• Title/Summary/Keyword: degradation stabilization

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Dose-dependent UV Stabilization of p53 in Cultured Human Cells Undergoing Apoptosis Is Mediated by Poly(ADP-ribosyl)ation

  • Won, Jungyeon;Chung, So Young;Kim, Seung Beom;Byun, Boo Hyeong;Yoon, Yoo Sik;Joe, Cheol O.
    • Molecules and Cells
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    • v.21 no.2
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    • pp.218-223
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    • 2006
  • The effect of poly(ADP-ribosyl)ation on the stability of p53 in SK-HEP1 cells treated with UV light was examined. Intracellular levels of p53 increased in cells treated with a low dose of UV light ($20J/m^2$), whereas they increased but then declined after a higher dose of UV ($100J/m^2$). Intracellular levels of p53 in the UV treated SK-HEP1 cells were dependent on the UV dose. Use of proteasome inhibitors revealed that p53 is degraded by proteasomal proteolysis after high doses of UV light. We present evidence that, at low doses, poly(ADP-ribose)polymerase (PARP) poly(ADP-ribosyl) ates p53 and protects it from proteasomal degradation before caspase-3 is activated, whereas at high doses the cells undergo UV induced apoptosis and PARP is cleaved by caspase-3 before it can protect p53 from degradation. Destabilization of p53 by cleavage of PARP may be important in cell fate decision favoring apoptosis.

MS2 Labeling of Endogenous Beta-Actin mRNA Does Not Result in Stabilization of Degradation Intermediates

  • Kim, Songhee H.;Vieira, Melissa;Kim, Hye-Jin;Kesawat, Mahipal Singh;Park, Hye Yoon
    • Molecules and Cells
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    • v.42 no.4
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    • pp.356-362
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    • 2019
  • The binding of MS2 bacteriophage coat protein (MCP) to MS2 binding site (MBS) RNA stem-loop sequences has been widely used to label mRNA for live-cell imaging at single-molecule resolution. However, concerns have been raised recently from studies with budding yeast showing aberrant mRNA metabolism following the MS2-GFP labeling. To investigate the degradation pattern of MS2-GFP-labeled mRNA in mammalian cells and tissues, we used Northern blot analysis of ${\beta}$-actin mRNA extracted from the Actb-MBS knock-in and $MBS{\times}MCP$ hybrid mouse models. In the immortalized mouse embryonic cell lines and various organ tissues derived from the mouse models, we found no noticeable accumulation of decay products of ${\beta}$-actin mRNA compared with the wild-type mice. Our results suggest that accumulation of MBS RNA decay fragments does not always happen depending on the mRNA species and the model organisms used.

The Alcohol-inducible form of Cytochrome P450 (CYP 2E1): Role In Toxicology and Regulation of Expression

  • Novak, Raymond F.;Woodcroft, Kimberley J.
    • Archives of Pharmacal Research
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    • v.23 no.4
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    • pp.267-282
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    • 2000
  • Cytochrome P45O (CYP) 2E1 catalyzes the metabolism of a wide variety of therapeutic agents, procarcinogens, and low molecular weight solvents. CYP2E1-catalyzed metabolism may cause toxicity or DNA damage through the production of toxic metabolites, oxygen radicals, and lipid peroxidation. CYP2E1 also plays a role in the metabolism of endogenous compounds including fatty acids and ketone bodies. The regulation of CYP2E1 expression is complex, and involves transcriptional, post-transcriptional, translational, and post-translational mechanisms. CYP2E1 is transcriptionally activated in the first few hours after birth. Xenobiotic inducers elevate CYP2E1 protein levels through both increased translational efficiency and stabilization of the protein from degradation, which appears to occur primarily through ubiquitination and proteasomal degradation. CYP2E1 mRNA and protein levels are altered in response to pathophysiologic conditions by hormones including insulin, glucagon, growth hormone, and leptin, and growth factors including epidermal growth factor and hepatocyte growth factor, providing evidence that CYP2E1 expression is under tight homeostatic control.

