Information on EC 3.4.24.B14 - neprilysin-2

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The enzyme appears in viruses and cellular organisms

EC NUMBER
COMMENTARY hide
3.4.24.B14
preliminary BRENDA-supplied EC number
RECOMMENDED NAME
GeneOntology No.
neprilysin-2
-
REACTION
REACTION DIAGRAM
COMMENTARY hide
ORGANISM
UNIPROT
LITERATURE
proteolytic degradation of proteins
show the reaction diagram
-
-
-
-
REACTION TYPE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
hydrolysis of peptide bond
-
-
CAS REGISTRY NUMBER
COMMENTARY hide
252986-92-8
-
82707-54-8
not distinguished from EC 3.4.24.11
ORGANISM
COMMENTARY hide
LITERATURE
UNIPROT
SEQUENCE DB
SOURCE
strain OregonR
-
-
Manually annotated by BRENDA team
GENERAL INFORMATION
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
evolution
malfunction
metabolism
physiological function
additional information
SUBSTRATE
PRODUCT                       
REACTION DIAGRAM
ORGANISM
UNIPROT
COMMENTARY
(Substrate) hide
LITERATURE
(Substrate)
COMMENTARY
(Product) hide
LITERATURE
(Product)
Reversibility
r=reversible
ir=irreversible
?=not specified
3-methoxysuccinyl-Gly-Leu-Phe-Met-7-amido-4-methylcoumarin + H2O
3-methoxysuccinyl-Gly-Leu + Phe-Met-7-amido-4-methylcoumarin
show the reaction diagram
-
-
-
-
?
amyloid beta peptide + H2O
?
show the reaction diagram
-
-
-
-
?
amyloid beta peptide1-42 + H2O
?
show the reaction diagram
-
-
-
-
?
Angiotensin + H2O
?
show the reaction diagram
-
cleavage sites: DRVY-/-IHP-/-FHL
-
-
?
Angiotensin I + H2O
?
show the reaction diagram
Arg-Pro-Lys-Pro-Gln-Gln-Phe-Phe-Gly-Leu-Met-NH2 + H2O
Arg-Pro-Lys-Pro-Gln-Gln + Phe-Phe-Gly + Leu-Met-NH2
show the reaction diagram
-
i.e. substance P(1-11), Arg-Pro-Lys-Pro-Gln-Gln-/-Phe-Phe-Gly-/-Leu-Met-NH2. Hydrolysis of substance P and derivatives occurs predominantly at the Gly9-/-Leu10 bond in the case of neprilysin and neprilysin 2. In addition neprilysin 2 cleaves at the Gln6-Phe7 bond
-
-
?
Arg-Pro-Pro-Gly-Phe-Ser-Pro-Phe-Arg + H2O
Arg-Pro-Pro-Gly-Phe-Ser-Pro + Phe-Arg
show the reaction diagram
-
i.e. bradykinin
-
-
?
Asp-Arg-Val-Tyr-Ile-His-Pro-Phe-His-Leu + H2O
Asp-Arg-Val-Tyr-Ile-His-Pro + Phe-His-Leu
show the reaction diagram
-
i.e. angiotensin
-
-
?
Asp-Tyr(sulfated)-Met-Gly-Trp-Met-Asp-Phe-NH2 + H2O
Asp-Tyr(sulfated)-Met-Gly + Trp-Met-Asp-Phe-NH2
show the reaction diagram
-
i.e. cholecystokinin-8(sulfated). In cholecystokinins and derivatives, clear differences between neprilysin (EC 3.4.24.11) and neprilysin 2 can be observed: NEP2 does not attack the Asp7-/-Phe8-NH2 bond, a preferential cleavage site for neprilysin (EC 3.4.24.11, NEP) but cleaves the Gly4-/-Trp5 amide bond
-
-
?
Asp-Tyr-Met-Gly-Trp-Met + H2O
Asp-Tyr-Met-Gly + Trp-Met
show the reaction diagram
-
i.e. cholecystokinin-(1-6)
-
-
?
Asp-Tyr-Met-Gly-Trp-Met-Asp-Phe-NH2 + H2O
Asp-Tyr-Met-Gly + Trp-Met-Asp-Phe-NH2
show the reaction diagram
-
i.e. cholecystokinin-8(non-sulfated)
-
-
?
atrial natriuretic peptide + H2O
?
show the reaction diagram
-
-
?
Bradykinin + H2O
?
show the reaction diagram
cholecystokinin(1-6) + H2O
?
show the reaction diagram
-
cleavage site: DYMG-/-WM
-
-
?
cholecystokinin-8 + H2O
?
show the reaction diagram
-
cleavage site: DYMG-/-WMDF-NH2
-
-
?
