A high molecular weight metalloendoprotease from the cytosol of mammalian cells.

Kirschner, Richard J.; Goldberg, Alfred L.

doi:10.1016/s0021-9258(18)33145-4

Cited by 89 publications

(6 citation statements)

References 53 publications

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“…H uman insulin-degrading enzyme (IDE, EC 3.4.24.56, GenBank reference sequence BC096336) is a 110 kDa chambered zinc metalloendopeptidase of the M16A subfamily. The enzyme is notable for its role in the proteolysis of several physiologically significant peptides that adopt β-structures, including insulin, 1,2 glucagon, 3 amylin, 4 and amyloid β (Aβ) peptides. 5 Multiple studies point to roles for IDE in metabolic signaling and disease.…”

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confidence: 99%

Molecular Determinants of Substrate Specificity in Human Insulin-Degrading Enzyme

et al. 2018

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Insulin-degrading enzyme (IDE) is a 110 kDa chambered zinc metalloendopeptidase that degrades insulin, amyloid β, and other intermediate-sized aggregation prone peptides that adopt β-structures. Structural studies of IDE in complex with multiple physiological substrates have suggested a role for hydrophobic and aromatic residues of the IDE active site in substrate binding and catalysis. Here, we examine functional requirements for conserved hydrophobic and aromatic IDE active site residues that are positioned within 4.5 Å of IDE-bound insulin B chain and amyloid β peptides in the reported crystal structures for the respective enzyme-substrate complexes. Charge, size, hydrophobicity, aromaticity, and other functional group requirements for substrate binding IDE active site residues were examined through mutational analysis of the recombinant human enzyme and enzyme kinetic studies conducted using native and fluorogenic derivatives of human insulin and amyloid β peptides. A functional requirement for IDE active site residues F115, A140, F141, Y150, W199, F202, F820, and Y831 was established, and specific contributions of residue charge, size, and hydrophobicity to substrate binding, specificity, and proteolysis were demonstrated. IDE mutant alleles that exhibited enhanced or diminished proteolytic activity toward insulin or amyloid β peptides and derivative substrates were identified.

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confidence: 99%

Molecular Determinants of Substrate Specificity in Human Insulin-Degrading Enzyme

et al. 2018

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“…The mean duration of disease of PD patients was 5.55 ± 3.30 years (1)(2)(3)(4)(5)(6)(7)(8)(9)(10)(11)(12), and the age of onset of disease was 63.11 ± 9.83 (37-85) years. The initial symptoms of the patients were tremor at a rate of 63.6%, bradykinesia at a rate of 31.8%, and postural instability at a rate of 4.5%.…”

Section: Resultsmentioning

confidence: 99%

Is there a role of insulin degrading enzyme in Parkinson’s disease?

Aktürk¹,

Çakır²,

Saçmacı³

et al. 2022

NeuroAsia

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Objectives: Alpha-synucleins are the basic structural components of Lewy bodies, which are the pathognomonic feature of Parkinson’s disease (PD). It is propounded that alpha-synucleins interact with the insulin-degrading enzyme (IDE). In this study, it was aimed to evaluate the relationship between the insulin-degrading enzyme (IDE) level and the disease and the symptomatology of the disease in patients with PD. Methods: A total of 98 individuals, including 57 patients with PD and 41 healthy controls, were enrolled in this cross-sectional, case-control study. The clinical characteristics of the patients with PD, modified Hoehn and Yahr (mHY) stages, and Unified Parkinson’s Disease Rating Scale (UPDRS) scores were recorded. The mini-mental state examination (MMSE) was applied to all participants. Serum samples were collected for the level of IDE. Results: No significant difference was found in terms of IDE between PD patients and the control group with similar sociodemographic characteristics (p>0.05). No statistically significant difference was revealed between the level of IDE and the age of onset of disease, initial symptom, disease subtype, mHY stage, LED dose, MMSE score, UPDRS part II, III and IV. In PD patients, a weak negative correlation was found between IDE, duration of disease, and UPDRS part I (p=0.049, r=-0.299 and p=0.003, r=-0.431, respectively). Conclusion: No significant difference was determined in the level of IDE between PD patients and the controls. There was no relationship between IDE level and onset symptom, disease subtype, disease stage. It was suggested that the serum level of IDE was related to a longer duration of disease and the affected mentation, behavior and mood. The confirmation of these results with larger patient series will contribute to clarifying the role of IDE in the pathogenesis of PD.

