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 | 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 | Comment | Organism | Structure |
---|---|---|---|
EDTA | - |
Homo sapiens |
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 | Comment | Organism | GeneOntology No. | Textmining |
---|---|---|---|---|
extracellular | - |
Homo sapiens | - |
- |
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 | 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 | UniProt | Comment | Textmining |
---|---|---|---|
Homo sapiens | - |
- |
- |
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 | Comment | Organism | Textmining |
---|---|---|---|
plasma | - |
Homo sapiens | - |
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 | Comment | Organism |
---|---|---|
ADAMTS13 | - |
Homo sapiens |
ADAMTS13 metalloprotease | - |
Homo sapiens |
Temperature Optimum [°C] | Temperature Optimum Maximum [°C] | Comment | Organism |
---|---|---|---|
37 | - |
assay at | Homo sapiens |
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 Minimum | pH Optimum Maximum | Comment | Organism |
---|---|---|---|
7.8 | - |
assay at | Homo sapiens |
General Information | Comment | Organism |
---|---|---|
physiological function | ADAMTS13 regulates the multimeric size of von Willebrand factor | Homo sapiens |
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 |