Drosophila tachykinin-1 + H2O
APTSS + FIGMR-amide
show the reaction diagram
Drosophila tachykinin-2 + H2O
APLAFYG + Leu-Arg-amide
show the reaction diagram
Drosophila tachykinin-3 + H2O
APTG + FTGMR-amide
show the reaction diagram
Drosophila tachykinin-4 + H2O
APVNS + FVGMR-amide
show the reaction diagram
Drosophila tachykinin-5 + H2O
APNG + FLGMR-amide
show the reaction diagram
Drosophila tachykinin-6 + H2O
QQR + Phe + ADFNSKFVAVR-amide
show the reaction diagram
-
17% activity compared to Locusta tachykinin-1
-
-
?
endothelin-1 + H2O
?
show the reaction diagram
-
-
?
Gly-Gly-Phe-Met + H2O
Gly-Gly + Phe-Met
show the reaction diagram
-
i.e. Des-[Tyr1, Met5]enkephalin
-
-
?
Gly-Trp-Met-Asp-Phe-NH2 + H2O
Gly + Trp-Met-Asp-Phe-NH2
show the reaction diagram
-
i.e. cholecystokinin-(4-8)
-
-
?
gonadotropin-releasing hormone + H2O
?
show the reaction diagram
GPSGFYGVR-NH2 + H2O
GPSGFYG + VR-NH2
show the reaction diagram
-
-
-
?
Leu5-enkephalin + H2O
?
show the reaction diagram
-
cleavage site: YGG-/-FL
-
-
?
Leu5-enkephalin-NH2 + H2O
?
show the reaction diagram
-
cleavage site: YGG-/-FL-NH2
-
-
?
Locusta tachykinin-1 + H2O
GPSGFYG + Val-amide
show the reaction diagram
-
100% activity
-
-
?
locustatachykinin 1 + H2O
?
show the reaction diagram
-
-
-
-
?
NEP fluorogenic substrate + H2O
?
show the reaction diagram
hydrolyzed to 39%
-
-
?
Phe-Gly-Leu-Met-NH2 + H2O
Phe-Gly + Leu-Met-NH2
show the reaction diagram
-
i.e. substance P(8-11)
-
-
?
Phe-Phe-Gly-Leu-Met-NH2 + H2O
Phe-Phe-Gly + Leu-Met-NH2
show the reaction diagram
-
i.e. substance P(7-11)
-
-
?
protein + H2O
peptides
show the reaction diagram
Substance P + H2O
?
show the reaction diagram
succinyl-Ala-Ala-Phe-4-methylcoumarin 7-amide + H2O
succinyl-Ala-Ala + Phe-4-methylcoumarin 7-amide
show the reaction diagram
-
-
-
-
?
succinyl-Ala-Ala-Phe-7-amido-4-methylcoumarin + H2O
succinyl-Ala-Ala + Phe-7-amido-4-methylcoumarin
show the reaction diagram
succinyl-Ala-Ala-Val-7-amido-4-methylcoumarin + H2O
succinyl-Ala-Ala + Val-7-amido-4-methylcoumarin
show the reaction diagram
-
-
-
-
?
Tyr-Gly-Ala-Phe-Met + H2O
Tyr-Gly-Ala + Phe-Met
show the reaction diagram
-
i.e. [L-Ala3,Met5]enkephalin
-
-
?
Tyr-Gly-Gly-Phe-Leu + H2O
Tyr-Gly-Gly + Phe-Leu
show the reaction diagram
-
i.e. [Leu5]enkephalin
-
-
?
Tyr-Gly-Gly-Phe-Leu-NH2 + H2O
Tyr-Gly-Gly + Phe-Leu-NH2
show the reaction diagram
-
i.e. [Leu5]enkephalin-NH2
-
-
?
Tyr-Gly-Gly-Phe-Met + H2O
Tyr-Gly-Gly + Phe-Met
show the reaction diagram
-
i.e. [Met5]enkephalin. Regarding enkephalins and related opioid peptides, neprilysin (EC 3.4.24.11, NEP) and neprilysin 2 (NEP2) cleave the same Gly-/-Phe bond in all susceptible peptides, and hydrolysis by NEP2 and NEP is strongly reduced when the C-terminal carboxylate is amidated. Differences are: Ala instead of Gly in P1 significantly enhances hydrolysis by NEP2 but not NEP, whereas Leu instead of Met in P2' has an opposite influence (whereas affinity is enhanced) on NEP2. In contrast, neither affinity nor hydrolysis was significantly affected by the latter change in the case of NEP (EC 3.4.24.11)
-
-
?
