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Literature summary for 3.4.24.16 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
Search for more literature for 3.4.24.16

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
G608A 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
Y606A 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
Y606F 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
0.0006
-
 2-aminobenzoyl-GFSPFRQ-(N-2,4-dinitrophenyl)ethylenediamine pH 7.4, 37°C, recombinant GST-tagged wild-type enzyme Rattus norvegicus
0.0007
-
 2-aminobenzoyl-GFSPFRQ-(N-2,4-dinitrophenyl)ethylenediamine pH 7.4, 37°C, recombinant GST-tagged mutant Y606F Rattus norvegicus
0.0008
-
 Abz-GFSRFRQEDDnp pH 7.4, 37°C, recombinant GST-tagged mutant G608A Rattus norvegicus
0.0008
-
 Abz-GFSIFRQEDDnp pH 7.4, 37°C, recombinant GST-tagged mutant G608A Rattus norvegicus
0.0008
-
 Abz-GFSIFRQEDDnp 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-GFSAFRQEDDnp pH 7.4, 37°C, recombinant GST-tagged mutant G608A Rattus norvegicus
0.0011
-
 Abz-GFSAFRQEDDnp pH 7.4, 37°C, recombinant GST-tagged wild-type enzyme Rattus norvegicus
0.0012
-
 Abz-GFSyFRQEDDnp pH 7.4, 37°C, recombinant GST-tagged mutant G608A Rattus norvegicus
0.0013
-
 Abz-GFSRFRQEDDnp pH 7.4, 37°C, recombinant GST-tagged wild-type enzyme Rattus norvegicus
0.0015
-
 Abz-GFSFFRQEDDnp pH 7.4, 37°C, recombinant GST-tagged wild-type enzyme Rattus norvegicus
0.0016
-
 Abz-GFSQFRQEDDnp pH 7.4, 37°C, recombinant GST-tagged wild-type enzyme Rattus norvegicus
0.0018
-
 Abz-GFSLFRQEDDnp pH 7.4, 37°C, recombinant GST-tagged wild-type enzyme Rattus norvegicus
0.0018
-
 Abz-GFSHFRQEDDnp pH 7.4, 37°C, recombinant GST-tagged wild-type enzyme Rattus norvegicus
0.0019
-
 Abz-GFSLFRQEDDnp pH 7.4, 37°C, recombinant GST-tagged mutant G608A Rattus norvegicus
0.0019
-
 Abz-GFSRFRQEDDnp pH 7.4, 37°C, recombinant GST-tagged mutant Y606F Rattus norvegicus
0.0022
-
 Abz-GFSWFRQEDDnp pH 7.4, 37°C, recombinant GST-tagged mutant Y606A Rattus norvegicus
0.0024
-
 Abz-GFSEFRQEDDnp pH 7.4, 37°C, recombinant GST-tagged wild-type enzyme Rattus norvegicus
0.0025
-
 Abz-GFSFFRQEDDnp pH 7.4, 37°C, recombinant GST-tagged mutant G608A Rattus norvegicus
0.0026
-
 Abz-GFSRFRQEDDnp pH 7.4, 37°C, recombinant GST-tagged mutant Y606A Rattus norvegicus
0.0029
-
 Abz-GFSWFRQEDDnp pH 7.4, 37°C, recombinant GST-tagged mutant G608A Rattus norvegicus
0.0031
-
 2-aminobenzoyl-GFSPFRQ-(N-2,4-dinitrophenyl)ethylenediamine pH 7.4, 37°C, recombinant GST-tagged mutant Y606A Rattus norvegicus
0.0038
-
 Abz-GFSyFRQEDDnp pH 7.4, 37°C, recombinant GST-tagged mutant Y606F Rattus norvegicus
0.0048
-
 Abz-GFSIFRQEDDnp pH 7.4, 37°C, recombinant GST-tagged mutant Y606F Rattus norvegicus
0.0048
-
 Abz-GFSLFRQEDDnp pH 7.4, 37°C, recombinant GST-tagged mutant Y606F Rattus norvegicus
0.005
-
 Abz-GFSEFRQEDDnp pH 7.4, 37°C, recombinant GST-tagged mutant Y606F Rattus norvegicus
0.0053
-
 Abz-GFSEFRQEDDnp pH 7.4, 37°C, recombinant GST-tagged mutant Y606A Rattus norvegicus
0.0054
-
 Abz-GFSQFRQEDDnp pH 7.4, 37°C, recombinant GST-tagged mutant G608A Rattus norvegicus
0.0056
-
 Abz-GFSSFRQEDDnp pH 7.4, 37°C, recombinant GST-tagged mutant G608A Rattus norvegicus
0.0058
-
