Site directed mutagenesis of insulin-degrading enzyme allows singling out the molecular basis of peptidase <i>versus</i> E1-like activity: the role of metal ions

Bellia, Francesco; Lanza, Valeria; Ahmed, Ikhlas Mohamed Mohamud; García‐Viñuales, Sara; Veiss, Eva; Arizzi, Mariaconcetta; Calcagno, Damiano; Milardi, Danilo; Grasso, Giuseppe

doi:10.1039/c8mt00288f

Cited by 11 publications

(11 citation statements)

References 29 publications

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“…These results demonstrate that Ins dyshomeostasis due to the type 2 or type 1 diabetes can have a direct impact on pain transmission and pain threshold through an IDE mediated mechanism. This result is in line with all the recent findings, which demonstrate a multifaceted role for this enzyme in the brain [14,15,16,31].…”

Section: Discussionsupporting

confidence: 93%

“…In the case of OFQ/N, only very few studies investigate the metabolic processes undergone by the peptide and its fragments [13]. In this work, we report that insulin-degrading enzyme (IDE), an enzyme known for its multifaceted roles in cells [14,15,16,17] and its capability to degrade several different peptides [18,19,20,21], is capable of degrading OFQ/N in vitro, producing a characteristic proteolytic pattern. As IDE is present in the brain [22], which also contains Ins pools, it cannot be excluded that this enzyme indirectly participates in neural communication of pain signals and that neuropeptides involved in pain transmission may contribute to the regulation of IDE activity.…”

Section: Introductionmentioning

confidence: 99%

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IDE Degrades Nociceptin/Orphanin FQ through an Insulin Regulated Mechanism

Zingale

Bellia

Ahmed

et al. 2019

IJMS

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Insulin-degrading enzyme (IDE) was applied to catalyze hydrolysis of Nociceptin/Orphanin 1-16 (OFQ/N) to show the involvement of the enzyme in degradation of neuropeptides engaged in pain transmission. Moreover, IDE degradative action towards insulin (Ins) was inhibited by the OFQ/N fragments, suggesting a possible regulatory mechanism in the central nervous system. It has been found that OFQ/N and Ins affect each other degradation by IDE, although in a different manner. Indeed, while the digestion of OFQ/N is significantly affected by the presence of Ins, the kinetic profile of the Ins hydrolysis is not affected by the presence of OFQ/N. However, the main hydrolytic fragments of OFQ/N produced by IDE exert inhibitory activity towards the IDE-mediated Ins degradation. Here, we present the results indicating that, besides Ins, IDE cleaves neuropeptides and their released fragments act as inhibitors of IDE activity toward Ins. Having in mind that IDE is present in the brain, which also contains Ins receptors, it cannot be excluded that this enzyme indirectly participates in neural communication of pain signals and that neuropeptides involved in pain transmission may contribute to the regulation of IDE activity. Finally, preliminary results on the metabolism of OFQ/N, carried out in the rat spinal cord homogenate in the presence of various inhibitors specific for different classes of proteases, show that OFQ/N proteolysis in rat spinal cord could be due, besides IDE, also to a cysteine protease not yet identified.

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Section: Discussionsupporting

confidence: 93%

Section: Introductionmentioning

confidence: 99%

IDE Degrades Nociceptin/Orphanin FQ through an Insulin Regulated Mechanism

Zingale

Bellia

Ahmed

et al. 2019

IJMS

Self Cite

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“…The final volume was fivefold diluted with an aqueous solution containing 5 % acetonitrile and 0.05 % trifluoroacetic acid before spectrometric analysis by means of MALDI‐MS. This MALDI‐MS analysis was performed by using a protocol already reported [89] . To structurally characterize the ACR adducts, precursor selection for MS/MS analysis was made by using the following criteria: minimum signal‐to‐noise (S/N) ratio, 20; precursor mass tolerance between spots, ±200 ppm.…”

