Insulin is degraded extracellularly in wounds  by insulin-degrading enzyme (EC 3.4.24.56)

The exact mechanism by which insulin reverses impaired wound healing is unknown. Previous investigators have shown that insulin is degraded in experimental wounds, suggesting that the action of insulin may be locally modified. The following study corroborates these findings and identifies the major proteinase responsible for insulin degradation in wound fluid (WF). Adult male Fisher rats were wounded by subcutaneous implantation of polyvinyl alcohol sponges while under pentobarbital sodium anesthesia. WF and serum were collected on 1, 5, 10, and 14 days postinjury. Decreased insulin concentration in late WF correlated with an increased insulin-degrading activity. Multiple proteinases appear to participate in the overall degradation of insulin in WF. However, the primary enzyme responsible for insulin degradation in WF was characterized by immunoprecipitation and immunoblotting and identified as the neutral thiol-dependent metalloproteinase, insulin-degrading enzyme (EC 3.4.24.56 ). Exogenous steroid administration caused a decrease in WF insulin-degrading activity. Glucagon and adrenocorticotrophin degradation was also observed, whereas minimal degradation of insulin-like growth factors I and II and epidermal growth factor was detected in WF. The ability to extracellularly degrade insulin may represent a unique mechanism for the regulation of this hormone’s role in healing wounds.

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Inhibition of Insulin Degrading Enzyme and Insulin Degradation by UV-Killed Lactobacillus acidophilus

Medical Sciences ◽

10.3390/medsci6020036 ◽

2018 ◽

Vol 6 (2) ◽

pp. 36

Author(s):

Nadia Neyazi ◽

Elahe Motevaseli ◽

Mohammad Khorramizadeh ◽

Taiebeh Mohammadi Farsani ◽

Zahra Nouri ◽

...

Keyword(s):

Lactobacillus Acidophilus ◽

Insulin Degradation ◽

Insulin Degrading Enzyme ◽

Degrading Enzyme

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An insulin epidermal growth factor-binding protein from Drosophila has insulin-degrading activity.

The Journal of Cell Biology ◽

10.1083/jcb.108.1.177 ◽

1989 ◽

Vol 108 (1) ◽

pp. 177-182 ◽

Cited By ~ 8

Author(s):

J V Garcia ◽

M P Stoppelli ◽

S J Decker ◽

M R Rosner

Keyword(s):

Growth Factor ◽

Egf Receptor ◽

Transforming Growth Factor ◽

Binding Protein ◽

Polyclonal Antiserum ◽

Insulin Degrading Enzyme ◽

Developmentally Regulated ◽

Degrading Enzyme ◽

Epidermal Growth ◽

Factor Alpha

We have recently described the purification and characterization of an insulin-degrading enzyme (IDE) from Drosophila melanogaster that can cleave porcine insulin, is highly conserved through evolution and is developmentally regulated. We now report that the IDE is, in fact, an insulin EGF-binding protein (dp100) that we had isolated previously from Drosophila using an antihuman EGF receptor antiserum. This conclusion is based upon the following evidence. (a) dp100, identified by its ability to cross-link to labeled insulin, EGF, and transforming growth factor-alpha (TGF-alpha), and to be immunoprecipitated by anti-EGF receptor antisera, copurifies with the IDE activity. Thus, the purified IDE can be affinity labeled with either 125I-insulin, 125I-EGF, or 125I-TGF-alpha, and this labeling is specifically inhibited with unlabeled insulin, EGF, and the insulin B chain. (b) The antiserum to the human EGF receptor, which recognizes dp100, is able to specifically immunoprecipitate the insulin-degrading activity. (c) The purified IDE preparation contains a single protein of 110 kD which is recognized by both the anti-EGF receptor antiserum and anti-Drosophila IDE antiserum. (d) Polyclonal antiserum to the purified IDE, which specifically recognized only the 110-kD band in Drosophila Kc cells, immunoprecipitates dp100 cross-linked to 125I-TGF-alpha and dp100 cross-linked to 125I-insulin from the purified IDE preparation. (e) EGF, which competes with insulin for binding to dp100, also inhibits the degradation of insulin by the purified IDE. These results raise the possibility that a functional interaction between the insulin and EGF growth factor families can occur which is mediated by the insulin-degrading enzyme.

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ATP Inhibits Insulin-Degrading Enzyme Activity

Experimental Biology and Medicine ◽

10.1177/153537020122600411 ◽

2001 ◽

Vol 226 (4) ◽

pp. 334-341 ◽

Cited By ~ 36

Author(s):

M.C. Camberos ◽

A.A. Pérez ◽

D.P. Udrisar ◽

M.I. Wanderley ◽

J.C. Cresto

Keyword(s):

