Application | Comment | Organism |
---|---|---|
medicine | engineered neprilysin can be a therapeutic for Alzheimer's disease due to its ability to degrade the peptide amyloid beta | Homo sapiens |
Cloned (Comment) | Organism |
---|---|
recombinant expression of His10-tagged wild-type and mutant enzymes in CHO suspension cells, recombinant expression of HA-tagged wild-type and mutant enzymes in Saccharomyces cerevisiae strain YMR307w | Homo sapiens |
Crystallization (Comment) | Organism |
---|---|
purified recombinant enzyme mutant G399V/G714K in complex with inhibitor phorphoramidon, hanging drop vapor diffusion method, mixing of 0.001 ml of 10 mg/ml protein in 25 mM Tris-HCl, pH 7.0, 150 mM NaCl and 2 mM MgCl2 with 2 mM phosphoramidon, 0.001 ml of reservoir solution containing 100 mM HEPES, pH 7.0, 22% w/v PEG3350 and 200 mM NaCl, at 20°C, 2 weeks, rod shaped crystals, improvement by streak seeding, X-ray diffraction structure determination and analysis at 2.15 A resolution, modeling | Homo sapiens |
Protein Variants | Comment | Organism |
---|---|---|
G399V | site-directed mutagenesis, the mutant shows increased catalytic efficiency on Ab1-40 with 6fold increased catalytic efficiency compared to the wild-type enzyme. The G399V mutation also significantly reduces the catalytic efficiency on angiotensin, bradykinin and neurotensin compared to the wild-type enzyme | Homo sapiens |
G399V/G714K | site-directed mutagenesis, the enzyme variant displays an approximately 20fold improved activity on amyloid beta 1-40 and up to a 3200fold reduction in activity on other peptides, and the mutant enzyme produces a markedly altered series of amyloid beta cleavage products compared to the wild-type enzyme | Homo sapiens |
G714K | site-directed mutagenesis, the mutant shows increased catalytic efficiency on Ab1-40 with 6fold increased catalytic efficiency compared to the wild-type enzyme | Homo sapiens |
additional information | generation of a soluble version of the ectodomain of neprilysin with improved activity and specificity towards amyloid beta peptide as a potential therapeutic for Alzheimer's disease | Homo sapiens |
Inhibitors | Comment | Organism | Structure |
---|---|---|---|
phosphoramidon | - |
Homo sapiens | |
thiorphan | - |
Homo sapiens |
KM Value [mM] | KM Value Maximum [mM] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|
additional information | - |
additional information | Michaelis-Menten profiles and steady-state kinetic parameters for cleavage of amyloid beta peptide1-40 and off-target peptide by wild-type and mutant enzymes, overview | Homo sapiens | |
0.0062 | - |
Arg-vasopressin | wild-type enzyme, pH and temperature not specified in the publication | Homo sapiens | |
0.0074 | - |
somatostatin 14 | wild-type enzyme, pH and temperature not specified in the publication | Homo sapiens | |
0.0086 | - |
angiotensin | wild-type enzyme, pH and temperature not specified in the publication | Homo sapiens | |
0.0093 | - |
Neurokinin A | wild-type enzyme, pH and temperature not specified in the publication | Homo sapiens | |
0.013 | - |
gamma-endorphin | wild-type enzyme, pH and temperature not specified in the publication | Homo sapiens | |
0.014 | - |
amyloid beta peptide1-40 | mutant G399V/G714K, pH and temperature not specified in the publication | Homo sapiens | |
0.016 | - |
Substance P | wild-type enzyme, pH and temperature not specified in the publication | Homo sapiens | |
0.016 | - |
neurotensin | wild-type enzyme, pH and temperature not specified in the publication | Homo sapiens | |
0.02 | - |
nociceptin | wild-type enzyme, pH and temperature not specified in the publication | Homo sapiens | |
0.022 | - |
neurokinin B | wild-type enzyme, pH and temperature not specified in the publication | Homo sapiens | |
