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Literature summary for 3.4.24.11 extracted from

  • Sexton, T.; Hitchcook, L.J.; Rodgers, D.W.; Bradley, L.H.; Hersh, L.B.
    Active site mutations change the cleavage specificity of neprilysin (2012), PLoS ONE, 7, e32343.
    View publication on PubMedView publication on EuropePMC
Search for more literature for 3.4.24.11

Cloned(Commentary)

Cloned (Comment) Organism
recombinant expression of wild-type and mutant enzymes in HEK-293T cells Homo sapiens

Protein Variants

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

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 [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

Localization Comment Organism GeneOntology No. Textmining
membrane membrane-bound Homo sapiens 16020
-

Natural Substrates/ Products (Substrates)

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

Organism UniProt Comment Textmining
Homo sapiens P08473
-
-

Specific Activity [micromol/min/mg]

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 and Products (Substrate)

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

Synonyms Comment Organism
NEP
-
 Homo sapiens

Temperature Optimum [°C]

Temperature Optimum [°C] Temperature Optimum Maximum [°C] Comment Organism
37
-
 assay at Homo sapiens

pH Optimum

pH Optimum Minimum pH Optimum Maximum Comment Organism
6.5
-
 assay at Homo sapiens

Ki Value [mM]

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

General Information Comment Organism
evolution the enzyme is a member of the M13 subgroup of the zinc-dependent endopeptidase family Homo sapiens