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Literature summary for 3.4.24.87 extracted from

  • Gardner, M.; Chion, C.; Groot, R.; Shah, A.; Crawley, J.; Lane, D.
    A functional calcium-binding site in the metalloprotease domain of ADAMTS13 (2009), Blood, 113, 1149-1157.
    View publication on PubMed
Search for more literature for 3.4.24.87

Cloned(Commentary)

Cloned (Comment) Organism
expression of wild-type and mutant ADAMTS13s with a C-terminal Myc/His tag expressed in HEK-293 cells Homo sapiens

Protein Variants

Protein Variants Comment Organism
D187A 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%, and Kcat/Km 13fold, compared to the wild-type Homo sapiens
E184A 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 Homo sapiens
E212A 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 Homo sapiens

Inhibitors

Inhibitors Comment Organism Structure
EDTA
-
 Homo sapiens

KM Value [mM]

KM Value [mM] KM Value Maximum [mM] Substrate Comment Organism Structure
additional information
-
 additional information Ca2+ binding kinetics of wild-type and mutant enzymes, overview Homo sapiens
0.00129
-
 FRET-VWF115 peptide pH 7.8, 37°C, recombinant wild-type enzyme Homo sapiens
0.00432
-
 FRET-VWF115 peptide pH 7.8, 37°C, recombinant mutant D187A Homo sapiens

Localization

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

Metals/Ions

Metals/Ions Comment Organism Structure
Ba2+ activates Homo sapiens
Ca2+ highly dependent on, Ca2+ induces a conformational change in ADAMTS13, but only under low-ionic-strength conditions, which is abolished in mutants E83A and D173A. The Ca2+-binding site in proximity to the ADAMTS13 active site comprises Glu184, Asp187, and Glu212, identified by homology modelling. Mutagenesis of these residues within this site to alanine dramatically attenuates the KD(app) for Ca2+ of ADAMTS13, and for D187A and E212A also reduces the Vmax to approximately 25% of wild-type. The major influence of Ca2+ on ADAMTS13 function is mediated through binding to a high affinity site adjacent to its active site cleft. The short sequence spanning residues 162 to 168 of the metalloprotease domain is not a Ca2+ binding site Homo sapiens
Cu2+ activates Homo sapiens
Mg2+ activates Homo sapiens
Mn2+ activates Homo sapiens
additional information Ca2+ is the most potent activating divalent cation out of Ca2+, Ba2+, Mg2+, Mn2+, Ni2+, and Cu2+ Homo sapiens
Ni2+ activates Homo sapiens
Zn2+ dependent on 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 specific cleavage of ultra-large von Willebrand factor multimers ?
-
 ?

Organism

Organism UniProt Comment Textmining
Homo sapiens
-
-
-

Purification (Commentary)

Purification (Comment) Organism
recombinant wild-type and mutant ADAMTS13s with a C-terminal Myc/His tag expressed from HEK-293 cells by nickel affinity chromatography 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.
FRET-VWF115 peptide + H2O von Willebrand factor-derived peptide substrate comprising residues 1554-1668 Homo sapiens ?
-
 ?
additional information molecular modeling of ADAMTS13 metalloprotease domain using its sequence homology to adamalysin II and the crystal structure Homo sapiens ?
-
 ?
von Willebrand factor + H2O specific cleavage of ultra-large von Willebrand factor multimers Homo sapiens ?
-
 ?

Synonyms

Synonyms Comment Organism
ADAMTS13
-
 Homo sapiens
ADAMTS13 metalloprotease
-
 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
0.31
-
 FRET-VWF115 peptide pH 7.8, 37°C, recombinant mutant D187A Homo sapiens
1.31
-
 FRET-VWF115 peptide pH 7.8, 37°C, recombinant wild-type enzyme Homo sapiens

pH Optimum

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

General Information

General Information Comment Organism
physiological function ADAMTS13 regulates the multimeric size of von Willebrand factor Homo sapiens

kcat/KM [mM/s]

kcat/KM Value [1/mMs-1] kcat/KM Value Maximum [1/mMs-1] Substrate Comment Organism Structure
71.76
-
 FRET-VWF115 peptide pH 7.8, 37°C, recombinant mutant D187A Homo sapiens
1016
-
 FRET-VWF115 peptide pH 7.8, 37°C, recombinant wild-type enzyme Homo sapiens