• Title/Summary/Keyword: OGT

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Screening of Preventive Effects of Herbal Prescriptions in Rat Model of Arterial Thrombosis Induced by Ferric Chloride

  • Lee, Ki Mo;Bang, Jihye;Lee, In Sun;Heo, Eun Jung;Kim, Bu Yeo;Kang, Hyung Won;Lyu, Yeoung Su;Jeon, Won Kyung
    • The Journal of Korean Medicine
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    • v.34 no.2
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    • pp.20-28
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    • 2013
  • Objectives: This study examined the effects of 8 types of herbal prescriptions prescribed to alleviate dementia symptoms in a rat model of arterial thrombosis induced by ferric chloride ($FeCl_3$). Methods: Thirty minutes before 35% $FeCl_3$ treatment, SD rats were intraperitoneally injected with the 8 types of herbal prescriptions, respectively. We tested the effects of the herbal prescriptions on time to occlusion (TTO) in an arterial thrombosis model using a laser Doppler flow meter. In addition, thrombus weight (TW) and collagen fiber damages were evaluated in the same condition. Results: Herbal prescriptions showed the following rank-order based on their TTO: Chong-myung-tang (CMT) > modified Jangwonhan 02 (LMK02) > Toki-shakuyaku-san (TSS) ${\geq}$ Oren-gedoku-to (OGT) ${\geq}$ Yokukansan (YKS). In particular, CMT (100 mg/kg, i.p.) and LMK02 (100 mg/kg, i.p.) delayed the TTO the most ($14.83{\pm}0.98$ and $13.67{\pm}1.03$ min, respectively) compared with the vehicle group ($7.95{\pm}0.78$ min, P<0.001). In addition, CMT, LMK02, and OGT treatment ($0.63{\pm}0.01$, $0.66{\pm}0.02$ and $0.67{\pm}0.01$ mg/mm, respectively) significantly reduced thrombus weight compared with the vehicle treatment ($0.78{\pm}0.03$, P<0.001) and also alleviated collagen fiber damage (CMT; $28.40{\pm}2.22$%, LMK02; $30.79 {\pm} 4.07$%, OGT; $26.20{\pm}1.48$%) in the vessels injured by $FeCl_3$. Therefore, CMT and LMK02 showed the greatest preventive activity in rat model of arterial thrombosis induced by $FeCl_3$. Conclusions: These results provide experimental evidence for traditional use of herbal prescriptions, suggesting that CMT and LMK02 extracts could be used to prevent vascular injury and thrombosis in the early stages of dementia.

The Potential 'O-GlcNAc-P'om' ('O-GlcNAc-P'om'의 존재 가능성)

  • Moon, Il Soo;Lee, HyunSook;Lee, Hyung Jong
    • Journal of Life Science
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    • v.23 no.2
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    • pp.324-331
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    • 2013
  • The addition and removal of N-acetylglucosamine (GlcNAc) molecules on serine or threonine residues of a protein is called O-GlcNAcylation. This post-translational modification occurs on both cytoplasmic and nuclear protein, and is fast and reversible as comparable to phosphorylation. In contrast to the phospho-signaling cycles, this emerging moon-lightening signaling is cycled by only two enzymes, O-GlcNAc transferase (OGT) and O-GlcNAcase (OGA). The simple machinery is a good evolutionary adaptation of a cell for quick accommodation to continuously fluctuating intra- and extracellular microenvironments. Rather than "switching" on or off a specific proteins - this would be done by phosphorylation where numerous specific kinases and phosphatases are involved - O-GlcNAcylation would play a "rheostat" which would be much more delicately increase or decrease the efficacy of signal transductions in response to cellular nutrient and stress conditions. Interestingly, recent evidence indicates that O-GlcNAc is further modified by phosphorylation. The O-GlcNAc-P will upgrade the modulation efficiency of cellular processes to continuous 'analogue' level. So far, only one protein AP180 was reported to have O-GlcNAc-P on Thr310. But, proteomic data from our laboratory indicate that there are multiple O-GlcNAc-P proteins, constituting "O-GlcNAc-P'om". This will focus on the possibility of existence of "O-GlcNAc-P'om".