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“…The molecular weight of ghrelin is ~3400 kDa. Consistent with peptide degradation by a zinc-dependent metallopeptidase, disappearance of a low MW fragment in the presence of catalytically active IDE was sensitive to and limited by pretreatment of the enzymatic activity with ethylenediaminetetraacetate (EDTA), a metal chelating agent and established IDE inhibitor [Kirschner and Goldberg 1983].…”

Section: Resultsmentioning

confidence: 99%

“…Ghrelin degradation assays were performed after separately assembling pre-mixtures containing 30 ng/μL IDE, 30 ng/μL neprilysin or 100 ng/μL GST, and 0.5 μg/μL substrate (octanoylated ghrelin). Where present for purposes of enzyme inhibition prior to the proteolytic experiment, 500 mM EDTA, an established IDE inhibitor [Kirschner and Goldberg 1983], was added to the enzyme pre-mixture during initial assembly. Enzyme and substrate pre-mixtures were pre-incubated at assay temperature of 37 °C, and the assay was initiated upon mixing of enzyme and substrate.…”

Section: Methodsmentioning

confidence: 99%

“…Proteolytic reactions were quenched at endpoint by addition of SDS-PAGE sample buffer containing 160 mM Tris, pH 6.8, 100 mM EDTA, 32% w/v glycerol, 1.25% w/v β-mercaptoethanol and 0.003% w/v bromophenol blue. Where present for purposes of enzyme inhibition, EDTA, an established IDE inhibitor [Kirschner and Goldberg 1983], was added to the enzyme pre-mixture during assembly. Samples were separated by SDS-PAGE (BioRad Any kD ™ Mini-PROTEAN ® TGX gel, manufacturer’s Tris-glycine buffer system) immediately after the proteolytic assay.…”

Section: Methodsmentioning

confidence: 99%

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Ghrelin proteolysis by insulin-degrading enzyme

et al. 2021

Preprint

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Here we report proteolysis of synthetic acylated human ghrelin by recombinant human insulin-degrading enzyme (IDE). Kinetic parameters and sites of proteolytic cleavage were evaluated. Ghrelin proteolysis by IDE was inhibited by ethylenediaminetetraacetate (EDTA), a metal chelating agent. Ghrelin proteolysis appears at least somewhat specific to M16 family proteases such as IDE, as the M13 protease neprilysin (NEP) did not exhibit ghrelin proteolysis in this study. A quenched fluorogenic peptide substrate comprising the primary sites of IDE-mediated ghrelin proteolysis (Mca-QRVQQRKESKK(Dnp)-OH; Mca: 7-methoxycoumarin-3-carboxylic acid; Dnp: 2,4-dinitrophenyl) was developed and used to evaluate enzyme specificity and kinetic parameters of proteolysis. Like acyl ghrelin, Mca-QRVQQRKESKK(Dnp)-OH was efficiently cleaved by IDE central to the target sequence. We anticipate that this quenched fluorogenic peptide substrate will be of value to future studies of ghrelin proteolysis by IDE and potentially other peptidases.

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A high molecular weight metalloendoprotease from the cytosol of mammalian cells.

Cited by 89 publications

References 53 publications

Molecular Determinants of Substrate Specificity in Human Insulin-Degrading Enzyme

Molecular Determinants of Substrate Specificity in Human Insulin-Degrading Enzyme

Is there a role of insulin degrading enzyme in Parkinson’s disease?

Ghrelin proteolysis by insulin-degrading enzyme

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