Tyr-Gly-Gly-Phe-Met-NH2 + H2O
Tyr-Gly-Gly + Phe-Met-NH2
show the reaction diagram
-
i.e. [Met5]enkephalin-NH2
-
-
?
additional information
?
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NATURAL SUBSTRATES
NATURAL PRODUCTS
REACTION DIAGRAM
ORGANISM
UNIPROT
COMMENTARY
(Substrate) hide
LITERATURE
(Substrate)
COMMENTARY
(Product) hide
LITERATURE
(Product)
REVERSIBILITY
r=reversible
ir=irreversible
?=not specified
amyloid beta peptide + H2O
?
show the reaction diagram
-
-
-
-
?
amyloid beta peptide1-42 + H2O
?
show the reaction diagram
-
-
-
-
?
gonadotropin-releasing hormone + H2O
?
show the reaction diagram
Q495T6
-
-
-
?
protein + H2O
peptides
show the reaction diagram
additional information
?
-
METALS and IONS
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
INHIBITORS
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
1,10-phenanthroline
inhibition of the soluble isozyme SEP
angiotensin
-
-
Arg-Pro-Lys-Pro-Gln-Gln-Phe-Phe-Gly-Leu-Met-NH2
-
-
Arg-Pro-Pro-Gly-Phe-Ser-Pro-Phe-Arg
-
-
Asp-Arg-Val-Tyr-Ile-His-Pro-Phe-His-Leu
-
-
Asp-Tyr(sulfated)-Met-Gly-Trp-Met-Asp-Phe-NH2
-
-
Asp-Tyr-Met-Gly-Trp-Met
-
-
Asp-Tyr-Met-Gly-Trp-Met-Asp-Phe-NH2
-
-
bradykinin
-
-
calcium ionophore A23187
-
0.001 mM
cholecystokinin(1-6)
-
-
cholecystokinin-8
-
-
EDTA
-
1 mM, complete inhibition
fasidotrilat
-
-
forskolin
-
0.01 mM
Gly-Gly-Phe-Met
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-
gonadotropin-releasing hormone
-
-
Leu-Met-NH2
-
-
Leu5-enkephalin
-
-
Leu5-enkephalin-NH2
-
-
omapatrilat
-
-
Phe-Phe-Gly-Leu-Met-NH2
-
-
phosphoramidon
Substance P
-
-
succinyl-Ala-Ala-Phe-4-methylcoumarin 7-amide
-
37°C, pH 7.2, wild-type enzyme
thapsigargin
-
0.0001 mM
thiorphan
Trp-Met-Asp-Phe-NH2
-
-
tunicamycin
-
inhibition of N-glycosylation by tunicamycin reduces the enzymatic activity of extracellular NEP2 to about 50% and the molecular size of intracellular NEP2
Tyr-Gly-Ala-Phe-Met
-
-
Tyr-Gly-Gly-Phe-Leu
-
-
Tyr-Gly-Gly-Phe-Leu-NH2
-
-
Tyr-Gly-Gly-Phe-Met
-
-
Tyr-Gly-Gly-Phe-Met-NH2
-
-
Val-Phe-NH2
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-
additional information
-
KM VALUE [mM]
SUBSTRATE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
0.023 - 0.035
3-methoxysuccinyl-Gly-Leu-Phe-Met-7-amido-4-methylcoumarin
0.04 - 1
succinyl-Ala-Ala-Phe-4-methylcoumarin 7-amide
0.027 - 0.05
succinyl-Ala-Ala-Phe-7-amido-4-methylcoumarin
0.04 - 0.1
succinyl-Ala-Ala-Val-7-amido-4-methylcoumarin
TURNOVER NUMBER [1/s]
SUBSTRATE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
11.5
3-methoxysuccinyl-Gly-Leu-Phe-Met-7-amido-4-methylcoumarin
Rattus norvegicus
-
37°C
6
Arg-Pro-Lys-Pro-Gln-Gln-Phe-Phe-Gly-Leu-Met-NH2
Rattus norvegicus
-
37°C
2.5
Arg-Pro-Pro-Gly-Phe-Ser-Pro-Phe-Arg
Rattus norvegicus
-
37°C
3
Asp-Arg-Val-Tyr-Ile-His-Pro-Phe-His-Leu
Rattus norvegicus
-
37°C
6.17
Asp-Tyr(sulfated)-Met-Gly-Trp-Met-Asp-Phe-NH2
Rattus norvegicus
-
37°C
34.5
Asp-Tyr-Met-Gly-Trp-Met
Rattus norvegicus
-
37°C
13.17
Asp-Tyr-Met-Gly-Trp-Met-Asp-Phe-NH2
Rattus norvegicus
-
37°C
8.5
Gly-Gly-Phe-Met
Rattus norvegicus
-
37°C
4
Gly-Trp-Met-Asp-Phe-NH2
Rattus norvegicus
-
37°C
8.5
gonadotropin-releasing hormone
