens

Synonyms

Synonyms Comment Organism
ADAMTS13
-
Homo sapiens

Temperature Optimum [°C]

Temperature Optimum [°C] Temperature Optimum Maximum [°C] Comment Organism
37
-
assay at Homo sapiens

Turnover Number [1/s]

Turnover Number Minimum [1/s] Turnover Number Maximum [1/s] Substrate Comment Organism Structure
0.0019
-
VWF115 pH 7.8, 37°C, recombinant wild-type enzyme Homo sapiens
0.0062
-
VWF115 pH 7.8, 37°C, recombinant mutant R349A Homo sapiens
0.008
-
VWF115 D1614A mutant pH 7.8, 37°C, recombinant wild-type enzyme Homo sapiens
0.00875
-
VWF115 D1614A mutant pH 7.8, 37°C, recombinant mutant R349A Homo sapiens

pH Optimum

pH Optimum Minimum pH Optimum Maximum Comment Organism
7.8
-
assay at Homo sapiens

kcat/KM [mM/s]

kcat/KM Value [1/mMs-1] kcat/KM Value Maximum [1/mMs-1] Substrate Comment Organism Structure
15
-
VWF115 pH 7.8, 37°C, recombinant mutant R349A Homo sapiens
50
-
VWF115 D1614A mutant pH 7.8, 37°C, recombinant wild-type enzyme Homo sapiens
57
-
VWF115 D1614A mutant pH 7.8, 37°C, recombinant mutant R349A Homo sapiens
324
-
VWF115 pH 7.8, 37°C, recombinant wild-type enzyme Homo sapiens