Section: Methodsmentioning

confidence: 99%

“…This MALDI-MS analysis was performed by using ap rotocol already reported. [89] To structurally characterize the ACR adducts, precursor selection for MS/MS analysis was made by using the following criteria:m inimum signal-to-noise (S/N) ratio, 20;p recursor mass tolerance between spots, AE 200 ppm. All MS/MS spectra were acquired by using collision induced dissociation (CID), with 1kVc ollision energy and air as the collision gas, by an accumulation of 5000 laser shots.…”

Section: Teac Assaymentioning

confidence: 99%

Carnoquinolines Target Copper Dyshomeostasis, Aberrant Protein–Protein Interactions, and Oxidative Stress

Bellia

Grasso

Ahmed

et al. 2020

Chemistry A European J

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Metal dysregulation, oxidative stress, protein modification, and aggregation are factors strictly interrelated and associated with neurodegenerative pathologies. As such, all of these aspects represent valid targets to counteract neurodegeneration and, therefore, the development of metal‐binding compounds with other properties to combat multifactorial disorders is definitely on the rise. Herein, the synthesis and in‐depth analysis of the first hybrids of carnosine and 8‐hydroxyquinoline, carnoquinolines (CarHQs), which combine the properties of the dipeptide with those of 8‐hydroxyquinoline, are reported. CarHQs and their copper complexes were characterized through several techniques, such as ESI‐MS and NMR, UV/Vis, and circular dichroism spectroscopy. CarHQs can modulate self‐ and copper‐induced amyloid‐β aggregation. These hybrids combine the antioxidant activity of their parent compounds. Therefore, they can simultaneously scavenge free radicals and reactive carbonyl species, thanks to the phenolic group and imidazole ring. These results indicate that CarHQs are promising multifunctional candidates for neurodegenerative disorders and they are worthy of further studies.

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“…As a result, the Zn(II) binding site is closed, which is irreversibly inactivated [375]. Additionally, the proteolytic activity of IDE was inhibited by Cu(II); however, ubiquitin-activating (E1-like) activity was not affected up to 20 µM [376].…”

Section: Redox-active Metal Ions With Adementioning

confidence: 99%

Redox-Active Metal Ions and Amyloid-Degrading Enzymes in Alzheimer’s Disease

Kim

Lee

2021

IJMS

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Redox-active metal ions, Cu(I/II) and Fe(II/III), are essential biological molecules for the normal functioning of the brain, including oxidative metabolism, synaptic plasticity, myelination, and generation of neurotransmitters. Dyshomeostasis of these redox-active metal ions in the brain could cause Alzheimer’s disease (AD). Thus, regulating the levels of Cu(I/II) and Fe(II/III) is necessary for normal brain function. To control the amounts of metal ions in the brain and understand the involvement of Cu(I/II) and Fe(II/III) in the pathogenesis of AD, many chemical agents have been developed. In addition, since toxic aggregates of amyloid-β (Aβ) have been proposed as one of the major causes of the disease, the mechanism of clearing Aβ is also required to be investigated to reveal the etiology of AD clearly. Multiple metalloenzymes (e.g., neprilysin, insulin-degrading enzyme, and ADAM10) have been reported to have an important role in the degradation of Aβ in the brain. These amyloid degrading enzymes (ADE) could interact with redox-active metal ions and affect the pathogenesis of AD. In this review, we introduce and summarize the roles, distributions, and transportations of Cu(I/II) and Fe(II/III), along with previously invented chelators, and the structures and functions of ADE in the brain, as well as their interrelationships.

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Site directed mutagenesis of insulin-degrading enzyme allows singling out the molecular basis of peptidase versus E1-like activity: the role of metal ions

Abstract: Four specifically designed IDE mutants have been used to unveil the molecular basis of peptidase versus E1-like activity of the enzyme.

Cited by 11 publications

References 29 publications

IDE Degrades Nociceptin/Orphanin FQ through an Insulin Regulated Mechanism

IDE Degrades Nociceptin/Orphanin FQ through an Insulin Regulated Mechanism

Carnoquinolines Target Copper Dyshomeostasis, Aberrant Protein–Protein Interactions, and Oxidative Stress

Redox-Active Metal Ions and Amyloid-Degrading Enzymes in Alzheimer’s Disease

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