Any feedback?
Please rate this page
(literature.php)
(0/150)

BRENDA support

Literature summary for 3.4.24.15 extracted from

  • Machado, M.F.; Rioli, V.; Dalio, F.M.; Castro, L.M.; Juliano, M.A.; Tersariol, I.L.; Ferro, E.S.; Juliano, L.; Oliveira, V.
    The role of Tyr605 and Ala607 of thimet oligopeptidase and Tyr606 and Gly608 of neurolysin in substrate hydrolysis and inhibitor binding (2007), Biochem. J., 404, 279-288.
    View publication on PubMedView publication on EuropePMC

Cloned(Commentary)

Cloned (Comment) Organism
expression of GST-tagged wild-type and mutant enzymes in Escherichia coli strain DH5alpha Rattus norvegicus

Protein Variants

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

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 [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

Metals/Ions Comment Organism Structure
Zn2+ a zinc-dependent metallopeptidase Rattus norvegicus

Organism

Organism UniProt Comment Textmining
Rattus norvegicus
-
-
-

Purification (Commentary)

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 [micromol/min/mg]

Specific Activity Minimum [µmol/min/mg] Specific Activity Maximum [µmol/min/mg] Comment Organism
additional information
-
-
Rattus norvegicus

Substrates and Products (Substrate)

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

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

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 [°C] Temperature Optimum Maximum [°C] Comment Organism
37
-
assay at Rattus norvegicus

Turnover Number [1/s]

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

pH Optimum Minimum pH Optimum Maximum Comment Organism
7.4
-
assay at Rattus norvegicus

pH Range

pH Minimum pH Maximum Comment Organism
additional information
-
pH-dependence of wild-type and mutant enzymes, overview Rattus norvegicus