Glucosamine increases vascular contraction through activation of RhoA/Rho kinase pathway in isolated rat aorta

  • Kim, Do-Hyung;Seok, Young-Mi;Kim, In-Kyeom;Lee, In-Kyu;Jeong, Seong-Yun;Jeoung, Nam-Ho
    • BMB Reports
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    • v.44 no.6
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    • pp.415-420
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    • 2011
  • Diabetes is a well-known independent risk factor for vascular disease. However, its underlying mechanism remains unclear. It has been reported that increased influx of the hexosamine biosynthesis pathway (HBP) induces O-GlcNAcylation of proteins, leading to insulin resistance. In this study, we determined whether or not O-GlcNAc modification of proteins could increase vessel contraction. Using an endothelium-denuded aortic ring, we observed that glucosamine induced OGlcNAcylation of proteins and augmented vessel contraction stimulated by U46619, a thromboxane $A_2$ agonist, via augmentation of the phosphorylation of MLC20$MLC_{20}$, MYPT1(Thr855), and CPI17, but not phenylephrine. Pretreatment with OGT inhibitor significantly ameliorated glucosamine-induced vessel constriction. Glucosamine treatment also increased RhoA activity, which was also attenuated by OGT inhibitor. In conclusion, glucosamine, a product of glucose influx via the HBP in a diabetic state, increases vascular contraction, at least in part, through activation of the RhoA/Rho kinase pathway, which may be due to O-GlcNAcylation.

Correlation of saponarin content with biosynthesis-related gene expression in hulled and hulless barley (Hordeum vulgare L.) cultivars

  • Lee, HanGyeol;Park, Jae-Hyeok;Yoon, A Mi;Kim, Young-Cheon;Park, Chul Soo;Yang, Ji Yeong;Woo, So-Yeun;Seo, Woo Duck;Lee, Jeong Hwan
    • Journal of Plant Biotechnology
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    • v.48 no.1
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    • pp.12-17
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    • 2021
  • Saponarin found in young barley sprouts has a variety of beneficial biological and pharmacological properties, including antioxidant, hypoglycemic, antimicrobial, and hepatoprotective activities. Our previous work demonstrated that saponarin content was correlated with the expression levels of three biosynthetic pathway genes [chalcone synthase (HvCHS1), chalcone isomerase (HvCHI), and UDP-Glc:isovitexin 7-O-glucosyltransferase (HvOGT1)] in young barley seedlings under various abiotic stress conditions. In this study, we investigated the saponarin content and expression levels of three saponarin biosynthetic pathway genes in hulled and hulless domestic barley cultivars. In the early developmental stages, some hulled barley cultivars (Kunalbori1 and Heukdahyang) had much higher saponarin contents than did the hulless barley cultivars. An RNA expression analysis showed that in most barley cultivars, decreased saponarin content correlated with reduced expression of HvCHS1 and HvCHI, but not HvOGT1. Heat map analysis revealed both specific increases in HvCHS1 expression in certain hulled and hulless barley cultivars, as well as general changes that occurred during the different developmental stages of each barley cultivar. In summary, our results provide a molecular genetic basis for the metabolic engineering of barley plants to enhance their saponarin content.

Investigations on High Temperature Filtration System for Particle Removal using Ceramic Candle Filter in the Pilot Scale Vitrification Plant (유리화 공정에서의 세라믹 캔들 필터를 이용한 분진 제거목적의 고온 여과 시스템에 관한 연구)

  • 류보현;박승철;황태원;하종현
    • Proceedings of the Korea Air Pollution Research Association Conference
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    • 2003.05b
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    • pp.391-392
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    • 2003
  • 한전 원자력 환경기술원에서는 중ㆍ저준위 방사성폐기물 유리화 기술의 상용화 가능성을 입증하기 위한 유리화 실증설비를 건설하여 시험 중에 있으며 이 유리화 기술은 유도 가열식 저온로(Cold Crucible Melter, CCM)에 폐기물을 투입하는 기술로서 폐기물의 부피 축소 효과와 더불어 최종 고화물로 생성되는 폐기물의 침출율이 매우 낮은 장점을 지닌다. 이와 같은 유리화 공정은 기존의 소각처리에서와 같이 폐기물의 열적 산화과정에 의해 유해오염가스와 입자성 물질이 발생된다. 따라서 이를 처리하기 위해배기체 처리공정(Off Gas Treatment System, OGTS)을 설치하여 환경 배출기준(SO$_2$300ppm, NO$_2$ 200ppm, CO 600ppm, HCI 50ppm, 분진 100mg/Nm$^3$ 등)을 만족하도록 하였고 특히 입자성 물질은 후단 OGTS나 배관내 침적으로 인한 방사성 오염을 막기 위해 CCM 후단에서 효율적으로 제거되어야만 한다. (중략)

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Quercetin induces cell death in cervical cancer by reducing O-GlcNAcylation of adenosine monophosphate-activated protein kinase