0.024 | - |
GLP-1 | wild-type enzyme, pH and temperature not specified in the publication | Homo sapiens | |
0.024 | - |
alpha-endorphin | wild-type enzyme, pH and temperature not specified in the publication | Homo sapiens | |
0.025 | - |
glucagon | wild-type enzyme, pH and temperature not specified in the publication | Homo sapiens | |
0.031 | - |
somatostatin 28 | wild-type enzyme, pH and temperature not specified in the publication | Homo sapiens | |
0.032 | - |
bradykinin | wild-type enzyme, pH and temperature not specified in the publication | Homo sapiens | |
0.042 | - |
amyloid beta peptide1-40 | mutant G714K, pH and temperature not specified in the publication | Homo sapiens | |
0.053 | - |
endothelin-1 | wild-type enzyme, pH and temperature not specified in the publication | Homo sapiens | |
0.055 | - |
amyloid beta peptide1-40 | mutant G399V, pH and temperature not specified in the publication | Homo sapiens | |
0.104 | - |
amyloid beta peptide1-40 | wild-type enzyme, pH and temperature not specified in the publication | Homo sapiens | |
0.15 | - |
gastri-releasing peptide | wild-type enzyme, pH and temperature not specified in the publication | Homo sapiens |
Localization | Comment | Organism | GeneOntology No. | Textmining |
---|---|---|---|---|
membrane | a transmembrane zinc metallopeptidase | Homo sapiens | 16020 | - |
Metals/Ions | Comment | Organism | Structure |
---|---|---|---|
Zn2+ | a transmembrane zinc metallopeptidase | Homo sapiens |
Molecular Weight [Da] | Molecular Weight Maximum [Da] | Comment | Organism |
---|---|---|---|
82000 | 82410 | recombinant enzyme, gel filtration | Homo sapiens |
Natural Substrates | Organism | Comment (Nat. Sub.) | Natural Products | Comment (Nat. Pro.) | Rev. | Reac. |
---|---|---|---|---|---|---|
amyloid beta peptide + H2O | Homo sapiens | degradation | ? | - |
? | |
amyloid beta peptide1-40 + H2O | Homo sapiens | degradation | ? | - |
? | |
amyloid beta peptide1-42 + H2O | Homo sapiens | degradation | ? | - |
? | |
additional information | Homo sapiens | the enzyme degrades a wide range of peptide substrates, physiologically relevant peptides are such as enkephalins, tachykinins, and natriuretic peptides | ? | - |
? |
Organism | UniProt | Comment | Textmining |
---|---|---|---|
Homo sapiens | P08473 | - |
- |
Purification (Comment) | Organism |
---|---|
recombinant His10-tagged wild-type and mutant enzymes from CHO suspension cells by nickel affinity chromatography, recombinant HA-tagged wild-type and mutant enzymes from Saccharomyces cerevisiae strain YMR307w by immunoaffinity chromatography | Homo sapiens |
Substrates | Comment Substrates | Organism | Products | Comment (Products) | Rev. | Reac. |
---|---|---|---|---|---|---|
alpha-endorphin + H2O | - |
Homo sapiens | ? | - |
? | |
amyloid beta peptide + H2O | - |
Homo sapiens | ? | - |
? | |
amyloid beta peptide + H2O | degradation | Homo sapiens | ? | - |
? | |
amyloid beta peptide1-40 + H2O | degradation | Homo sapiens | ? | - |
? | |
amyloid beta peptide1-40 + H2O | the wild-type enzyme cleaves Ab1-40 predominantly at Lys16-Leu17, Leu17-Val18 and Phe19-Phe20. The mutant G399V/G714K cleaves preferentially at Phe20-Ala21 | Homo sapiens | amyloid beta peptide Asp1-Lys16 + amyloid beta peptide Asp1-Leu17 + amyloid beta peptide Asp1-Phe19 | main cleavage fragments after 60 min with the wild-type enzyme. After 360 min these fragments are degraded further, accompanied by the appearance of Val12-Leu17 and Tyr10-Leu17 fragments | ? | |
amyloid beta peptide1-42 + H2O | - |
Homo sapiens | ? | - |
? | |
amyloid beta peptide1-42 + H2O | degradation | Homo sapiens | ? | - |
? | |
angiotensin + H2O | - |
Homo sapiens | ? | - |
? | |
Arg-vasopressin + H2O | - |
