Cloned (Comment) | Organism |
---|---|
expression of GST-tagged wild-type and mutant enzymes in Escherichia coli strain DH5alpha | Rattus norvegicus |
Protein Variants | Comment | Organism |
---|---|---|
A607G | site-directed mutagenesis, the mutation does not affect the overall fold of the protein, the mutant shows altered substrate specificity and kinetics with fluorogenic peptide substrates compared to the wild-type enzyme | Rattus norvegicus |
Y605A | site-directed mutagenesis, the mutation does not affect the overall fold of the protein, TOP Y605A is inhibited less efficiently by JA-2, the mutant shows altered substrate specificity and kinetics with fluorogenic peptide substrates compared to the wild-type enzyme | Rattus norvegicus |
Y605F | site-directed mutagenesis, the mutation does not affect the overall fold of the protein, the mutant shows altered substrate specificity and kinetics with fluorogenic peptide substrates compared to the wild-type enzyme | Rattus norvegicus |
Inhibitors | Comment | Organism | Structure |
---|---|---|---|
JA-2 | i.e. N-[1-(R,S)-carboxy-3-phenylpropyl]Ala-Aib-Tyrp-aminobenzoate, the aromatic ring of Tyr605 was an important anchor for its interaction with wild-type TOP | Rattus norvegicus |
KM Value [mM] | KM Value Maximum [mM] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|
0.0004 | - |
Abz-GFSIFRQEDDnp | pH 7.4, 37°C, recombinant GST-tagged wild-type enzyme | Rattus norvegicus | |
0.0007 | - |
Abz-GFSFFRQEDDnp | pH 7.4, 37°C, recombinant GST-tagged wild-type enzyme | Rattus norvegicus | |
0.0008 | - |
Abz-GFSWFRQEDDnp | pH 7.4, 37°C, recombinant GST-tagged wild-type enzyme | Rattus norvegicus | |
0.0009 | - |
Abz-GFSyFRQEDDnp | pH 7.4, 37°C, recombinant GST-tagged wild-type enzyme | Rattus norvegicus | |
0.0011 | - |
Abz-GFSRFRQEDDnp | pH 7.4, 37°C, recombinant GST-tagged wild-type enzyme | Rattus norvegicus | |
0.0012 | - |
Abz-GFSAFRQEDDnp | pH 7.4, 37°C, recombinant GST-tagged wild-type enzyme | Rattus norvegicus | |
0.0012 | - |
Abz-GFSQFRQEDDnp | pH 7.4, 37°C, recombinant GST-tagged wild-type enzyme | Rattus norvegicus | |
0.0014 | - |
Abz-GFSIFRQEDDnp | pH 7.4, 37°C, recombinant GST-tagged mutant Y605F | Rattus norvegicus | |
0.0015 | - |
2-aminobenzoyl-GFSPFRQ-(N-2,4-dinitrophenyl)ethylenediamine | pH 7.4, 37°C, recombinant GST-tagged mutant Y605F | Rattus norvegicus | |
0.0015 | - |
Abz-GFSWFRQEDDnp | pH 7.4, 37°C, recombinant GST-tagged mutant Y605F | Rattus norvegicus | |
0.0016 | - |
Abz-GFSLFRQEDDnp | pH 7.4, 37°C, recombinant GST-tagged wild-type enzyme | Rattus norvegicus | |
0.0021 | - |
Abz-GFSEFRQEDDnp | pH 7.4, 37°C, recombinant GST-tagged mutant Y605F | Rattus norvegicus | |
0.0022 | - |
2-aminobenzoyl-GFSPFRQ-(N-2,4-dinitrophenyl)ethylenediamine | pH 7.4, 37°C, recombinant GST-tagged wild-type enzyme | Rattus norvegicus | |
0.0023 | - |
Abz-GFSyFRQEDDnp | pH 7.4, 37°C, recombinant GST-tagged mutant Y605F | Rattus norvegicus | |
0.003 | - |
