• Title/Summary/Keyword: glutathione transferase

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Combined transcriptome and proteome analyses reveal differences in the longissimus dorsi muscle between Kazakh cattle and Xinjiang brown cattle

  • Yan, XiangMin;Wang, Jia;Li, Hongbo;Gao, Liang;Geng, Juan;Ma, Zhen;Liu, Jianming;Zhang, Jinshan;Xie, Penggui;Chen, Lei
    • Animal Bioscience
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    • v.34 no.9
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    • pp.1439-1450
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    • 2021
  • Objective: With the rapid development of proteomics sequencing and RNA sequencing technology, multi-omics analysis has become a current research hotspot. Our previous study indicated that Xinjiang brown cattle have better meat quality than Kazakh cattle. In this study, Xinjiang brown cattle and Kazakh cattle were used as the research objects. Methods: Proteome sequencing and RNA sequencing technology were used to analyze the proteome and transcriptome of the longissimus dorsi muscle of the two breeds of adult steers (n = 3). Results: In this project, 22,677 transcripts and 1,874 proteins were identified through quantitative analysis of the transcriptome and proteome. By comparing the identified transcriptome and proteome, we found that 1,737 genes were identified at both the transcriptome and proteome levels. The results of the study revealed 12 differentially expressed genes and proteins: troponin I1, crystallin alpha B, cysteine, and glycine rich protein 3, phosphotriesterase-related, myosin-binding protein H, glutathione s-transferase mu 3, myosin light chain 3, nidogen 2, dihydropyrimidinase like 2, glutamate-oxaloacetic transaminase 1, receptor accessory protein 5, and aspartoacylase. We performed functional enrichment of these differentially expressed genes and proteins. The Kyoto encyclopedia of genes and genomes results showed that these differentially expressed genes and proteins are enriched in the fatty acid degradation and histidine metabolism signaling pathways. We performed parallel reaction monitoring (PRM) verification of the differentially expressed proteins, and the PRM results were consistent with the sequencing results. Conclusion: Our study provided and identified the differentially expressed genes and proteins. In addition, identifying functional genes and proteins with important breeding value will provide genetic resources and technical support for the breeding and industrialization of new genetically modified beef cattle breeds.

Effect of Ecklonia cava on the Blood Glucose, Lipids and Renal Oxidative Stress in Diabetic Rats (당뇨쥐에서 감태의 혈당, 혈청지질 개선효과 및 신장의 항산화효과)

  • Kim, Eun;Kim, Min-Sook;Kim, Se-Youn;Kim, Hyeon-A
    • Journal of the Korean Society of Food Culture
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    • v.23 no.6
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    • pp.812-819
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    • 2008
  • In this study, we assessed the effects of dietary supplementation with Ecklonia cava on blood glucose, lipid metabolism, and renal oxidative stress in streptozotocin (STZ)-induced diabetic rats. Male Sprague-Dawley rats were divided into a normal rat group fed on a control diet and diabetic rats fed on a control diet or supplemented with powder (15% w/w) or water extract of Ecklonia cava (2.5% w/w). Diabetes was induced by a single injection of STZ (60 mg/kg, ip) in citrate buffer. The animals were fed ad libitum with the experimental diet and water for 5 weeks. Dietary supplementation of Ecklonia cava powder and water extract was shown to reduce blood glucose levels in the diabetic rats, and the water extract was more effective than the powder. Dietary supplementation with Ecklonia cava also reduced LDL cholesterol and increased HDL-cholesterol levels in the diabetic rats. Renal glutathione S-transferase activity was increased in the diabetic rats as compared to the normal rats, but reverted to near control values as the result of dietary supplementation with Ecklonia cava. These results show that Eklonia cava exerts an anti-diabetic effect by improving blood glucose concentrations, LDL/HDL-cholesterol ratios, and antioxidative effects on the kidney in diabetic rats.

