Insulin-Degrading Enzyme

Duckworth, William C.

doi:10.1007/978-3-642-74098-5_8

Cited by 9 publications

(9 citation statements)

References 97 publications

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“…The molecular weight of IDE identified is consistent with the reported molecular weight Bai et al 1995b). Interestingly, the molecular weight of IDE in rat intestine is similar to that in human intestine (Duckworth 1990).…”

Section: Inimunoprecipitation Und Western Blotsmentioning

confidence: 72%

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The Presence of Insulin-degrading Enzyme in Human Ileal and Colonic Mucosal Cells

Bai

Hong

Enberger

et al. 1996

Journal of Pharmacy and Pharmacology

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The aim of this research is to characterize the presence of insulin-degrading enzyme in human colon and ileal mucosal cells. Biochemical studies, including the activity-pH profiles, the effects of enzyme inhibitors, immunoprecipitation and western blots, were conducted. The majority of insulin-degrading activity in colon mucosal cells was localized in the cytosol. In both colon and ileum, cytosolic insulin-degrading activities had a pH optimum at pH 7.5, and were extensively inhibited by each of N-ethylmaleimide, p-chloromercuribenzoate, and 1,10-phenanthroline, but were very weakly affected by each of leupeptin, chymostatin, diisopropyl phosphofluoridate and soybean trypsin inhibitor. In the colon and ileum, more than 93% and 96%, respectively, of cytosolic insulin-degrading activities were removed by the mouse monoclonal antibody to human RBC insulin-degrading enzyme, as compared with less than 20% by the normal mouse IgG for both tissues. Further, a western blot analysis revealed that a cytosolic protein of 110 kD, in both human colon and ileum, reacted with the monoclonal antibody to insulin-degrading enzyme. It is concluded that insulin-degrading enzyme is present in the cytosol of human colon and ileal mucosal cells.

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Section: Inimunoprecipitation Und Western Blotsmentioning

confidence: 72%

“…The final TCA concentration and pH in the mixture were 7.8% and less than I , respectively. In general, a final TCA concentration below 10% was used (Duckworth 1990). The resulting mixture was then centrifuged at 6000 g for 10 min.…”

Section: Insulin Degradation In Subcellular Fractionsmentioning

confidence: 99%

The Presence of Insulin-degrading Enzyme in Human Ileal and Colonic Mucosal Cells

Bai

Hong

Enberger

et al. 1996

Journal of Pharmacy and Pharmacology

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“…The apparent Michaelis-Menten constant (K,) for alveolar epithelial cell cytosol was 135nM (Figure 2), larger than the 20-40nM for purified IDE (Duckworth 1990). The K, value reported for crude tissues was 2 9 0 m for mouse pancreatic acini (Goldfine et a1 1984), 140 mM for cultured rat heDatoma cells (Harada et a1 1993) high molecular-weight non-lysosomal protease, has recently been reported to have trypsin-, chymotrypsin-and cucumsin-like activity and it is therefore possible that it is involved in insulin metabolism (Rivett 1989;Orlowski 1990).…”

Section: Resultsmentioning

confidence: 88%

“…Insulin-degrading enzyme (IDE) (EC 3.4.22.1 1) has been implicated in intracellular degradation of insulin (Hamel et a1 1987;Duckworth 1988). IDE accounts for most of the insulin-degrading activity in extracts of muscle, liver, kidney, fat cells, erythrocytes, fibroblasts, placenta and pancreas (Stentz et a1 1985;Shii & Roth 1986;Duckworth 1990) with activity mostly in the cytoplasm (Duckworth et a1 1972). IDE is inhibited by metal chelators (ethylenediaminetetraacetic acid, ethyleneglycolbis-(fi-aminoethyl ether), N,N'-tetraacetic acid and 1,lO-phenanthroline) and sulphhydryl inhibitors (N-ethylmaleimide and p-chloromercuribenzoic acid) (Duckworth 1990).…”

mentioning

confidence: 99%

Investigation into the Presence of Insulin-degrading Enzyme in Cultured Type I1 Alveolar Cells and the Effects of Enzyme Inhibitors on Pulmonary Bioavailability of Insulin in Rats