Abz-GFSEFRQEDDnp | pH 7.4, 37°C, recombinant GST-tagged mutant Y605A | Rattus norvegicus | |
0.0032 | - |
Abz-GFSLFRQEDDnp | pH 7.4, 37°C, recombinant GST-tagged mutant Y605F | Rattus norvegicus | |
0.0033 | - |
Abz-GFSFFRQEDDnp | pH 7.4, 37°C, recombinant GST-tagged mutant Y605F | Rattus norvegicus | |
0.0037 | - |
Abz-GFSSFRQEDDnp | pH 7.4, 37°C, recombinant GST-tagged wild-type enzyme | Rattus norvegicus | |
0.0043 | - |
2-aminobenzoyl-GFSPFRQ-(N-2,4-dinitrophenyl)ethylenediamine | pH 7.4, 37°C, recombinant GST-tagged mutant Y605A | Rattus norvegicus | |
0.0044 | - |
Abz-GFSRFRQEDDnp | pH 7.4, 37°C, recombinant GST-tagged mutant Y605F | Rattus norvegicus | |
0.0047 | - |
Abz-GFSAFRQEDDnp | pH 7.4, 37°C, recombinant GST-tagged mutant Y605F | Rattus norvegicus | |
0.0047 | - |
Abz-GFSSFRQEDDnp | pH 7.4, 37°C, recombinant GST-tagged mutant Y605F | Rattus norvegicus | |
0.005 | - |
Abz-GFSIFRQEDDnp | pH 7.4, 37°C, recombinant GST-tagged mutant Y605A | Rattus norvegicus | |
0.0054 | - |
Abz-GFSQFRQEDDnp | pH 7.4, 37°C, recombinant GST-tagged mutant Y605F | Rattus norvegicus | |
0.0058 | - |
Abz-GFSQFRQEDDnp | pH 7.4, 37°C, recombinant GST-tagged mutant Y605A | Rattus norvegicus | |
0.0074 | - |
Abz-GFSWFRQEDDnp | pH 7.4, 37°C, recombinant GST-tagged mutant Y605A | Rattus norvegicus | |
0.0075 | - |
Abz-GFSEFRQEDDnp | pH 7.4, 37°C, recombinant GST-tagged wild-type enzyme | Rattus norvegicus | |
0.0076 | - |
Abz-GFSHFRQEDDnp | pH 7.4, 37°C, recombinant GST-tagged wild-type enzyme | Rattus norvegicus | |
0.0086 | - |
Abz-GFSHFRQEDDnp | pH 7.4, 37°C, recombinant GST-tagged mutant Y605F | Rattus norvegicus | |
0.0096 | - |
Abz-GFSHFRQEDDnp | pH 7.4, 37°C, recombinant GST-tagged mutant Y605A | Rattus norvegicus | |
0.0098 | - |
Abz-GFSAFRQEDDnp | pH 7.4, 37°C, recombinant GST-tagged mutant Y605A | Rattus norvegicus | |
0.01 | - |
Abz-GFSRFRQEDDnp | pH 7.4, 37°C, recombinant GST-tagged mutant Y605A | Rattus norvegicus | |
0.0102 | - |
Abz-GFSLFRQEDDnp | pH 7.4, 37°C, recombinant GST-tagged mutant Y605A | Rattus norvegicus | |
0.011 | - |
Abz-GFSyFRQEDDnp | pH 7.4, 37°C, recombinant GST-tagged mutant Y605A | Rattus norvegicus | |
0.016 | - |
Abz-GFSFFRQEDDnp | pH 7.4, 37°C, recombinant GST-tagged mutant Y605A | Rattus norvegicus | |
0.02 | - |
Abz-GFSSFRQEDDnp | pH 7.4, 37°C, recombinant GST-tagged mutant Y605A | Rattus norvegicus |
Metals/Ions | Comment | Organism | Structure |
---|---|---|---|
Zn2+ | a zinc-dependent metallopeptidase | Rattus norvegicus |
Organism | UniProt | Comment | Textmining |
---|---|---|---|
Rattus norvegicus | - |
- |
- |
Purification (Comment) | Organism |
---|---|
recombinant GST-tagged wild-type and mutant enzymes from Escherichia coli strain DH5alpha by glutathione affinity chromatography to homogeneity | Rattus norvegicus |
Specific Activity Minimum [µmol/min/mg] | Specific Activity Maximum [µmol/min/mg] | Comment | Organism |
---|---|---|---|
additional information | - |
- |
Rattus norvegicus |
Substrates | Comment Substrates | Organism | Products | Comment (Products) | Rev. | Reac. |
