Cloned (Comment) | Organism |
---|---|
recombinant expression of wild-type and mutant enzymes in HEK-293T cells | Homo sapiens |
Protein Variants | Comment | Organism |
---|---|---|
F563I | site-directed mutagenesis, active site mutant which displays an increase in preference towards cleaving leucine5-enkephalin relative to insulin B chain, reduced activity with glutaryl-Ala-Ala-Phe-MNA compared to the wild-type enzyme | Homo sapiens |
F563L | site-directed mutagenesis, active site mutant which exhibits different cleavage site preferences with insulin B chain and amyloid beta1-40 as substrates compared to the wild-type enzyme, similar activity with glutaryl-Ala-Ala-Phe-MNA as the wild-type enzyme | Homo sapiens |
F563M | site-directed mutagenesis, active site mutant which exhibits reduced activity with glutaryl-Ala-Ala-Phe-MNA compared to the wild-type enzyme | Homo sapiens |
F563V | site-directed mutagenesis, active site mutant which exhibits reduced activity with glutaryl-Ala-Ala-Phe-MNA compared to the wild-type enzyme | Homo sapiens |
additional information | it is possible to alter the cleavage site specificity of neprilysin opening the way for the development of substrate specific or substrate exclusive forms of the enzyme with enhanced therapeutic potential | Homo sapiens |
S546A | site-directed mutagenesis, active site mutant with highly reduced activity compared to the wild-type enzyme | Homo sapiens |
S546E | site-directed mutagenesis, active site mutant, that is less discriminating than wild-type neprilysin and exhibits different cleavage site preferences with insulin B chain and amyloid beta1-40 as substrates, reduced activity with glutaryl-Ala-Ala-Phe-MNA compared to the wild-type enzyme | Homo sapiens |
S546T | site-directed mutagenesis, active site mutant with highly reduced activity compared to the wild-type enzyme | Homo sapiens |
Inhibitors | Comment | Organism | Structure |
---|---|---|---|
Insulin B chain | inhibits the activity with substrate N-(4-carboxy-1-oxobutyl)-L-alanyl-L-alanyl-N-(4-methoxy-2-naphthalenyl)-L-phenylalaninamide | Homo sapiens |
KM Value [mM] | KM Value Maximum [mM] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|
0.011 | - |
N-(4-carboxy-1-oxobutyl)-L-alanyl-L-alanyl-N-(4-methoxy-2-naphthalenyl)-L-phenylalaninamide | pH 6.5, 37°C, mutant NEPS546A | Homo sapiens | |
0.051 | - |
N-(4-carboxy-1-oxobutyl)-L-alanyl-L-alanyl-N-(4-methoxy-2-naphthalenyl)-L-phenylalaninamide | pH 6.5, 37°C, wild-type enzyme and mutant NEPF563V | Homo sapiens | |
0.073 | - |
N-(4-carboxy-1-oxobutyl)-L-alanyl-L-alanyl-N-(4-methoxy-2-naphthalenyl)-L-phenylalaninamide | pH 6.5, 37°C, mutant NEPS546T | Homo sapiens | |
0.074 | - |
N-(4-carboxy-1-oxobutyl)-L-alanyl-L-alanyl-N-(4-methoxy-2-naphthalenyl)-L-phenylalaninamide | pH 6.5, 37°C, mutant NEPS546E | Homo sapiens | |
0.081 | - |
N-(4-carboxy-1-oxobutyl)-L-alanyl-L-alanyl-N-(4-methoxy-2-naphthalenyl)-L-phenylalaninamide | pH 6.5, 37°C, mutant NEPF563L | Homo sapiens | |
0.083 | - |
N-(4-carboxy-1-oxobutyl)-L-alanyl-L-alanyl-N-(4-methoxy-2-naphthalenyl)-L-phenylalaninamide | pH 6.5, 37°C, mutant NEPF563I | Homo sapiens | |
0.087 | - |
N-(4-carboxy-1-oxobutyl)-L-alanyl-L-alanyl-N-(4-methoxy-2-naphthalenyl)-L-phenylalaninamide | pH 6.5, 37°C, mutant NEPF563M | Homo sapiens |
Localization | Comment | Organism | GeneOntology No. | Textmining |
---|---|---|---|---|
membrane | membrane-bound | Homo sapiens | 16020 | - |
Natural Substrates | Organism | Comment (Nat. Sub.) | Natural Products | Comment (Nat. Pro.) | Rev. | Reac. |