Homo sapiens | ? | - |
? | |
bradykinin + H2O | - |
Homo sapiens | ? | - |
? | |
endothelin-1 + H2O | - |
Homo sapiens | ? | - |
? | |
gamma-endorphin + H2O | - |
Homo sapiens | ? | - |
? | |
gastri-releasing peptide + H2O | - |
Homo sapiens | ? | - |
? | |
GLP-1 + H2O | - |
Homo sapiens | ? | - |
? | |
Glucagon + H2O | - |
Homo sapiens | ? | - |
? | |
additional information | the enzyme degrades a wide range of peptide substrates, physiologically relevant peptides are such as enkephalins, tachykinins, and natriuretic peptides | Homo sapiens | ? | - |
? | |
Neurokinin A + H2O | - |
Homo sapiens | ? | - |
? | |
neurokinin B + H2O | - |
Homo sapiens | ? | - |
? | |
neurotensin + H2O | - |
Homo sapiens | ? | - |
? | |
nociceptin + H2O | - |
Homo sapiens | ? | - |
? | |
somatostatin 14 + H2O | - |
Homo sapiens | ? | - |
? | |
somatostatin 28 + H2O | - |
Homo sapiens | ? | - |
? | |
Substance P + H2O | - |
Homo sapiens | ? | - |
? |
Synonyms | Comment | Organism |
---|---|---|
NEP | - |
Homo sapiens |
neutral endopeptidase | - |
Homo sapiens |
Turnover Number Minimum [1/s] | Turnover Number Maximum [1/s] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|
0.68 | - |
endothelin-1 | wild-type enzyme, pH and temperature not specified in the publication | Homo sapiens | |
0.83 | - |
amyloid beta peptide1-40 | wild-type enzyme, pH and temperature not specified in the publication | Homo sapiens | |
1.9 | - |
amyloid beta peptide1-40 | mutant G714K, pH and temperature not specified in the publication | Homo sapiens | |
2.1 | - |
amyloid beta peptide1-40 | mutant G399V/G714K, pH and temperature not specified in the publication | Homo sapiens | |
2.7 | - |
amyloid beta peptide1-40 | mutant G399V, pH and temperature not specified in the publication | Homo sapiens | |
2.8 | - |
Arg-vasopressin | wild-type enzyme, pH and temperature not specified in the publication | Homo sapiens | |
3.3 | - |
GLP-1 | wild-type enzyme, pH and temperature not specified in the publication | Homo sapiens | |
4.5 | - |
glucagon | wild-type enzyme, pH and temperature not specified in the publication | Homo sapiens | |
4.8 | - |
gamma-endorphin | wild-type enzyme, pH and temperature not specified in the publication | Homo sapiens | |
8.2 | - |
angiotensin | wild-type enzyme, pH and temperature not specified in the publication | Homo sapiens | |
8.9 | - |
alpha-endorphin | wild-type enzyme, pH and temperature not specified in the publication | Homo sapiens | |
9.1 | - |
somatostatin 28 | wild-type enzyme, pH and temperature not specified in the publication | Homo sapiens | |
11 | - |
nociceptin | wild-type enzyme, pH and temperature not specified in the publication | Homo sapiens | |
13 | - |
neurotensin | wild-type enzyme, pH and temperature not specified in the publication | Homo sapiens | |
16 | - |
bradykinin | wild-type enzyme, pH and temperature not specified in the publication | Homo sapiens | |
17 | - |
Neurokinin A | wild-type enzyme, pH and temperature not specified in the publication | Homo sapiens | |
18 | - |
Substance P | wild-type enzyme, pH and temperature not specified in the publication | Homo sapiens | |
20 | - |
neurokinin B | wild-type enzyme, pH and temperature not specified in the publication | Homo sapiens | |
26 | - |
gastri-releasing peptide | wild-type enzyme, pH and temperature not specified in the publication | Homo sapiens | |
32 | - |
somatostatin 14 | wild-type enzyme, pH and temperature not specified in the publication | Homo sapiens |
IC50 Value | IC50 Value Maximum | Comment | Organism | Inhibitor | Structure |
---|---|---|---|---|---|
0.00001 | - |
wild-type enzyme, pH and temperature not specified in the publication | Homo sapiens | phosphoramidon | |
0.00001 | - |