---|---|---|---|---|---|---|
Abz-GFSAFRQ-EDDnp + H2O | - |
Rattus norvegicus | Abz-GFSA + FRQ-EDDnp | - |
? | |
Abz-GFSEFRQ-EDDnp + H2O | - |
Rattus norvegicus | Abz-GFSE + FRQ-EDDnp | - |
? | |
Abz-GFSFFRQ-EDDnp + H2O | - |
Rattus norvegicus | Abz-GFSF + FRQ-EDDnp | - |
? | |
Abz-GFSHFRQ-EDDnp + H2O | - |
Rattus norvegicus | Abz-GFSH + FRQ-EDDnp | - |
? | |
Abz-GFSIFRQ-EDDnp + H2O | - |
Rattus norvegicus | Abz-GFSI + FRQ-EDDnp | - |
? | |
Abz-GFSLFRQ-EDDnp + H2O | - |
Rattus norvegicus | Abz-GFSL + FRQ-EDDnp | - |
? | |
Abz-GFSPFRQ-EDDnp + H2O | - |
Rattus norvegicus | Abz-GFSP + FRQ-EDDnp | - |
? | |
Abz-GFSQFRQ-EDDnp + H2O | - |
Rattus norvegicus | Abz-GFSQ + FRQ-EDDnp | - |
? | |
Abz-GFSRFRQ-EDDnp + H2O | - |
Rattus norvegicus | Abz-GFSR + FRQ-EDDnp | - |
? | |
Abz-GFSSFRQ-EDDnp + H2O | - |
Rattus norvegicus | Abz-GFSS + FRQ-EDDnp | - |
? | |
Abz-GFSWFRQ-EDDnp + H2O | - |
Rattus norvegicus | Abz-GFSW + FRQ-EDDnp | - |
? | |
Abz-GFSYFRQ-EDDnp + H2O | - |
Rattus norvegicus | Abz-GFSY + FRQ-EDDnp | - |
? | |
additional information | cleavage sites of wild-type and mutant enzymes, overview | Rattus norvegicus | ? | - |
? |
Subunits | Comment | Organism |
---|---|---|
More | Ala607 of TOP contributes to the flexibility of the loops formed by residues 599-611, i.e. GHLAGGYDAQYYG, in TOP, overview | Rattus norvegicus |
Synonyms | Comment | Organism |
---|---|---|
More | the enzyme belongs to the zinc-dependent peptidases of the metallopeptidase M3 family | Rattus norvegicus |
TOP | - |
Rattus norvegicus |
Temperature Optimum [°C] | Temperature Optimum Maximum [°C] | Comment | Organism |
---|---|---|---|
37 | - |
assay at | Rattus norvegicus |
Turnover Number Minimum [1/s] | Turnover Number Maximum [1/s] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|
0.001 | - |
Abz-GFSIFRQEDDnp | pH 7.4, 37°C, recombinant GST-tagged wild-type enzyme | Rattus norvegicus | |
0.2 | - |
Abz-GFSWFRQEDDnp | pH 7.4, 37°C, recombinant GST-tagged mutant Y605A | Rattus norvegicus | |
0.6 | - |
Abz-GFSRFRQEDDnp | pH 7.4, 37°C, recombinant GST-tagged mutant Y605A | Rattus norvegicus | |
1.4 | - |
2-aminobenzoyl-GFSPFRQ-(N-2,4-dinitrophenyl)ethylenediamine | pH 7.4, 37°C, recombinant GST-tagged mutant Y605F | Rattus norvegicus | |
1.5 | - |
Abz-GFSSFRQEDDnp | pH 7.4, 37°C, recombinant GST-tagged mutant Y605A | Rattus norvegicus | |
1.5 | - |
Abz-GFSRFRQEDDnp | pH 7.4, 37°C, recombinant GST-tagged mutant Y605F | Rattus norvegicus | |
1.5 | - |
Abz-GFSEFRQEDDnp | pH 7.4, 37°C, recombinant GST-tagged wild-type enzyme | Rattus norvegicus | |
1.7 | - |
Abz-GFSSFRQEDDnp | pH 7.4, 37°C, recombinant GST-tagged mutant Y605F | Rattus norvegicus | |
1.8 | - |
Abz-GFSHFRQEDDnp | pH 7.4, 37°C, recombinant GST-tagged mutant Y605A | Rattus norvegicus | |
2.6 | - |
Abz-GFSAFRQEDDnp | pH 7.4, 37°C, recombinant GST-tagged mutant Y605F | Rattus norvegicus | |
2.7 | - |
Abz-GFSWFRQEDDnp | pH 7.4, 37°C, recombinant GST-tagged wild-type enzyme | Rattus norvegicus | |