Protective effects of baicalein treatment against the development of nonalcoholic steatohepatitis in mice induced by a methionine choline-deficient diet

  • Jiwon Choi;Jayong Chung
    • Journal of Nutrition and Health
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    • v.56 no.6
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    • pp.589-601
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    • 2023
  • Purpose: Baicalein, a natural flavone found in herbs, exhibits diverse biological activities. Nonalcoholic steatohepatitis (NASH) is an irreversible condition often associated with a poor prognosis. This study aimed to evaluate the effects of baicalein on the development of NASH in mice. Methods: Male C57BL/6J mice were randomly divided into four groups. Three groups were fed a methionine-choline-deficient (MCD) diet to induce NASH and were simultaneously treated with baicalein (at doses of 50 and 100 mg/kg) or vehicle only (sodium carboxymethylcellulose) through oral gavage for 4 weeks. The control group was fed a methionine-choline-sufficient (MCS) diet without the administration of baicalein. Results: The baicalein treatment significantly reduced serum levels of alanine aminotransferase and aspartate aminotransferase, suggestive of reduced liver damage. Histological analysis revealed a marked decrease in nonalcoholic fatty liver activity scores induced by the MCD diet in the mice. Similarly, baicalein treatment at both doses significantly attenuated the degree of hepatic fibrosis, as examined by Sirius red staining, and hepatocellular death, as examined by the terminal deoxynucleotidyl transferase dUTP nick end labeling assay. Baicalein treatment attenuated MCD-diet-induced lipid peroxidation, as evidenced by lower levels of hepatic malondialdehyde and 4-hydroxynonenal, demonstrating a reduction in oxidative stress resulting from lipid peroxidation. Moreover, baicalein treatment suppressed hepatic protein levels of 12-lipoxygenase (12-Lox) induced by the MCD diet. In contrast, baicalein enhanced the activities of antioxidant enzymes such as superoxide dismutase, catalase, and glutathione peroxidase. Additionally, baicalein treatment significantly reduced hepatic non-heme iron concentrations and hepatic ferritin protein levels in mice fed an MCD diet. Conclusion: To summarize, baicalein treatment suppresses hepatic lipid peroxidation, 12-Lox expression, and iron accumulation, all of which are associated with the attenuation of NASH progression.

Wdpcp, a Protein that Regulates Planar Cell Polarity, Interacts with Multi‐PDZ Domain Protein 1 (MUPP1) through a PDZ Interaction (Planar cell polarity 조절단백질 Wdpcp와 multi-PDZ domain protein 1 (MUPP1)의 PDZ 결합)

  • Jang, Won Hee;Jeong, Young Joo;Choi, Sun Hee;Yea, Sung Su;Lee, Won Hee;Kim, Mooseong;Kim, Sang-Jin;Urm, Sang-Hwa;Moon, Il Soo;Seog, Dae-Hyun
    • Journal of Life Science
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    • v.26 no.3
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    • pp.282-288
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    • 2016
  • Protein-protein interactions regulate the subcellular localization and function of receptors, enzymes, and cytoskeletal proteins. Proteins containing the postsynaptic density-95/disks large/zonula occludens-1 (PDZ) domain have potential to act as scaffolding proteins and play a pivotal role in various processes, such as synaptic plasticity, neural guidance, and development, as well as in the pathophysiology of many diseases. Multi-PDZ domain protein 1 (MUPP1), which has 13 PDZ domains, has a scaffolding function in the clustering of surface receptors, organization of signaling complexes, and coordination of cytoskeletal dynamics. However, the cellular function of MUPP1 has not been fully elucidated. In the present study, a yeast two-hybrid system was used to identify proteins that interacted with the N-terminal PDZ domain of MUPP1. The results revealed an interaction between MUPP1 and Wdpcp (formerly known as Fritz). Wdpcp was identified as a planar cell polarity (PCP) effector, which is known to have a role in collective cell migration and cilia formation. Wdpcp bound to the PDZ1 domain but not to other PDZ domains of MUPP1. The C-terminal end of Wdpcp was essential for the interaction with MUPP1 in the yeast two-hybrid assay. This interaction was further confirmed in a glutathione S-transferase (GST) pull-down assay. When coexpressed in HEK-293T cells, Wdpcp was coimmunoprecipitated with MUPP1. In addition, MUPP1 colocalized with Wdpcp at the same subcellular region in cells. Collectively, these results suggest that the MUPP1-Wdpcp interaction could modulate actin cytoskeleton dynamics and polarized cell migration.