 Abz-GFSAFRQEDDnp pH 7.4, 37°C, recombinant GST-tagged mutant Y606F Rattus norvegicus
0.0059
-
 Abz-GFSHFRQEDDnp pH 7.4, 37°C, recombinant GST-tagged mutant G608A Rattus norvegicus
0.006
-
 Abz-GFSWFRQEDDnp pH 7.4, 37°C, recombinant GST-tagged mutant Y606F Rattus norvegicus
0.0061
-
 Abz-GFSLFRQEDDnp pH 7.4, 37°C, recombinant GST-tagged mutant Y606A Rattus norvegicus
0.0065
-
 Abz-GFSyFRQEDDnp pH 7.4, 37°C, recombinant GST-tagged mutant Y606A Rattus norvegicus
0.0074
-
 Abz-GFSSFRQEDDnp pH 7.4, 37°C, recombinant GST-tagged mutant Y606A Rattus norvegicus
0.0074
-
 Abz-GFSSFRQEDDnp pH 7.4, 37°C, recombinant GST-tagged mutant Y606F Rattus norvegicus
0.0078
-
 Abz-GFSAFRQEDDnp pH 7.4, 37°C, recombinant GST-tagged mutant Y606A Rattus norvegicus
0.0083
-
 Abz-GFSRFRQEDDnp pH 7.4, 37°C, recombinant GST-tagged mutant G608A Rattus norvegicus
0.0096
-
 Abz-GFSQFRQEDDnp pH 7.4, 37°C, recombinant GST-tagged mutant Y606F Rattus norvegicus
0.0096
-
 Abz-GFSyFRQEDDnp pH 7.4, 37°C, recombinant GST-tagged wild-type enzyme Rattus norvegicus
0.0098
-
 Abz-GFSFFRQEDDnp pH 7.4, 37°C, recombinant GST-tagged mutant Y606F Rattus norvegicus
0.0101
-
 Abz-GFSFFRQEDDnp pH 7.4, 37°C, recombinant GST-tagged mutant Y606A Rattus norvegicus
0.0104
-
 Abz-GFSHFRQEDDnp pH 7.4, 37°C, recombinant GST-tagged mutant Y606F Rattus norvegicus
0.0106
-
 Abz-GFSHFRQEDDnp pH 7.4, 37°C, recombinant GST-tagged mutant Y606A Rattus norvegicus
0.0111
-
 Abz-GFSSFRQEDDnp pH 7.4, 37°C, recombinant GST-tagged wild-type enzyme Rattus norvegicus
0.021
-
 Abz-GFSQFRQEDDnp pH 7.4, 37°C, recombinant GST-tagged mutant Y606A 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 Gly608 of NEL contributes to the flexibility of the loops formed by residues 600-612, i.e. GHLAGGYDGQYYG, in NEL, overview Rattus norvegicus

Synonyms

Synonyms Comment Organism
More the enzyme belongs to the zinc-dependent peptidases of the metallopeptidase M3 family Rattus norvegicus
NEL
-
 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
0.006
-
 2-aminobenzoyl-GFSPFRQ-(N-2,4-dinitrophenyl)ethylenediamine pH 7.4, 37°C, recombinant GST-tagged mutant Y606A Rattus norvegicus
0.01
-
 Abz-GFSIFRQEDDnp pH 7.4, 37°C, recombinant GST-tagged mutant Y606F Rattus norvegicus
0.04
-
 Abz-GFSIFRQEDDnp pH 7.4, 37°C, recombinant GST-tagged mutant G608A Rattus norvegicus
0.05
-
 Abz-GFSHFRQEDDnp pH 7.4, 37°C, recombinant GST-tagged mutant Y606A Rattus norvegicus
0.06
-
 Abz-GFSSFRQEDDnp pH 7.4, 37°C, recombinant GST-tagged mutant Y606A Rattus norvegicus
0.09
-
 Abz-GFSFFRQEDDnp pH 7.4, 37°C, recombinant GST-tagged mutant Y606A Rattus norvegicus
0.1
-
 Abz-GFSLFRQEDDnp pH 7.4, 37°C, recombinant GST-tagged mutant Y606A Rattus norvegicus
0.1
-
 Abz-GFSEFRQEDDnp pH 7.4, 37°C, recombinant GST-tagged mutant Y606A Rattus norvegicus
0.1
-
 2-aminobenzoyl-GFSPFRQ-(N-2,4-dinitrophenyl)ethylenediamine pH 7.4, 37°C, recombinant GST-tagged mutant Y606F Rattus norvegicus
0.2
-
 Abz-GFSAFRQEDDnp pH 7.4, 37°C, recombinant GST-tagged mutant G608A Rattus norvegicus
0.2
-
 Abz-GFSSFRQEDDnp pH 7.4, 37°C, recombinant GST-tagged mutant Y606F Rattus norvegicus
0.2
-
 Abz-GFSHFRQEDDnp pH 7.4, 37°C, recombinant GST-tagged mutant Y606F Rattus norvegicus
0.2
-