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CYP1B1 Activates Wnt/β-Catenin Signaling through Suppression of Herc5-Mediated ISGylation for Protein Degradation on β-Catenin in HeLa Cells

  • Park, Young-Shin;Kwon, Yeo-Jung;Chun, Young-Jin
    • Toxicological Research
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    • v.33 no.3
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    • pp.211-218
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    • 2017
  • Cytochrome P450 1B1 (CYP1B1) acts as a hydroxylase for estrogen and activates potential carcinogens. Moreover, its expression in tumor tissues is much higher than that in normal tissues. Despite this association between CYP1B1 and cancer, the detailed molecular mechanism of CYP1B1 on cancer progression in HeLa cells remains unknown. Previous reports indicated that the mRNA expression level of Herc5, an E3 ligase for ISGylation, is promoted by CYP1B1 suppression using specific small interfering RNA, and that ISGylation may be involved in ubiquitination related to ${\beta}-catenin$ degradation. With this background, we investigated the relationships among CYP1B1, Herc5, and ${\beta}-catenin$. RT-PCR and western blot analyses showed that CYP1B1 overexpression induced and CYP1B1 inhibition reduced, respectively, the expression of $Wnt/{\beta}-catenin$ signaling target genes including ${\beta}-catenin$ and cyclin D1. Moreover, HeLa cells were treated with the CYP1B1 inducer $7,12-dimethylbenz[{\alpha}]anthracene$ (DMBA) or the CYP1B1 specific inhibitor, tetramethoxystilbene (TMS) and consequently DMBA increased and TMS decreased ${\beta}-catenin$ and cyclin D1 expression, respectively. To determine the correlation between CYP1B1 expression and ISGylation, the expression of ISG15, a ubiquitin-like protein, was detected following CYP1B1 regulation, which revealed that CYP1B1 may inhibit ISGylation through suppression of ISG15 expression. In addition, the mRNA and protein expression levels of Herc5 were strongly suppressed by CYP1B1. Finally, an immunoprecipitation assay revealed a direct physical interaction between Herc5 and ${\beta}-catenin$ in HeLa cells. In conclusion, these data suggest that CYP1B1 may activate $Wnt/{\beta}-catenin$ signaling through stabilization of ${\beta}-catenin$ protein from Herc5-mediated ISGylation for proteosomal degradation.

Experimental and Numerical Study of the Thermal Decomposition of an Epoxy-based Intumescent Coating (실험과 계산을 통한 에폭시 계열 내화도료의 열분해에 관한 연구)

  • Kim, Yangkyun
    • Fire Science and Engineering
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    • v.30 no.1
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    • pp.31-36
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    • 2016
  • This study investigates the characteristics of thermal decomposition of an epoxy-based intumescent paint using thermogravimetric analysis (TGA) and numerical simulation. A mathematical and numerical model is introduced to describe mass loss profiles of the epoxy-based intumescent coating induced by the thermal decomposition process. The decomposition scheme covers a range of complexity by employing simplified 4-step sequential reactions to describe the simultaneous thermal decomposition processes. The reaction rates are expressed by the Arrhenius law, and reaction parameters are optimized to fit the degradation behavior seen during thermogravimetric (TG) experiments. The experimental results show a major 2-step degradation under nitrogen and a 3-step degradation in an air environment. The experiment also shows that oxygen takes part in the stabilization of the intumescent coating between 200 and $500^{\circ}C$. The simulation results show that the proposed model effectively predicts the experimental mass loss as a function of time except for temperatures above $800^{\circ}C$, which were intentionally not included in the model. The maximum error in the simulation was less than 3%.

Assessment of organic matter biodegradation and physico-chemical parameters variation during co-composting of lignocellulosic wastes with Trametes trogii inoculation

  • Fersi, Mariem;Mbarki, Khadija;Gargouri, Kamel;Mechichi, Tahar;Hachicha, Ridha
    • Environmental Engineering Research
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    • v.24 no.4
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    • pp.670-679
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    • 2019
  • Lignin complexity molecule makes its biodegradation difficult during lignocellulosic wastes composting. So, the improvement of its biodegradation has usually been considered as an objective. This study aimed to determine the impact of Trametes trogii inoculation on organic matter and particularly on lignin and cellulose during green wastes co-composting with olive mill waste water sludge and coffee grounds. Three types of heaps (H1, H2 and H3) were investigated during 180 d. H3 and H2 were inoculated at the beginning of the process (t0) and 120 d later (t120), respectively while H1 was the control. Results showed the absence of pH stabilization in H3 during the first month. Also, in this period we observed a faster degradation of some easily available organic matter in H3 than in the other heaps. After 120 d, a better cellulose decomposition (25.28%) was noticed in H3 than in H1 and H2 (16%). Inoculation during the second fermentation phase induced supplementary lignin degradation in H2 with a percentage of 35% against 23 and 26% for H1 and H3, respectively. For all the runs, a Fourier Transform Infrared analysis showed aliphatic groups' decrease, OH groups' increase and lignin structural modification.