Comparison of Acute Toxicity of Pesticides between Carp(cyprinus carpio L.) and Israeli Carp(Cyprinus israeli carpio L.) (농약(農藥)에 의한 참잉어 및 이스라엘잉어의 급성독성비교(急性毒性比較))

  • Rim, Yo-Sup;Han, Seong-Soo
    • Korean Journal of Environmental Agriculture
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    • v.14 no.2
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    • pp.163-170
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    • 1995
  • This study was carried out to compare the acute toxicity(96hr) of 13 chemicals to carp (Cyprinus carpio L.) and israeli carp (Cyprinus israeli carpio L.) and the activities of acetylcholinesterase (AchE) and glutathione S-transferase (GST) in israeli carp exposed to five insecticides (diazinon, malathion, carbofuran, cartap, methomyl). $LC_{50}$ values of acute toxicity of the chemicals to israeli carp were endosulfan 0.0061ppm, captafol 0.041ppm, chlorothalonil 0.073ppm, butachlor 0.48ppm, captan 0.14ppm, carbofuran 1.13ppm, cartap 1.15ppm, diazinon 1.35ppm, nitrofen 3.72ppm, methomyl 4.39ppm, propanil 10.61ppm, malathion 11.78ppm and isoprothiolane 12.81ppm. The acute toxicity of endosulfan 0.0061ppm was 2100 times higher than that of isoprothiolane 12.81ppm. $LC_{50}$ values of acute toxicity of the chemicals to carp were endosulfan 0.0026ppm, captafol 0.062ppm, chlorothalonil 0.078ppm, captan 0.14ppm, and butachlor 0.47ppm, carbofuran 0.52ppm, nitrofen 0.58ppm, diazinon 0.81ppm, cartap 0.82ppm, methomyl 5.03ppm, propanil 10.67ppm, malathion 11.92ppm, and isoprothiolane 13.20ppm. The acute toxicity of endosulfan was 5,000 times higher than that of isoprothiolane. The toxicity of diazinon, carbofuran, cartap, endosulfan, and nitrofen to carp was approximately 2-6 times as high as that to israeli carp, but the toxicity of malathion, methomyl and captafol to israeli carp was slightly higher than that to carp. AchE activity was inhibited by 31% and 52% after 96hr’s exposure of israeli carp to diazinon and malathion respectively. GST activity in israeli carp was significantly induced by methomyl exposure for 96 hr.

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Cadms/SynCAMs/Necls/TSLCs Interact with Multi-PDZ Domain Protein MUPP1 (Cadms/SynCAMs/Necls/TSLCs와 multi-PDZ domain protein MUPP1 단백질의 결합)

  • Jang, Won Hee;Jeong, Young Joo;Choi, Sun Hee;Kim, Sang-Jin;Urm, Sang-Hwa;Moon, Il Soo;Seog, Dae-Hyun
    • Journal of Life Science
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    • v.24 no.12
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    • pp.1276-1283
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    • 2014
  • Cell adhesion molecules determine the cell-cell binding and the interactions between cells and extracellular signals. Cell-cell junctional complexes, which maintain the structural integrity of tissues, consist of more than 50 proteins including multi-PDZ domain protein 1 (MUPP1). MUPP1 contains 13 postsynaptic density-95/disks large/zonula occludens-1 (PDZ) domains and serves a scaffolding function for transmembrane proteins and cytoskeletal proteins or signaling proteins, but the mechanism how MUPP1 links and stabilizes the juxtamembrane proteins has not yet been elucidated. We used the yeast two-hybrid system to identify proteins that interact with PDZ domains of MUPP1. We found an interaction between MUPP1 and cell adhesion molecule 1 (Cadm1, also known as SynCAM1, Necl-2, or TSLC1). Cadm1 bound to the second PDZ domain of MUPP1. The carboxyl (C)-terminal end of Cadm1 has a type II PDZ-association motif (-Y-F-I) which was essential for the interaction with MUPP1 in the yeast two-hybrid assay. MUPP1 also bound to the C-terminal cytoplasmic tail region of other Cadm family members (Cadm2, Cadm3, and Cadm4). In addition, these protein-protein interactions were observed in the glutathione S-transferase (GST) pull-down assay and by co-immunoprecipitation. Anti-MUPP1 antibody co-immunoprecipitated Cadm1 and Cadm4 with MUPP1 from mouse brain extracts. These results suggest that MUPP1 could mediate interaction between Cadms and cytoskeletal proteins.