Rattus norvegicus
-
37°C
22.83
Phe-Gly-Leu-Met-NH2
Rattus norvegicus
-
37°C
50
Phe-Phe-Gly-Leu-Met-NH2
Rattus norvegicus
-
37°C
0.52 - 30.7
succinyl-Ala-Ala-Phe-4-methylcoumarin 7-amide
8.5
succinyl-Ala-Ala-Phe-7-amido-4-methylcoumarin
Rattus norvegicus
-
37°C
13.8
succinyl-Ala-Ala-Val-7-amido-4-methylcoumarin
Rattus norvegicus
-
37°C
41.7
Tyr-Gly-Ala-Phe-Met
Rattus norvegicus
-
37°C
2.7
Tyr-Gly-Gly-Phe-Leu
Rattus norvegicus
-
37°C
0.33
Tyr-Gly-Gly-Phe-Leu-NH2
Rattus norvegicus
-
37°C
9
Tyr-Gly-Gly-Phe-Met
Rattus norvegicus
-
37°C
0.33
Tyr-Gly-Gly-Phe-Met-NH2
Rattus norvegicus
-
37°C
Ki VALUE [mM]
INHIBITOR
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
0.008
angiotensin
-
37°C, pH 7.2, wild-type enzyme
0.004
Arg-Pro-Lys-Pro-Gln-Gln-Phe-Phe-Gly-Leu-Met-NH2
-
-
0.002
Arg-Pro-Pro-Gly-Phe-Ser-Pro-Phe-Arg
-
37°C
0.006
Asp-Arg-Val-Tyr-Ile-His-Pro-Phe-His-Leu
-
37°C
0.021
Asp-Tyr(sulfated)-Met-Gly-Trp-Met-Asp-Phe-NH2
-
37°C
0.018
Asp-Tyr-Met-Gly-Trp-Met
-
37°C
0.05
Asp-Tyr-Met-Gly-Trp-Met-Asp-Phe-NH2
-
37°C
0.003
bradykinin
-
37°C, pH 7.2, wild-type enzyme
0.05
cholecystokinin(1-6)
-
37°C, pH 7.2, wild-type enzyme
0.01
cholecystokinin-8
-
37°C, pH 7.2, wild-type enzyme
0.000016 - 0.000019
fasidotrilat
0.014
Gly-Gly-Phe-Met
-
37°C
0.018
gonadotropin-releasing hormone
-
37°C
0.02
Leu-Met
-
37°C, pH 7.2, wild-type enzyme
0.175
Leu-Met-NH2
-
37°C, pH 7.2, wild-type enzyme
0.002
Leu5-enkephalin
-
37°C, pH 7.2, wild-type enzyme
0.02
Leu5-enkephalin-NH2
-
37°C, pH 7.2, wild-type enzyme
0.000008 - 0.000017
omapatrilat
0.09
Phe-Phe-Gly-Leu-Met-NH2
-
-
0.0000008 - 0.000002
phosphoramidon
0.011
Substance P
-
37°C, pH 7.2, wild-type enzyme
0.00012 - 0.00025
thiorphan
0.039
Trp-Met-Asp-Phe-NH2
-
37°C
0.03
Tyr-Gly-Ala-Phe-Met
-
37°C
0.0035
Tyr-Gly-Gly-Phe-Leu
-
37°C
0.021
Tyr-Gly-Gly-Phe-Leu-NH2
-
37°C
0.01
Tyr-Gly-Gly-Phe-Met
-
37°C
0.025
Tyr-Gly-Gly-Phe-Met-NH2
-
37°C
0.021
Val-Phe
-
37°C, pH 7.2, wild-type enzyme
0.13
Val-Phe-NH2
-
37°C, pH 7.2, wild-type enzyme
IC50 VALUE [mM]
INHIBITOR
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
0.000169 - 0.003
phosphoramidon
0.0005 - 0.08
thiorphan
pH OPTIMUM
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
7.4
sharp pH-dependence
TEMPERATURE OPTIMUM
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
37
-
assay at
SOURCE TISSUE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
SOURCE
isozymes SEP and SEPDELTA
Manually annotated by BRENDA team
-
peripheral
Manually annotated by BRENDA team
-
high enzyme level
Manually annotated by BRENDA team
isozymes SEP and SEPDELTA
Manually annotated by BRENDA team
-
high enzyme level
Manually annotated by BRENDA team
isozymes SEP and SEPDELTA
Manually annotated by BRENDA team
isozymes SEP and SEPDELTA
Manually annotated by BRENDA team
isozymes SEP and SEPDELTA
Manually annotated by BRENDA team
isozymes SEP and SEPDELTA
Manually annotated by BRENDA team
isozyme SEP
Manually annotated by BRENDA team
additional information
in most cases isozyme SEPDELTA shows higher expression rates than isozyme SEP
Manually annotated by BRENDA team
LOCALIZATION
ORGANISM
UNIPROT
COMMENTARY hide
GeneOntology No.
