3.4.24.56: insulysin
This is an abbreviated version!
For detailed information about insulysin, go to the full flat file.
Word Map on EC 3.4.24.56
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3.4.24.56
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alzheimer
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neprilysin
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abeta
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hippocampus
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dementia
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mellitus
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cerebral
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morris
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amyloid-beta
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metalloprotease
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maze
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neuroprotective
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amyloidogenic
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tau
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beta-amyloid
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neurodegenerative
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presenilin
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endothelin-converting
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senile
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gamma-secretase
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beta-protein
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125i-insulin
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abeta-degrading
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hyperinsulinemia
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metalloendopeptidase
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bacitracin
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beta-secretase
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late-onset
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amylin
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microglia
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medicine
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adam10
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non-amyloidogenic
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glutathione-insulin
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enzyme-1
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b-chains
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analysis
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ad-like
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anti-ide
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exosite
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beta-peptide
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abeta1-40
- 3.4.24.56
- alzheimer
- neprilysin
- abeta
- hippocampus
- dementia
- mellitus
- cerebral
-
morris
- amyloid-beta
- metalloprotease
-
maze
-
neuroprotective
-
amyloidogenic
- tau
- beta-amyloid
- neurodegenerative
-
presenilin
-
endothelin-converting
-
senile
- gamma-secretase
- beta-protein
- 125i-insulin
-
abeta-degrading
- hyperinsulinemia
- metalloendopeptidase
- bacitracin
- beta-secretase
-
late-onset
- amylin
- microglia
- medicine
- adam10
-
non-amyloidogenic
-
glutathione-insulin
- enzyme-1
- b-chains
- analysis
-
ad-like
-
anti-ide
-
exosite
- beta-peptide
- abeta1-40
Reaction
Degradation of insulin, glucagon and other polypeptides. No action on proteins =
Synonyms
ADE, amyloid degrading enzyme, cgd6_5510, EC 3.4.22.11, EC 3.4.99.10, EC 3.4.99.45, gamma-endorphin-generating enzyme, IDE, INS20-19, insulin degrading enzyme, Insulin protease, Insulin proteinase, Insulin-degrading enzyme, Insulin-degrading neutral proteinase, Insulin-glucagon protease, Insulin-specific protease, Insulinase, Insulysin, Metalloinsulinase, More, pitrilysin metallopeptidase 1, Pitrm1
ECTree
Advanced search results
Engineering
Engineering on EC 3.4.24.56 - insulysin
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A140D
specific activity with the substrates (7-methoxycoumarin-4-yl)acetyl-VEALYLVCGEK(2,4-dinitrophenyl)-OH and (7-methoxycoumarin-4-yl)acetyl-QKLVFFAEDVK(2,4-dinitrophenyl)-OH is below 1 nmol/min*mg
A140F
specific activity with the substrates (7-methoxycoumarin-4-yl)acetyl-VEALYLVCGEK(2,4-dinitrophenyl)-OH and (7-methoxycoumarin-4-yl)acetyl-QKLVFFAEDVK(2,4-dinitrophenyl)-OH is below 1 nmol/min*mg
A140K