Improved Technologies to Produce Heterologous Proteins in Recombinant Escherichia coli. (재조합 대장균에서 외래단백질 발현을 위한 기술개발)

  • 박용철;권대혁;이대희;서진호
    • KSBB Journal
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    • v.16 no.1
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    • pp.1-10
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    • 2001
  • Escherichia coli has been used as an expression work horse for foreign genes. This article summarized recent development in genetic engineering techniques for overproduction of medical proteins and industrial enzymes. Special emphasis was placed upon research activities concerning folding and refolding of inclusion bodies at genetic and fermentation levels. Plasmid and mRNA stabilization, development of strong inducible promoters, modification of translational elements and reduction of rpoteolytic degradation were carried out to elevate an expression level of a target protein. Optimization of culture conditions, improvement of denaturation and renaturation steps and coexpression of molecular chaperones or foldase were accomplished to produce active proteins in soluble form. Fusion protein systems with selective separation and surface display technology were also performed in an effort to make the E. coli expression system more effective and versatile.

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Tribological Behavior of the Plasma Sprayed Fe$_2$O$_3$Added Zirconia Based Coatings ($Fe_2{O_3}$가 첨가된 지르코니아계 용사코팅층의 마모마찰 특성)

  • 신종한;임대순;안효석
    • Tribology and Lubricants
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    • v.16 no.2
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    • pp.84-90
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    • 2000
  • High Temperature wear behavior of plasma sprayed zirconia coatings containing up to 10 mol% of Fe$_2$O$_3$ were investigated. The wear test results showed that the addition of Fe$_2$O$_3$ particles to zirconia improved the wear resistance and lowered the coefficient of friction. Optimum concentration of Fe$_2$O$_3$ was about 5 mol%. Similar degradation behavior was observed at about 40$0^{\circ}C$ for both zirconia and Fe$_2$O$_3$ added zirconia coatings. The results indicated that stabilization of tetragonal phase and changes in mechanical properties such as hardness and toughness were responsible for tribological behavior of plasma sprayed zirconia contain Fe$_2$O$_3$.

A Study on Phenomena of Watertree and Dielectric Breakdown in XLPE (XLPE의 수트리와 절연파괴 현상에 관한 연구)

  • Lee, Sung-Il;Ryu, Sung-Lim;Park, Il-Kyu;Lee, Ho-Sik
    • Proceedings of the Korean Institute of Electrical and Electronic Material Engineers Conference
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    • 2001.11b
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    • pp.262-265
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    • 2001
  • In order to investigate the water tree degradation behavior on XLPE cable, direct voltage of 200 to 800V has been applied to the material at $50^{\circ}C\sim100^{\circ}C$. and the water tree property has been correlated with voltage and temperature in this study. The leakage current was shown to increase as temperature increased and the Ohm's law was generally satisfied in this experiment though some experimental errors were found. The leakage current was shown to decrease and reach to the stable state with time. It was also shown that the time for the stabilization of leakage current was lessened as voltage increased

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Aeration Control of Thermophilic Aerobic Digestion Using Fluorescence Monitoring

  • Kim, Young-Kee;Oh, Byung-Keun
    • Journal of Microbiology and Biotechnology
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    • v.19 no.1
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    • pp.93-98
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    • 2009
  • The thermophilic aerobic digestion (TAD) process is recognized as an effective method for rapid waste activated sludge (WAS) degradation and the deactivation of pathogenic microorganisms. Yet, high energy costs due to heating and aeration have limited the commercialization of economical TAD processes. Previous research on autothermal thermophilic aerobic digestion (ATAD) has already reduced the heating cost. However, only a few studies have focused on reducing the aeration cost. Therefore, this study applied a two-step aeration control strategy to a fill-and-draw mode semicontinuous TAD process. The NADH-dependent fluorescence was monitored throughout the TAD experiment, and the aeration rate shifted according to the fluorescence intensity. As a result, the simple two-step aeration control operation achieved a 20.3% reduction in the total aeration, while maintaining an effective and stable operation. It is also expected that more savings can be achieved with a further reduction of the lower aeration rate or multisegmentation of the aeration rate.