LITERATURE
SOURCE
-
human neprilysin-2-beta is either localized to the extracellular surface or secreted
Manually annotated by BRENDA team
additional information
MOLECULAR WEIGHT
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
83000
-
deglycosylated enzyme, SDS-PAGE
100000
-
soluble enzyme, SDS-PAGE
110000
x * 110000, membrane-bound isozymes SEP and SEPDELTA, SDS-PAGE, x * 126000, soluble isozyme SEP, SDS-PAGE
126000
x * 110000, membrane-bound isozymes SEP and SEPDELTA, SDS-PAGE, x * 126000, soluble isozyme SEP, SDS-PAGE
SUBUNITS
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
?
x * 110000, membrane-bound isozymes SEP and SEPDELTA, SDS-PAGE, x * 126000, soluble isozyme SEP, SDS-PAGE
POSTTRANSLATIONAL MODIFICATION
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
glycoprotein
pH STABILITY
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
6.5 - 8
-
activity is drastically reduced below pH 6.5 and above pH 8.0
682251
Purification/COMMENTARY
ORGANISM
UNIPROT
LITERATURE
Phenyl-Sepharose column chromatography and HiTrap Q HP column chromatography
-
Cloned/COMMENTARY
ORGANISM
UNIPROT
LITERATURE
expressed in HEK293 cells
-
expressed in S2 cells
-
expression in Drosophila melanogaster S2 cells
-
expression of both isoforms, NEP2 and NEP2DELTA
expression of two alternatively spliced isoforms of NEP2 in Chinese hamster ovary and AtT20 cells
-
expression of wild-type and mutated forms of rNEP2 in AtT20 cells
-
isozymes SEP and SEPDELTA, DNA determination and analysis, transient expression of both isoforms in CHO cells
neprilysin-2 expression in human induced pluripotent stem (iPS) cell-derived macrophage-like cells which leads to secretion of the enzyme, to reduced level of soluble amyloid beta oligomer in the culture medium, and to alleviated neurotoxicity of amyloid beta. Intracerebrally administration of the iPS-ML/NEP2 cells to 5XFAD mice leading to significant reduction in the level of amyloid beta in the brain interstitial fluid. 5XFAD mice harbor three mutations in the amyloid precursor protein (APP) gene and two in the presenilin-1 (PS1) gene and are a mouse model of Alzheimer's disease
-
EXPRESSION
ORGANISM
UNIPROT
LITERATURE
NEP2 expression is measured in non-impaired, mild cognitive impaired, and clinical Alzheimer's disease human brain tissue from two Alzheimer's disease -susceptible and two resistant regions. NEP2 and NEP mRNA expression is altered in mild cognitive impaired subjects relative to non-impaired subjects in Alzheimer's disease -susceptible regions. NEP2 enzymatic activity is lowered in association with mild cognitive impaired and Alzheimer's disease and is positively associated with cognitive function, independent of diagnostic category
-
the altered enzyme expression has a sexually dimorphic change, with mild-cognitive impaired males having significantly elevated NEP2 mRNA levels compared to non-impaired males in the mid-temporal and mid-frontal gyri
-
ENGINEERING
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
P751A
-
no hydrolysis of succinyl-Ala-Ala-Phe-4-methylcoumarin 7-amide
P764Y
-
kcat/Km is 2.6fold lower than wild-type enzyme
W774A
-
kcat/Km is 1.1fold lower than wild-type enzyme
W774F
-
kcat/Km is nearly identical to wild-type value
APPLICATION
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
medicine
-
altered NEP2 expression in mild cognitive impaired patients may potentially serve as preclinical markers for Alzheimer's disease and reduced NEP2 activity may be associated with the development of Alzheimer's disease