3.2 | - |
Abz-GFSEFRQEDDnp | pH 7.4, 37°C, recombinant GST-tagged mutant Y605F | Rattus norvegicus | |
3.4 | - |
Abz-GFSLFRQEDDnp | pH 7.4, 37°C, recombinant GST-tagged wild-type enzyme | Rattus norvegicus | |
4.7 | - |
2-aminobenzoyl-GFSPFRQ-(N-2,4-dinitrophenyl)ethylenediamine | pH 7.4, 37°C, recombinant GST-tagged mutant Y605A | Rattus norvegicus | |
5 | - |
Abz-GFSIFRQEDDnp | pH 7.4, 37°C, recombinant GST-tagged mutant Y605A | Rattus norvegicus | |
5.3 | - |
Abz-GFSEFRQEDDnp | pH 7.4, 37°C, recombinant GST-tagged mutant Y605A | Rattus norvegicus | |
5.8 | - |
Abz-GFSQFRQEDDnp | pH 7.4, 37°C, recombinant GST-tagged mutant Y605A | Rattus norvegicus | |
6.2 | - |
Abz-GFSyFRQEDDnp | pH 7.4, 37°C, recombinant GST-tagged wild-type enzyme | Rattus norvegicus | |
6.3 | - |
2-aminobenzoyl-GFSPFRQ-(N-2,4-dinitrophenyl)ethylenediamine | pH 7.4, 37°C, recombinant GST-tagged wild-type enzyme | Rattus norvegicus | |
6.3 | - |
Abz-GFSQFRQEDDnp | pH 7.4, 37°C, recombinant GST-tagged wild-type enzyme | Rattus norvegicus | |
6.4 | - |
Abz-GFSyFRQEDDnp | pH 7.4, 37°C, recombinant GST-tagged mutant Y605F | Rattus norvegicus | |
6.7 | - |
Abz-GFSFFRQEDDnp | pH 7.4, 37°C, recombinant GST-tagged wild-type enzyme | Rattus norvegicus | |
7 | - |
Abz-GFSLFRQEDDnp | pH 7.4, 37°C, recombinant GST-tagged mutant Y605A | Rattus norvegicus | |
7 | - |
Abz-GFSRFRQEDDnp | pH 7.4, 37°C, recombinant GST-tagged wild-type enzyme | Rattus norvegicus | |
8.8 | - |
Abz-GFSSFRQEDDnp | pH 7.4, 37°C, recombinant GST-tagged wild-type enzyme | Rattus norvegicus | |
9.1 | - |
Abz-GFSWFRQEDDnp | pH 7.4, 37°C, recombinant GST-tagged mutant Y605F | Rattus norvegicus | |
10.2 | - |
Abz-GFSAFRQEDDnp | pH 7.4, 37°C, recombinant GST-tagged wild-type enzyme | Rattus norvegicus | |
10.4 | - |
Abz-GFSAFRQEDDnp | pH 7.4, 37°C, recombinant GST-tagged mutant Y605A | Rattus norvegicus | |
12.1 | - |
Abz-GFSQFRQEDDnp | pH 7.4, 37°C, recombinant GST-tagged mutant Y605F | Rattus norvegicus | |
12.7 | - |
Abz-GFSFFRQEDDnp | pH 7.4, 37°C, recombinant GST-tagged mutant Y605A | Rattus norvegicus | |
13.6 | - |
Abz-GFSIFRQEDDnp | pH 7.4, 37°C, recombinant GST-tagged mutant Y605F | Rattus norvegicus | |
13.6 | - |
Abz-GFSLFRQEDDnp | pH 7.4, 37°C, recombinant GST-tagged mutant Y605F | Rattus norvegicus | |
15.1 | - |
Abz-GFSHFRQEDDnp | pH 7.4, 37°C, recombinant GST-tagged mutant Y605F | Rattus norvegicus | |
20.4 | - |
Abz-GFSyFRQEDDnp | pH 7.4, 37°C, recombinant GST-tagged mutant Y605A | Rattus norvegicus | |
24.7 | - |
Abz-GFSHFRQEDDnp | pH 7.4, 37°C, recombinant GST-tagged wild-type enzyme | Rattus norvegicus | |
27.3 | - |
Abz-GFSFFRQEDDnp | pH 7.4, 37°C, recombinant GST-tagged mutant Y605F | Rattus norvegicus |
pH Optimum Minimum | pH Optimum Maximum | Comment | Organism |
---|---|---|---|
7.4 | - |
assay at | Rattus norvegicus |
pH Minimum | pH Maximum | Comment | Organism |
---|---|---|---|
additional information | - |
pH-dependence of wild-type and mutant enzymes, overview | Rattus norvegicus |