 Abz-GFSIFRQEDDnp pH 7.4, 37°C, recombinant GST-tagged wild-type enzyme Rattus norvegicus
0.3
-
 Abz-GFSyFRQEDDnp pH 7.4, 37°C, recombinant GST-tagged mutant Y606A Rattus norvegicus
0.3
-
 2-aminobenzoyl-GFSPFRQ-(N-2,4-dinitrophenyl)ethylenediamine pH 7.4, 37°C, recombinant GST-tagged wild-type enzyme Rattus norvegicus
0.3
-
 Abz-GFSWFRQEDDnp pH 7.4, 37°C, recombinant GST-tagged wild-type enzyme Rattus norvegicus
0.4
-
 Abz-GFSRFRQEDDnp pH 7.4, 37°C, recombinant GST-tagged mutant Y606A Rattus norvegicus
0.5
-
 Abz-GFSAFRQEDDnp pH 7.4, 37°C, recombinant GST-tagged mutant Y606A Rattus norvegicus
0.6
-
 Abz-GFSQFRQEDDnp pH 7.4, 37°C, recombinant GST-tagged mutant Y606F Rattus norvegicus
0.7
-
 Abz-GFSRFRQEDDnp pH 7.4, 37°C, recombinant GST-tagged mutant G608A Rattus norvegicus
0.7
-
 Abz-GFSQFRQEDDnp pH 7.4, 37°C, recombinant GST-tagged mutant Y606A Rattus norvegicus
0.7
-
 Abz-GFSLFRQEDDnp pH 7.4, 37°C, recombinant GST-tagged mutant Y606F Rattus norvegicus
0.7
-
 Abz-GFSEFRQEDDnp pH 7.4, 37°C, recombinant GST-tagged wild-type enzyme Rattus norvegicus
0.8
-
 Abz-GFSAFRQEDDnp pH 7.4, 37°C, recombinant GST-tagged mutant Y606F Rattus norvegicus
0.9
-
 Abz-GFSRFRQEDDnp pH 7.4, 37°C, recombinant GST-tagged mutant Y606F Rattus norvegicus
1
-
 Abz-GFSWFRQEDDnp pH 7.4, 37°C, recombinant GST-tagged mutant Y606A Rattus norvegicus
1
-
 Abz-GFSyFRQEDDnp pH 7.4, 37°C, recombinant GST-tagged wild-type enzyme Rattus norvegicus
1.1
-
 Abz-GFSFFRQEDDnp pH 7.4, 37°C, recombinant GST-tagged mutant Y606F Rattus norvegicus
1.2
-
 Abz-GFSEFRQEDDnp pH 7.4, 37°C, recombinant GST-tagged mutant Y606F Rattus norvegicus
1.3
-
 Abz-GFSyFRQEDDnp pH 7.4, 37°C, recombinant GST-tagged mutant Y606F Rattus norvegicus
1.5
-
 Abz-GFSWFRQEDDnp pH 7.4, 37°C, recombinant GST-tagged mutant Y606F Rattus norvegicus
2.4
-
 Abz-GFSLFRQEDDnp pH 7.4, 37°C, recombinant GST-tagged mutant G608A Rattus norvegicus
2.4
-
 Abz-GFSWFRQEDDnp pH 7.4, 37°C, recombinant GST-tagged mutant G608A Rattus norvegicus
2.4
-
 Abz-GFSSFRQEDDnp pH 7.4, 37°C, recombinant GST-tagged mutant G608A Rattus norvegicus
2.5
-
 Abz-GFSHFRQEDDnp pH 7.4, 37°C, recombinant GST-tagged wild-type enzyme Rattus norvegicus
2.6
-
 Abz-GFSAFRQEDDnp pH 7.4, 37°C, recombinant GST-tagged wild-type enzyme Rattus norvegicus
3.3
-
 Abz-GFSRFRQEDDnp pH 7.4, 37°C, recombinant GST-tagged mutant G608A Rattus norvegicus
3.4
-
 Abz-GFSFFRQEDDnp pH 7.4, 37°C, recombinant GST-tagged wild-type enzyme Rattus norvegicus
3.5
-
 Abz-GFSLFRQEDDnp pH 7.4, 37°C, recombinant GST-tagged wild-type enzyme Rattus norvegicus
3.6
-
 Abz-GFSQFRQEDDnp pH 7.4, 37°C, recombinant GST-tagged wild-type enzyme Rattus norvegicus
4.1
-
 Abz-GFSyFRQEDDnp pH 7.4, 37°C, recombinant GST-tagged mutant G608A Rattus norvegicus
4.8
-
 Abz-GFSRFRQEDDnp pH 7.4, 37°C, recombinant GST-tagged wild-type enzyme Rattus norvegicus
4.9
-
 Abz-GFSFFRQEDDnp pH 7.4, 37°C, recombinant GST-tagged mutant G608A Rattus norvegicus
5.9
-
 Abz-GFSQFRQEDDnp pH 7.4, 37°C, recombinant GST-tagged mutant G608A Rattus norvegicus
6.3
-
 Abz-GFSHFRQEDDnp pH 7.4, 37°C, recombinant GST-tagged mutant G608A Rattus norvegicus
7.5
-
 Abz-GFSSFRQEDDnp pH 7.4, 37°C, recombinant GST-tagged wild-type enzyme 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