---|---|---|---|---|---|---|
amyloid beta peptide1-40 + H2O | Homo sapiens | multiple cleavage sites | ? | - |
? | |
amyloid beta peptide1-42 + H2O | Homo sapiens | multiple cleavage sites | ? | - |
? | |
insulin B chain + H2O | Homo sapiens | multiple cleavage sites | ? | - |
? | |
leucine5-enkephalin + H2O | Homo sapiens | Tyr-Gly-Gly-Phe-Leu is cleaved at the Gly-Phe bond by the wild-type enzyme | ? | - |
? | |
additional information | Homo sapiens | the enzyme has broader substrate specificity and is a peptidase capable of cleaving a variety of physiological peptides | ? | - |
? |
Organism | UniProt | Comment | Textmining |
---|---|---|---|
Homo sapiens | P08473 | - |
- |
Specific Activity Minimum [µmol/min/mg] | Specific Activity Maximum [µmol/min/mg] | Comment | Organism |
---|---|---|---|
additional information | - |
activities of recombinant wild-type and mutant enzymes with physiological substrates, overview | Homo sapiens |
Substrates | Comment Substrates | Organism | Products | Comment (Products) | Rev. | Reac. |
---|---|---|---|---|---|---|
amyloid beta peptide + H2O | cleavage sites, overview | Homo sapiens | ? | - |
? | |
amyloid beta peptide1-40 + H2O | multiple cleavage sites | Homo sapiens | ? | - |
? | |
amyloid beta peptide1-40 + H2O | multiple cleavage sites, product peaks corresponding to Abeta1-16, Abeta1-17, Abeta10-17, Abeta20-28, Abeta20-29, and Abeta20-30. The C-terminal products result from the cleavages at K28-G29, G29-A30, and A30-I31. These products are derived from the trans-membrane region of the amyloid precursor protein (APP) from which Abeta is formed and are rather hydrophobic | Homo sapiens | ? | - |
? | |
amyloid beta peptide1-42 + H2O | multiple cleavage sites | Homo sapiens | ? | - |
? | |
insulin B chain + H2O | multiple cleavage sites | Homo sapiens | ? | - |
? | |
insulin B chain + H2O | multiple cleavage sites, product analysis and cleavage sites preferences of wild-type and mutant neprilysins, cleavage sites, overview | Homo sapiens | ? | - |
? | |
leucine5-enkephalin + H2O | Tyr-Gly-Gly-Phe-Leu is cleaved at the Gly-Phe bond by the wild-type enzyme | Homo sapiens | ? | - |
? | |
additional information | the enzyme has broader substrate specificity and is a peptidase capable of cleaving a variety of physiological peptides | Homo sapiens | ? | - |
? | |
N-(4-carboxy-1-oxobutyl)-L-alanyl-L-alanyl-N-(4-methoxy-2-naphthalenyl)-L-phenylalaninamide + H2O | - |
Homo sapiens | N-(4-carboxy-1-oxobutyl)-L-alanyl-L-alanyl-L-phenylalanine + 4-methoxy-2-naphthylamine | - |
? |
Synonyms | Comment | Organism |
---|---|---|
NEP | - |
Homo sapiens |
Temperature Optimum [°C] | Temperature Optimum Maximum [°C] | Comment | Organism |
---|---|---|---|
37 | - |
assay at | Homo sapiens |
pH Optimum Minimum | pH Optimum Maximum | Comment | Organism |
---|---|---|---|
6.5 | - |
assay at | Homo sapiens |
Ki Value [mM] | Ki Value maximum [mM] | Inhibitor | Comment | Organism | Structure |
---|---|---|---|---|---|
0.001 | - |
Insulin B chain | pH 6.5, 37°C, mutant NEPF563V | Homo sapiens | |
0.0012 | - |
Insulin B chain | pH 6.5, 37°C, mutant NEPF563M | Homo sapiens | |
0.0015 | - |
Insulin B chain | pH 6.5, 37°C, mutant NEPF563L | Homo sapiens | |
0.0019 | - |
Insulin B chain | pH 6.5, 37°C, wild-type enzyme | Homo sapiens | |
0.0025 | - |
Insulin B chain | pH 6.5, 37°C, mutant NEPF563I | Homo sapiens |
General Information | Comment | Organism |
---|---|---|
evolution | the enzyme is a member of the M13 subgroup of the zinc-dependent endopeptidase family | Homo sapiens |