specific activity with the substrates (7-methoxycoumarin-4-yl)acetyl-VEALYLVCGEK(2,4-dinitrophenyl)-OH and (7-methoxycoumarin-4-yl)acetyl-QKLVFFAEDVK(2,4-dinitrophenyl)-OH is below 1 nmol/min*mg
A140N
specific activity with the substrates (7-methoxycoumarin-4-yl)acetyl-VEALYLVCGEK(2,4-dinitrophenyl)-OH and (7-methoxycoumarin-4-yl)acetyl-QKLVFFAEDVK(2,4-dinitrophenyl)-OH is below 1 nmol/min*mg
A140W
specific activity with the substrates (7-methoxycoumarin-4-yl)acetyl-VEALYLVCGEK(2,4-dinitrophenyl)-OH and (7-methoxycoumarin-4-yl)acetyl-QKLVFFAEDVK(2,4-dinitrophenyl)-OH is below 1 nmol/min*mg
A140Y
specific activity with the substrates (7-methoxycoumarin-4-yl)acetyl-VEALYLVCGEK(2,4-dinitrophenyl)-OH and (7-methoxycoumarin-4-yl)acetyl-QKLVFFAEDVK(2,4-dinitrophenyl)-OH is below 1 nmol/min*mg
C819A
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thiol-directed modification of C819 likely causes local structure perturbation to reduce substrate binding and catalysis
D426C/K899C
E111F
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a mixed dimer in which one subunit contains the wild-type sequence and the other contains a E111F mutation that permits substrate binding, but not catalysis (E111F), exhibits a decrease in turnover number. A mixed dimer consisting of IDE:mutant E111F/Y609F IDE shows a high reduction in kcat, Km (Abz-GGFLRKHGQ-EDDnp) is 66% reduced compared to wild-type. Mixed dimer IDE:mutant E111F in which the inactive subunit can bind substrate exhibites a decreased activity than wild-type IDE towards substrate amyloid beta peptide
E111Q
E341A
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mutant is active in degrading substrate V, relative activity closed to wild-type
E341K
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mutant is active in degrading substrate V, relative activity 15% compared to wild-type. Exosite mutant is unable to further degrade the Ub1-74 fragment
E341Q
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mutant is active in degrading substrate V, relative activity 20% compared to wild-type. Exosite mutant is unable to further degrade the Ub1-74 fragment
F530A
the mutation renders the enzyme hyperactive, with up to a 20fold enhancement in degrading activity
G339P
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mutant is active in degrading substrate V, relative activity 75% compared to wild-type. Exosite mutant is unable to further degrade the Ub1-74 fragment
G361P
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mutant is active in degrading substrate V, relative activity 80% compared to wild-type. Exosite mutant is unable to further degrade the Ub1-74 fragment
H112Q
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a mixed dimer composed of one wild-type subunit and the other subunit containing a H112Q mutation that neither permits substrate binding nor catalysis exhibits the same turnover number per active subunit as wild-type IDE. Mixed oligomer IDE:IDE mutant H112Q shows similar kcat and Km (Abz-GGFLRKHGQ-EDDnp) compared to wild-type. Mixed dimer IDE:mutant H112Q IDE in which the inactive subunit does not bind substrate exhibits a slightly higher activity than wild-type IDE towards substrate amyloid beta peptide
K898A
P286G
the mutation results in a significant reduction of enzyme catalysis
R183a
about 35% of the activity compared to wild-type enzyme with the substrate
R183D
less than 5% of the activity compared to wild-type enzyme with the substrate (7-methoxycoumarin-4-yl)acetyl-KLVFFAEDK(Dnp)-OH
R183E
about less than 5% of the activity compared to wild-type enzyme with the substrate (7-methoxycoumarin-4-yl)acetyl-KLVFFAEDK(Dnp)-OH
R183K
about 30% of the activity compared to wild-type enzyme with the substrate (7-methoxycoumarin-4-yl)acetyl-KLVFFAEDK(Dnp)-OH
R183N
about 10% of the activity compared to wild-type enzyme with the substrate (7-methoxycoumarin-4-yl)acetyl-KLVFFAEDK(Dnp)-OH
R183Q
mutant Pitrm1 R183Q is implicated in inherited amyloidogenic neuropathy. Recombinant R183Q mutant is less active than the recombinant wild-type enzyme against recombinant amyloid beta-peptide (Abeta1-40). R183Q mutant enzyme exhibits significantly decreased rate of fluorogenic peptide hydrolysis ((7-methoxycoumarin-4-yl)acetyl-KLVFFAEDK(Dnp)-OH), yet retains similar binding affinity by comparison with the wild-type enzyme. Residue R183 is positioned within an N-terminal strand-loop-strand motif that is essential for enzyme function. A requirement for charged residues within 4.5 A of residue R183 is demonstrated. The R183Q mutant enzyme exhibits increased sensitivity to heat inactivation
S132C/E817C