Ferritin, an Iron Storage Protein, Associates with Kinesin 1 through the Cargo-binding Region of Kinesin Heavy Chains (KHCs) (철 저장 단백질 ferritin과 kinesin 1 결합 규명)

  • Jang, Won Hee;Jeong, Young Joo;Lee, Won Hee;Kim, Mooseong;Kim, Sang-Jin;Urm, Sang-Hwa;Moon, Il Soo;Seog, Dae-Hyun
    • Journal of Life Science
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    • v.26 no.6
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    • pp.698-704
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    • 2016
  • The intracellular transport of organelles and protein complexes is mediated by kinesin superfamily proteins (KIFs). The first kinesin, kinesin 1, was identified as a molecular motor protein that moves various organelles and protein complexes along the microtubule rails in cells. Kinesin 1 is a tetramer of two heavy chains (KHCs, also called KIF5s) and two kinesin light chains (KLCs). KIF5s interact with many different proteins through their tail region, but their binding proteins have not yet been fully identified. To identify the interaction proteins for KIF5A, we performed yeast two-hybrid screening and found a specific interaction with ferritin heavy chain (Frt-h), which has a role in iron storage and detoxification. Frt-h bound to the amino acid residues between 800 and 940 of KIF5A and to other KIF5s in the yeast two-hybrid assay. The coiled-coil domain of Frt-h is essential for interaction with KIF5A. In addition, ferritin light chain (Frt-l) interacted with KIF5s in the yeast two-hybrid assay. In addition, these proteins showed specific interactions in the glutathione S-transferase (GST) pull-down assay. An antibody to KHC specifically co-immunoprecipitated Frt-h and Frt-l from mouse brain extracts. These results suggest the kinesin 1 motor protein may transport the ferritin complex in cells.

The GSTT1 Genotype as A Marker for Susceptibility to Lung Cancer in Korean Female Never-Smokers (한국인 비흡연 여성에서 폐암의 유전적 감수성 표지자로서의 GSTT1 유전자형)

  • Jang, Sang Soo;Jung, Chi Young;Lee, Sin Yeob;Lee, Jae Hee;Jeon, Hyo-Sung;Park, Sun Ha;Son, Ji-Woong;Lee, Eung Bae;Kim, Chang Ho;Kam, Sin;Park, Rang Woon;Kim, In-San;Jung, Tae Hoon;Park, Jae Yong
    • Tuberculosis and Respiratory Diseases
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    • v.54 no.5
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    • pp.485-494
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    • 2003
  • Background : Most previous studies regarding the role of GSTMl and GSTT1 on lung cancer risk have been focused mainly on male smokers. However, epidemiological characteristics, histologic types and risk factors are different in female and male lung cancers, we investigated the association between these genotypes and lung cancer risk in males and females separately. Materials and Methods : The study population consisted of 253 lung cancer (153 males and 100 females) and 243 controls (140 males and 103 females). GSTM1 and GSTT1 genotypes were determined by a multiplex PCR. Results : In the male population, neither GSTM1 nor GSTT1 null genotype showed significant difference between cases and controls. In the female population, the frequencies of GSTM1 null genotype showed no significant difference between cases and controls. However, the frequencies of GSTT1 null genotype was significantly higher in cases (70.3%) than controls (55.3%, odds ratio (OR)=2.18; 95% confidence interval (CI=l.21-3.93). When the female population was stratified by age and smoking status, the ORs for GSTT1 null genotype were significantly higher in subgroups of ${\leq}60$ years (OR=4.82; 95% CI=l.61-14.4) and never-smokers (OR=4.29; 95% CI=1.94-9.48) but not in subgroups of >60 years or smokers. When stratifying the female never-smokers by age, the ORs for GSTT1 null genotype were significantly higher in both age groups of ${\leq}60$ years (OR=7.64; 95% CI=2.00-29.2) and >60 years (OR=2.89; 95% CI=1.05-7.94). Conclusion : We found that GSTT1 null genotype was associated with an increased risk of lung cancer in Korean female never-smokers. This result suggests that GSTT1 null genotype could be used as a biomarker for genetic susceptibility to lung cancer in Korean female never-smokers.