below, wild-type enzyme, pH and temperature not specified in the publication | Homo sapiens | thiorphan | |
0.000075 | - |
enzyme mutant G714K, pH and temperature not specified in the publication | Homo sapiens | phosphoramidon | |
0.0044 | - |
enzyme mutant G399V, pH and temperature not specified in the publication | Homo sapiens | phosphoramidon | |
0.021 | - |
enzyme mutant G399V/G714K, pH and temperature not specified in the publication | Homo sapiens | phosphoramidon | |
0.077 | - |
enzyme mutant G399V/G714K, pH and temperature not specified in the publication | Homo sapiens | thiorphan |
General Information | Comment | Organism |
---|---|---|
evolution | the enzyme harbors an ectodomain, which contains the catalytic site and belongs to the M13 family of peptidases/proteases | Homo sapiens |
additional information | the enzyme is composed of an ectodomain, which contains the catalytic site, a transmembrane domain and a short intracellular domain. The structure of the ectodomain is composed of two largely alpha-helical domains that are arranged to form a central spherical water-filled core that contains the active site of the enzyme | Homo sapiens |
physiological function | the enzyme degrades a number of physiological peptides that are involved in processes such as blood pressure regulation and nociception | Homo sapiens |
kcat/KM Value [1/mMs-1] | kcat/KM Value Maximum [1/mMs-1] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|
8 | - |
amyloid beta peptide1-40 | wild-type enzyme, pH and temperature not specified in the publication | Homo sapiens | |
13 | - |
endothelin-1 | wild-type enzyme, pH and temperature not specified in the publication | Homo sapiens | |
45 | - |
amyloid beta peptide1-40 | mutant G714K, pH and temperature not specified in the publication | Homo sapiens | |
49 | - |
amyloid beta peptide1-40 | mutant G399V, pH and temperature not specified in the publication | Homo sapiens | |
140 | - |
GLP-1 | wild-type enzyme, pH and temperature not specified in the publication | Homo sapiens | |
150 | - |
amyloid beta peptide1-40 | mutant G399V/G714K, pH and temperature not specified in the publication | Homo sapiens | |
160 | - |
somatostatin 28 | wild-type enzyme, pH and temperature not specified in the publication | Homo sapiens | |
170 | - |
gastri-releasing peptide | wild-type enzyme, pH and temperature not specified in the publication | Homo sapiens | |
180 | - |
glucagon | wild-type enzyme, pH and temperature not specified in the publication | Homo sapiens | |
380 | - |
gamma-endorphin | wild-type enzyme, pH and temperature not specified in the publication | Homo sapiens | |
390 | - |
alpha-endorphin | wild-type enzyme, pH and temperature not specified in the publication | Homo sapiens | |
450 | - |
Arg-vasopressin | wild-type enzyme, pH and temperature not specified in the publication | Homo sapiens | |
480 | - |
bradykinin | wild-type enzyme, pH and temperature not specified in the publication | Homo sapiens | |
570 | - |
nociceptin | wild-type enzyme, pH and temperature not specified in the publication | Homo sapiens | |
820 | - |
neurotensin | wild-type enzyme, pH and temperature not specified in the publication | Homo sapiens | |
920 | - |
neurokinin B | wild-type enzyme, pH and temperature not specified in the publication | Homo sapiens | |
970 | - |
angiotensin | wild-type enzyme, pH and temperature not specified in the publication | Homo sapiens | |
1100 | - |
Substance P | wild-type enzyme, pH and temperature not specified in the publication | Homo sapiens | |
1700 | - |
Neurokinin A | wild-type enzyme, pH and temperature not specified in the publication | Homo sapiens | |
4400 | - |
somatostatin 14 | wild-type enzyme, pH and temperature not specified in the publication | Homo sapiens |