Hsu

Bai

1998

Journal of Pharmacy and Pharmacology

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The purpose of this study was to investigate the role of insulin-degrading enzyme (IDE, EC 3.4.22.11) in insulin degradation in alveolar epithelium. The primary culture of isolated rat type-II pneumocytes was used for the in-vitro characterization of IDE. Insulin was then administered intratracheally with various inhibitors to assess the improvement in its pulmonary bioavailability. In cultured type-II pneumocytes, the cytosolic insulin-degrading activity contributed 81% of total insulin degradation, reached a maximum at pH 7.5 and had an apparent Michaelis-Menten constant (Km) of 135 nM. N-Ethylmaleimide, p-chloromercuribenzoic acid and 1,10-phenanthroline inhibited insulin-degrading activity almost completely in both crude homogenate and cytosol. An immunoprecipitation study showed that IDE contributed 74% of cytosolic insulin-degrading activity. Western blot analysis showing a single band of 110 kDa on reduced SDS (sodium dodecylsulphate) gels confirmed the presence of IDE in cultured type-II cells. When given intratracheally with insulin, inhibitors including N-ethylmaleimide, p-chloromercuribenzoic acid, and 1,10-phenanthroline significantly enhanced the absolute bioavailability of insulin and the compound's hypoglycaemic effects. These results suggest that IDE is present in alveolar epithelium and might be involved in limiting insulin absorption in the lung.

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“…It belongs to the M16 (pitrilysin) family of zinc-metalloendopeptidases, namely inverzincins, characterized by the inverted zinc binding motif HXXEH (2). IDE is present in human beings, animals, fungi, and plants (2-4; http://merops.sanger.ac.uk/, see distribution within family M16.002: insulysin) and is reported to be expressed in the liver, adipocytes, muscle cells, erythrocytes, and kidney (3)(4)(5) but also in cell types not responsive to insulin (6) (www.genecards.org/cgi-bin/carddisp.pl?gene=IDE). Despite its predominant presence in the cytosol, IDE is also found in small but significant amounts in subcellular compartments such as the plasma membrane, endosomes, peroxisomes, and mitochondria (7)(8)(9)(10)(11)(12)(13)(14)(15).…”

Section: Introductionmentioning

confidence: 99%

Expression of metalloprotease insulin-degrading enzyme insulysin in normal and malignant human tissues

Yfanti

Mengele²,

Gkazepis³

et al. 1998

Int J Mol Med

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Abstract. Insulin-degrading enzyme (IDE, insulysin, insulinase; EC 3.4.22.11), a thiol metalloendopeptidase, is involved in intracellular degradation of insulin, thereby inhibiting its translocation and accumulation to the nucleus. Recently, protein expression of IDE has been demonstrated in the epithelial ducts of normal breast and breast cancer tissue. Utilizing four different antibodies generated against different epitopes of the IDE molecule, we performed Western blot analysis and immunohistochemical staining on several normal human tissues, on a plethora of tumor cell lines of different tissue origin, and on malignant breast and ovarian tissue. Applying the four IDE-directed antibodies, we demonstrated IDE expression at the protein level, by means of immunoblotting and immunocytochemistry, in each of the tumor cell lines analyzed. Insulin-degrading enzyme protein expression was found in normal tissues of the kidney, liver, lung, brain, breast and skeletal muscle, as well as in breast and ovarian cancer tissues. Immunohistochemical visualization of IDE indicated cytoplasmic localization of IDE in each of the cell lines and tissues assessed. In conclusion, we performed for the first time a wide-ranging survey on IDE protein expression in normal and malignant tissues and cells thus extending our knowledge on the cellular and tissue distribution of IDE, an enzyme which to date has mainly been studied in connection with Alzheimer's disease and diabetes but not in cancer.

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Insulin-Degrading Enzyme

Cited by 9 publications

References 97 publications

The Presence of Insulin-degrading Enzyme in Human Ileal and Colonic Mucosal Cells

The Presence of Insulin-degrading Enzyme in Human Ileal and Colonic Mucosal Cells

Investigation into the Presence of Insulin-degrading Enzyme in Cultured Type I1 Alveolar Cells and the Effects of Enzyme Inhibitors on Pulmonary Bioavailability of Insulin in Rats

Expression of metalloprotease insulin-degrading enzyme insulysin in normal and malignant human tissues

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