Y609F
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mutation Y609F in the distal part of the substrate binding site of the active subunit blocks allosteric activation regardless of the activity of the other subunit. A mixed dimer consisting of mutant Y609F IDE: mutant E111F IDE shows a high reduction in kcat and a reduction in Km compared to wild-type. A mixed dimer consisting of mutant Y609F IDE: mutant E111F/Y609F IDE shows a high reduction in kcat, Km (Abz-GGFLRKHGQ-EDDnp) is 50% reduced compared to wild-type. Substrate amyloid beta peptide: When the distal site is mutated on both subunits (Y609F IDE:IDE Y609F) there is an even greater decrease in the reaction rate
Y831A
specific activity with the substrates (7-methoxycoumarin-4-yl)acetyl-VEALYLVCGEK(2,4-dinitrophenyl)-OH and (7-methoxycoumarin-4-yl)acetyl-QKLVFFAEDVK(2,4-dinitrophenyl)-OH is below 1 nmol/min*mg
Y831D
specific activity with the substrates (7-methoxycoumarin-4-yl)acetyl-VEALYLVCGEK(2,4-dinitrophenyl)-OH and (7-methoxycoumarin-4-yl)acetyl-QKLVFFAEDVK(2,4-dinitrophenyl)-OH is below 1 nmol/min*mg
Y831F
Y831K
specific activity with the substrates (7-methoxycoumarin-4-yl)acetyl-VEALYLVCGEK(2,4-dinitrophenyl)-OH and (7-methoxycoumarin-4-yl)acetyl-QKLVFFAEDVK(2,4-dinitrophenyl)-OH is below 1 nmol/min*mg
Y831N
specific activity with the substrates (7-methoxycoumarin-4-yl)acetyl-VEALYLVCGEK(2,4-dinitrophenyl)-OH and (7-methoxycoumarin-4-yl)acetyl-QKLVFFAEDVK(2,4-dinitrophenyl)-OH is below 1 nmol/min*mg
Y831W
specific activity with the substrates (7-methoxycoumarin-4-yl)acetyl-VEALYLVCGEK(2,4-dinitrophenyl)-OH and (7-methoxycoumarin-4-yl)acetyl-QKLVFFAEDVK(2,4-dinitrophenyl)-OH is below 1 nmol/min*mg
E111A
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change in the substrate response from sigmoidal to hyperbolic. Activating effect of ATP or triphosphate with substrate 2-aminobenzoyl-GGFLRKHGQ-(N-(2,4-dinitrophenyl)ethylenediamine) on wild-type is dampened. Mutant recognizes additional cleavage sites in substrate beta-endorphin
E111F
E111L
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change in the substrate response from sigmoidal to hyperbolic. Activating effect of ATP or triphosphate with substrate 2-aminobenzoyl-GGFLRKHGQ-(N-(2,4-dinitrophenyl)ethylenediamine) on wild-type is dampened. Mutant recognizes additional cleavage sites in substrate beta-endorphin
E111V
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change in the substrate response from sigmoidal to hyperbolic. Activating effect of ATP or triphosphate with substrate 2-aminobenzoyl-GGFLRKHGQ-(N-(2,4-dinitrophenyl)ethylenediamine) on wild-type is dampened. Mutant recognizes additional cleavage sites in substrate beta-endorphin
H112D
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change in the substrate response from sigmoidal to hyperbolic. Activating effect of ATP or triphosphate with substrate 2-aminobenzoyl-GGFLRKHGQ-(N-(2,4-dinitrophenyl)ethylenediamine) on wild-type is changed to inhibition. Affinity to Zn2+ is lower than in wild-type
H112Q
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change in the substrate response from sigmoidal to hyperbolic. Activating effect of ATP or triphosphate with substrate 2-aminobenzoyl-GGFLRKHGQ-(N-(2,4-dinitrophenyl)ethylenediamine) on wild-type is changed to inhibition. Affinity to Zn2+ is lower than in wild-type
H18R/A890V
I374S
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mutation in the distal site eliminates allosterism, Vmax decreased approximately 10fold compared to wild-type, Km (Abz-GGFLRKHGQ-EDDnp) roughly similar to wild-type, no heterotropic activation with bradykinin, no significant activation through ATP
K898A/K899A/S901A
R429S
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mutant shows greatly decreased activation by the polyphosphate anions ATP and PPP. kcat (Abz-GGFLRKHGQ-EDDnp) comparable to wild-type and Km (Abz-GGFLRKHGQ-EDDnp) higher than wild-type
V360S
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mutation in the distal site eliminates allosterism, Vmax decreased approximately 10fold compared to wild-type, Km (Abz-GGFLRKHGQ-EDDnp) roughly similar to wild-type, no heterotropic activation with bradykinin, ATP activation is reduced to 8fold
Y609F
additional information
D426C/K899C
site-directed mutagenesis, hyperactive IDE mutation, the mutant shows increased activity, but reduced activationby ATP compared to the wild-type enzyme
E111Q
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site-directed mutagenesis, the mtant shows highly reduced catalytic activity compared to the wild-typ enzyme
