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Literature summary for 3.4.24.15 extracted from

  • Lim, E.J.; Sampath, S.; Coll-Rodriguez, J.; Schmidt, J.; Ray, K.; Rodgers, D.W.
    Swapping the substrate specificities of the neuropeptidases neurolysin and thimet oligopeptidase (2007), J. Biol. Chem., 282, 9722-9732.
    View publication on PubMed
Search for more literature for 3.4.24.15

Cloned(Commentary)

Cloned (Comment) Organism
expression of wild-type and mutant enzymes in Escherichia coli strain BL21(DE3) Homo sapiens

Crystallization (Commentary)

Crystallization (Comment) Organism
purified recombinant mutant E469R/R498T, hanging drop vapour diffusion method, 4°C, 0.001 ml of 10 mg/ml protein is mixed with 0.001 ml of well solution containing 100 mM sodium cacodylate, pH 6.5, 100 mM magnesium acetate, 2 mM 2-mercaptoethanol, and 12-14% w/v polyethylene glycol 6000, cryoprotection by 25% glycerol, X-ray diffraction structure determination and analysis at 1.94 A resolution Homo sapiens

Protein Variants

Protein Variants Comment Organism
E469R/R498T site-directed mutagenesis of the substrate recognition residues leads to a swap of substrate specificity from thimet oligopeptidase to neurolysin, EC 3.4.24.16, the mutant cleaves neurolysin sites, overview Homo sapiens
H495N/E469R/R498T site-directed mutagenesis of the substrate recognition residues leads to a swap of substrate specificity from thimet oligopeptidase to neurolysin, EC 3.4.24.16, the mutant cleaves neurolysin sites, overview Homo sapiens
M490R/E469R/R498T site-directed mutagenesis of the substrate recognition residues leads to a swap of substrate specificity from thimet oligopeptidase to neurolysin, EC 3.4.24.16, the mutant cleaves neurolysin sites, overview Homo sapiens
M490R/H495N/E469R site-directed mutagenesis of the substrate recognition residues leads to a swap of substrate specificity from thimet oligopeptidase to neurolysin, EC 3.4.24.16, the mutant cleaves TOP and neurolysin sites, overview Homo sapiens
M490R/H495N/R498T site-directed mutagenesis of the substrate recognition residues leads to a swap of substrate specificity from thimet oligopeptidase to neurolysin, EC 3.4.24.16, the mutant cleaves TOP and neurolysin sites, overview Homo sapiens

KM Value [mM]

KM Value [mM] KM Value Maximum [mM] Substrate Comment Organism Structure
0.0026
-
 neurotensin recombinant wild-type enzyme, pH 7.5, 37°C Homo sapiens
0.0033
-
 neurotensin recombinant mutant E469R/R498T, pH 7.5, 37°C Homo sapiens

Metals/Ions

Metals/Ions Comment Organism Structure
Zn2+ a zinc-dependent metallopeptidase Homo sapiens

Natural Substrates/ Products (Substrates)

Natural Substrates Organism Comment (Nat. Sub.) Natural Products Comment (Nat. Pro.) Rev. Reac.
additional information Homo sapiens the enzyme cleaves several bioactive peptides at sites similar or different from neurolysin, EC 3.4.24.16 ?
-
 ?
neurotensin + H2O Homo sapiens
-
 ?
-
 ?

Organism

Organism UniProt Comment Textmining
Homo sapiens
-
-
-

Purification (Commentary)

Purification (Comment) Organism
recombinant wild-type and mutant enzymes from Escherichia coli strain Bl21(DE3) Homo sapiens

Substrates and Products (Substrate)

Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
additional information the enzyme cleaves several bioactive peptides at sites similar or different from neurolysin, EC 3.4.24.16 Homo sapiens ?
-
 ?
additional information residues E469, M490, H495, and R498 determine the substrate specificity of thimet oligopeptidase different from closely related neurolysin, EC 3.4.24.16 Homo sapiens ?
-
 ?
neurotensin + H2O
-
 Homo sapiens ?
-
 ?
neurotensin + H2O determination of specific cleavage sites, overview Homo sapiens ?
-
 ?

Synonyms

Synonyms Comment Organism
TOP
-
 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
2.3
-
 neurotensin recombinant wild-type enzyme, pH 7.5, 37°C Homo sapiens
4.2
-
 neurotensin recombinant mutant E469R/R498T, pH 7.5, 37°C Homo sapiens

pH Optimum

pH Optimum Minimum pH Optimum Maximum Comment Organism
7.5
-
 assay at Homo sapiens