CYP450 1A1 and p53 expression and DNA adduct formation in the liver of rats treated with a single dose of aflatoxins (아플라톡신을 간회 투여한 랫드의 간에서 CYP450 1A1, p53의 발현과 DNA adduct의 형성)

  • Lee, Beom Jun;Lee, Sook Jin;Kim, Tae Myoung;Kim, Dae Joong;Nam, Sang Yoon;Hyun, Sang Hwan;Kang, Jong Koo;Hong, Jin Tae;Kim, Cheul Kyu;Yun, Young Won
    • Korean Journal of Veterinary Research
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    • v.44 no.4
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    • pp.507-513
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    • 2004
  • Aflatoxins are produced mainly by Aspergillus flavus and Aspergillus parasiticus that grow in improperly stored cereals. Aflatoxin B1 ($AFB_1$) is a potent hepatocarcinogen in a variety of experimental animals including human beings. In spite of a high attention to the hepatocarcinogenecity of $AFB_1$, the relative toxicity of aflatoxins ($AFB_2$ and $AFG_1$) is not fully clarified. Sprague-Dawley male rats were orally administered with $AFB_1$, $AFB_2$, and $AFG_1$ at the dose of 250 ${\mu}g/kg$ (additionally including a dose of $1250{\mu}g/kg $ for $AFB_1$) body weight. Animals were then killed at 12, 24 or 48 hrs following aflatoxin exposure. Subsequently the immunohistochemical examination of p53, cytochrome p450 1A1 (CYP450 1A1), and glutathione-S-transferase placental form (GST-P) were performed. The level of the 8-OxodG in the liver was determined. Expressions of CYP450 1A1 and p53 were high in the liver of rats through 48 hrs after treatment of $AFB_1$ at the single dose of $250{\mu}g/kg $. This pattern was more clear as increasing doses. The treatment of $AFB_2$ and $AFG_1$ did not affect the expression of CYP450 1A1 but it caused weak expression of p53. The activity of GST were not found in the liver of rats treated with aflatoxins. The formation of 8-OxodG by $AFB_1$ increased in a dose-dependent manner up to 24 hrs after a single treatment of $AFB_1$ thereafter decreased to the level of control. The treatment of $AFB_2$ and $AFG_1$ did not affect the levels of 8-OxodG in the liver of rats with increasing time. These results in the present study indicate that $AFB_1$ among aflatoxins with low comparable levels is the most toxic as determined by early biomarkers such as CYP450 1A1, p53, GST-P, and 8-OxodG.

Inhibitory Effort of the N-terminal GST on the Tautomerase Activity of Macrophage Migration Inhibitory Factor (GST 융합 시스템에서 나타나는 macrophage migration inhibitory factor의 tautomerase 활성 저해에 관한 연구)

  • Kim Sang-Soo;Kim Kyung-Hee;Park Hyo-Jin;Hur Eun-hye;Rhim Hyangshuk
    • Journal of Life Science
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    • v.15 no.6 s.73
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    • pp.961-967
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    • 2005
  • Macrophage migration inhibitory fartor (MIF), known as a cytokine, is a multifunctional protein that is ubiquitously expressed in a variety of cells and tissues; however, enzymatic function of MIF still remains elusive in cells. In this study, we assessed details of the tautomerase activity of MIF. We established rapid purification condition for MIF by using pGEX system and compared the L-dopachrome tautomerase activity of GST-MIF, tMIF, and MIF. The results show that GST (glutathione S-transferase)-epitope tag or N-terminal amino acids flanking the essential $P^{2}$ almost completely abrogated L-dopachrome tautomerase activity of MIF. Subsequently, to determine whether the N-terminal tags have effects on oligomerization of MIF, protein cross-linking products were analyzed on $15\%$ SDS-PACE. The result demonstrates that N-terminal tags are dispensable for the formation of MIF's homooligomers. Thus, the results imply that exposure of If containing hydrophobic pocket in the active site is critical for L-dopachrome tautomerase activity of MIF. In addition, our study suggest that the MIF's tautomerase activity might be influenced by interacting with cellular partners.