E111Q
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catalysitcally inactive. Mutant forms ordered crystals of considerable size at a more rapid rate than the wild type
E111Q
specific activity with the substrates (7-methoxycoumarin-4-yl)acetyl-VEALYLVCGEK(2,4-dinitrophenyl)-OH and (7-methoxycoumarin-4-yl)acetyl-QKLVFFAEDVK(2,4-dinitrophenyl)-OH is below 1 nmol/min*mg
site-directed mutagenesis, the mutant shows catalytic activity similar to the wild-type enzyme
Y831F
specific activity with the substrates (7-methoxycoumarin-4-yl)acetyl-VEALYLVCGEK(2,4-dinitrophenyl)-OH and (7-methoxycoumarin-4-yl)acetyl-QKLVFFAEDVK(2,4-dinitrophenyl)-OH is below 1 nmol/min*mg
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change in the substrate response from sigmoidal to hyperbolic. Activating effect of ATP or triphosphate with substrate 2-aminobenzoyl-GGFLRKHGQ-(N-(2,4-dinitrophenyl)ethylenediamine) on wild-type is dampened. Mutant recognizes additional cleavage sites in substrate beta-endorphin
E111F
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crystal structure of a catalytically compromised E111F mutant of IDE has electron density for peptide ligands bound at the active site in domain 1 and a distal site in domain 2
plus silent mutation at codon 934, naturally occuring missense mutations occuring in a rat model of type 2 diabetes mellitus, resulting in decreased catalytic efficiency and 15-30% deficit in degradation of both insulin and insulin Abeta. Endogenously secreted insulin Abeta40 and Abeta42 are significantly elevated in primnary neuronal cultures from mutant animals
H18R/A890V
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naturally occuring mutations in the insulysin gene in GK rats causing 30% reduced enzyme activity and type 2 diabetes as well as increased amyloid beta peptide levels , the rats exhibit defects in both insulin action and insulin degradation, the increased amyloid beta-peptide levels do not lead to increased steady-state levels of its activity due to compensatory degradative mechanisms in the brain
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mutant shows greatly decreased activation by the polyphosphate anions ATP and PPP, mutant is also deficient in activation by small peptides and has reduced intracellular function relative to unmodified IDE. Km and kcat (Abz-GGFLRKHGQ-EDDnp) higher than wild-type
K898A/K899A/S901A
variant with mutations in the polyanion-binding site shows reduced activation by myoinositol 1,4,5-trisphosphate and phytic acid
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mutation in the distal site eliminates allosterism, Vmax decreased approximately 10fold compared to wild-type, Km (Abz-GGFLRKHGQ-EDDnp) roughly similar to wild-type, no heterotropic activation with bradykinin, no significant activation through ATP
Y609F
activation by myoinositol 1,4,5-trisphosphate and phytic acid is decreased in the mutant enzyme
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chromosome 10-linked Alzheimer disease families show decreased enzyme activity
additional information
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construction of transgenic mice, termed H1/siRNAinsulin-CMV/hIDE transgenic mice, TG9385, co-expressing specific insulin siRNA sequences and the human insulin degrading enzyme, hIDE, gene resulting in changes in the proteins in the endoplasmic reticulum stress signal pathway and a diabetes-like phenotype with impaired glucose tolerance and lower serum insulin levels compared to the wild-type mice, expression pattern in muscle, lung, brain, liver, kidney, heart, and intestine, tissue-specific regulation, overview
additional information
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enzyme deficiency leads to development of either Alzheimer's disease and type 2 diabetes, overview
additional information
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genotyping of single nucleotide polymorphisms in the IDE gene in Finnish patients with Alzheimer's disease, SNPs rs4646953 and rs4646955 to be associated with Alzheimer's disease, the insulin-degrading enzyme is genetically associated with Alzheimer's disease in the Finnish population, overview
additional information
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human enzyme co-expression with specific insulin siRNA in H1/siRNAinsulin-CMV/hIDE transgenic mice, TG9385, using a chromosome integrated dual-recombinant expression system, in lung, brain, liver, kidney, heart, and intestine, with significantly increased enzyme level and activity in the liver, the transgenic mice show impaired glucose tolerance and reduced serum insulin levels compared to wild-type mice, down-regulation of gene transcripts in the liver of H1/siRNAinsulin-CMV/hIDE mice, gene ontology, overview