Enzyme Inhibitors ◽

Inhibitory Effect ◽

Hill Coefficient ◽

Insulin Degradation ◽

Insulin Degrading Enzyme ◽

Degrading Enzyme ◽

Dependent Inhibition ◽

Degradation Activity ◽

Insulin Degrading Enzyme Activity

We studied the ability of ATP to inhibit in vitro the degrading activity of insulin-degrading enzyme. The enzyme was purified from rat skeletal muscle by successive chromatographic steps. The last purification step showed two bands at 110 and 60 kDa in polyacrylamide gel. The enzyme was characterized by its insulin degradation activity, the substrate competition of unlabeled to labeled insulin, the profile of enzyme inhibitors, and the recognition by a specific antibody. One to 5 mM ATP induced a dose-dependent inhibition of insulin degradation (determined by trichloroacetic acid precipitation and insulin antibody binding). Inhibition by 3 mM adenosine 5′-diphosphate, adenosine 5′-monophosphate, guanosine 5′-triphosphate, pyrophosphate, β-γ-methyleneadenosine 5′-triphosphate, adenosine 5′-O-(3 thiotriphosphate), and dibutiryl cyclic adenosine 5′-monophosphate was 74%, 4%, 38%, 46%, 65%, 36%, and 0%, respectively, of that produced by 3 mM ATP. Kinetic analysis of ATP inhibition suggested an allosteric effect as the plot of 1/v (insulin degradation) versus ATP concentration was not linear and the Hill coefficient was more than 1 (1.51 and 2.44). The binding constant for allosteric inhibition was K1T = 1.5 × 10–7 M showing a decrease of enzyme affinity induced by ATP. We conclude that ATP has an inhibitory effect on the insulin degradation activity of the enzyme.

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Targeting Insulin-Degrading Enzyme in Insulin Clearance

International Journal of Molecular Sciences ◽

10.3390/ijms22052235 ◽

2021 ◽

Vol 22 (5) ◽

pp. 2235 ◽

Cited By ~ 1

Author(s):

Malcolm A. Leissring ◽

Carlos M. González-Casimiro ◽

Beatriz Merino ◽

Caitlin N. Suire ◽

Germán Perdomo

Keyword(s):

Diabetes Mellitus ◽

Insulin Clearance ◽

Diabetic Patients ◽

Insulin Degradation ◽

Insulin Degrading Enzyme ◽

Degrading Enzyme ◽

Intermediate Size ◽

Pros And Cons ◽

Hepatic Insulin Clearance

Hepatic insulin clearance, a physiological process that in response to nutritional cues clears ~50–80% of circulating insulin, is emerging as an important factor in our understanding of the pathogenesis of type 2 diabetes mellitus (T2DM). Insulin-degrading enzyme (IDE) is a highly conserved Zn2+-metalloprotease that degrades insulin and several other intermediate-size peptides. Both, insulin clearance and IDE activity are reduced in diabetic patients, albeit the cause-effect relationship in humans remains unproven. Because historically IDE has been proposed as the main enzyme involved in insulin degradation, efforts in the development of IDE inhibitors as therapeutics in diabetic patients has attracted attention during the last decades. In this review, we retrace the path from Mirsky’s seminal discovery of IDE to the present, highlighting the pros and cons of the development of IDE inhibitors as a pharmacological approach to treating diabetic patients.

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Androgen- and Estrogen-Dependent Regulation of Insulin-Degrading Enzyme in Subcellular Fractions of Rat Prostate and Uterus

Experimental Biology and Medicine ◽

10.1177/153537020523000706 ◽

2005 ◽

Vol 230 (7) ◽

pp. 479-486 ◽

Cited By ~ 18

Author(s):

Daniel P. Udrisar ◽

Maria I. Wanderley ◽

Regina C. C. Porto ◽

Carla L. P. Cardoso ◽

Maria C. L. Barbosa ◽

...

Keyword(s):

Estrous Cycle ◽

Cytosolic Fraction ◽

Male Rat ◽

Insulin Degradation ◽

Nuclear Fraction ◽

Female Rat ◽

Insulin Degrading Enzyme ◽

Degrading Enzyme ◽

Uterine Growth ◽

Rat Prostate

Innumerous data support the fact that insulin-degrading enzyme (IDE) is the primary enzymatic mechanism for initiating and controlling cellular insulin degradation. Nevertheless, insulin degradation is unlikely to be the only cellular function of IDE, because it appears that some cellular effects of insulin are mediated by IDE as a regulatory protein. Insulin-degrading enzyme shows a significant correlation with various cellular functions, such as cellular growth and differentiation, and the expression of IDE is developmentally regulated. Besides insulin, other substrates are also degraded by IDE, including various growth-promoting peptides. It has also been shown that IDE enhances the binding of androgen to DNA in the nuclear compartment. It is also known that the androgen hormones have a stimulatory effect on prostate growth, and that estradiol stimulates uterine growth. To establish whether IDE is regulated by a cellular prostate/uterine growth stimulus, the present study assessed whether IDE was modified in quantity and activity during proliferative conditions (castration + testosterone in the male rat, or castration + estradiol or the proestrus phase of the estrous cycle in the female rat) and autolysis (castration or the metestrus phase of the estrous cycle) using cytosolic and nuclear fractions of rat prostate and cytosolic fractions of rat uterus. The activity and amount of IDE decreased in the cytosolic fraction with castration and during metestrus, and increased with testosterone or estradiol treatment and during proestrus. In the nuclear fraction, the quantity of the IDE followed the same pattern observed in the cytosolic fraction, although without degradative activity. The data presented here suggest that IDE may participate in prostatic and uterine growth and that the testosterone or estradiol and/or prostate and uterus insulin-like growth factors may be important factors for the expression and regulation of IDE in the prostate and uterus

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