additional information
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no correlation of single nucleotide polymorphisms and IDE haplotypes with the risk of dementia, overview
additional information
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overexpression of insulysin in human amyloid precursor protein transgenic mice leads to a decrease in amyloid beta peptide levels
additional information
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siRNA-mediated gene silencing of IDE in Hep-G2 cells leading to 50% reduction of IDE mRNA and protein, short-lived protein degradation is unchanged in the cells with reduced IDE expression, while long-lived and very-long-lived protein degradation is reduced, overview
additional information
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construction of mutants lacking one or severall cysteine resiudes. A mutant devoid of all 13 cysteine residues is insensitive to the inhibition by S-nitrosoglutathione, hydrogen peroxide, or N-ethylmaleimide
additional information
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construction of Cys residue mutants of IDE, properties of oxidized and/or nitrosylated mutant enzymes compared to wild-type enzyme, overview
additional information
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IDE genotyping and identification of genetic variants, the A/T allele of IDE gene variant rs11187033 is associated with the metabolic syndrome, and might contribute to metabolic syndrome susceptibility in Chinese elders, overview
additional information
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IDE genotyping identification of polymorphisms, three polymorphisms occur in IDE promoter: -1002T/G (rs3758505), -179T/C (rs4646953) and -51C/T (rs4646954). The -1002T and -51C alleles are overrepresented in 357 sporadic Alzheimer disease patients when compared to those in 331 healthy individuals. Furthermore, -1002T/G and -51C/T are in strong linkage disequilibrium and they construct a relatively risky -1002T/-51C and a relatively protective -1002G/-51T
additional information
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identification of a genetic variant 311 rs6583817, found in two Croatian isolated populations, unequivocally associated with increased IDE expression that is also associated with reduced plasma amyloidbeta40 and decreased late onset Alzheimer's disease susceptibility. rs6583817 increases reporter gene expression in Be(2)-C and Hep-G2 cell lines. Additional eleven IDE haplotypes, that also show significant association, overview
additional information
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silencing of IDE in Hep-Ge cells by siRNA inhibits insulin degradation by up to 76%
additional information
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Tg3576 transgenic mice express the Swedish mutant of amyloid beta-peptide, the expression and activity of IDE in the transgenic mice is increased with age around plaques as a component of astrocyte activation due to amyloid beta-peptide-triggered inflammation in contrast to the behaviour in Alzheimer mice, overview
additional information
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IDE knockout mice show decreased insulin degradation and associated hyperinsulinemia
additional information
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heterologous expression in Saccharomyces cerevisiae, enzyme can promote the in vivo production of yeast a-factor mating pheromone, but cannot substitue for other fuctions of yeast Axl1p. Enzyme requires an intact C-terminus for optimal activity
additional information
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construction of transgenic mice with increased enzyme levels show that the enzyme prevents and treats Alzheimer's disease formation
additional information
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mutant with a deleted putative dimer interface in the C-terminal region is created, which results in a monomeric variant. Monomeric IDE retains enzymatic activity. Monomeric IDE retains, 25% of the wild type activity substrate (Abz-GGFLRKHGQ-EDDnp). With the peptide substrates beta-endorphin and amyloid beta peptide 1-40, monomeric IDE retains 1 to 0.25% of wild type activity. No activation through bradykinin or dynorphin B-9. Monomeric IDE is not activated by polyphosphates