  • Ali, Akhtar;Kim, Min Jun;Kim, Min Young;Lee, Han Ju;Roh, Gu Seob;Kim, Hyun Joon;Cho, Gyeong Jae;Choi, Wan Sung
    • Anatomy and Cell Biology
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    • v.51 no.4
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    • pp.274-283
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    • 2018
  • Hyper-O-GlcNAcylation is a general feature of cancer which contributes to various cancer phenotypes, including cell proliferation and cell growth. Quercetin, a naturally occurring dietary flavonoid, has been reported to reduce the proliferation and growth of cancer. Several reports of the anticancer effect of quercetin have been published, but there is no study regarding its effect on O-GlcNAcylation. The aim of this study was to investigate the anticancer effect of quercetin on HeLa cells and compare this with its effect on HaCaT cells. Cell viability and cell death were determined by MTT and terminal deoxynucleotidyl transferase-mediated deoxyuridine triphosphate nick-end labelling assays. O-GlcNAcylation of AMP-activated protein kinase (AMPK) was examined by succinylated wheat germ agglutinin pulldown and immunoprecipitation. Immunofluorescence staining was used to detect the immunoreactivitiy of O-linked N-acetylglucosamine transferase (OGT) and sterol regulatory element binding protein 1 (SREBP-1). Quercetin decreased cell proliferation and induced cell death, but its effect on HaCaT cells was lower than that on HeLa cells. O-GlcNAcylation level was higher in HeLa cells than in HaCaT cells. Quercetin decreased the expression of global O-GlcNAcylation and increased AMPK activation by reducing the O-GlcNAcylation of AMPK. AMPK activation due to reduced O-GlcNAcylation of AMPK was confirmed by treatment with 6-diazo-5-oxo-L-norleucine. Our results also demonstrated that quercetin regulated SREBP-1 and its transcriptional targets. Furthermore, immunofluorescence staining showed that quercetin treatment decreased the immunoreactivities of OGT and SREBP-1 in HeLa cells. Our findings demonstrate that quercetin exhibited its anticancer effect by decreasing the O-GlcNAcylation of AMPK. Further studies are needed to explore how quercetin regulates O-GlcNAcylation in cancer.

Culture Conditions of E. coli Harboring Human O-Linked N-Acetyl-${\beta}$-Glucosaminidase Gene and Enzymatic Properties (사람의 O-linked-N-acetyl-${\beta}$-D-glucosaminidase 유전자를 함유한 대장균의 배양조건과 효소학적 특성)

  • 강대욱;조용권;서현효
    • Korean Journal of Microbiology
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    • v.40 no.2
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    • pp.147-153
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    • 2004
  • Protein modification by N-acetyl-${\beta}$-D-glucosamine (O-G1cNAc) on the hydroxyl groups of Ser or Thr ubiq-uitously occurs in eukaryotic cells and is involved in many cellular phenomena. The level of O-G1cNAc-mod-ified protein is regulated by OGT and O-GlcNAcase enzymes. We have tried to produce recombinant O-GlcNAcase in E. coli as an effort to establish in vitro screening system for modulators of O-GlcNAcase. The culture conditions for improvement of O-GlcNAcase productivity, were as follows: induction temperature, $30^{\circ}C$; the concentration of L-arabinose, 0.02% and induction time, 5 hr. Under these culture conditions, E. coli cells containing O-GlcNAcase gene had no enzyme activity until up to 3 hr culture. However, O-GlcNAcase activity dramatically increased from 3 to 5 hr culture. It almost maintained the same level after 5 hr culture. Western blot analysis verified the amount of expressed O-GlcNAcase increased with culture time, being con-sistent with activity data. The optimal reaction condition determined in this study was as follows: protein quan-tity, $5{\mu}g$; reaction time, 30 min; reaction temperature, $45^{\circ}C$; substrate concentration, 2 mM; reaction pH, 6.5. Methanol had little effect on O-GlcNAcase activity and 90% of activity were retained at 10%. Only 15% resid-ual activity were detected at 5% of chloroform.

Understanding Comprehensive Transcriptional Response of Salmonella enterica spp. in Contact with Cabbage and Napa Cabbage

  • Lee, Hojun;Kim, Seul I;Park, Sojung;Nam, Eunwoo;Yoon, Hyunjin
    • Journal of Microbiology and Biotechnology
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    • v.28 no.11
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    • pp.1896-1907
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    • 2018
  • Salmonellosis is commonly associated with meat and poultry products, but an increasing number of Salmonella outbreaks have been attributed to contaminated vegetables and fruits. Enteric pathogens including Salmonella enterica spp. can colonize diverse produce and persist for a long time. Considering that fresh vegetables and fruits are usually consumed raw without heat treatments, Salmonella contamination may subsequently lead to serious human infections. In order to understand the underlying mechanism of Salmonella adaptation to produce, we investigated the transcriptomics of Salmonella in contact with green vegetables, namely cabbage and napa cabbage. Interestingly, Salmonella pathogenicity island (SPI)-1 genes, which are required for Salmonella invasion into host cells, were up-regulated upon contact with vegetables, suggesting that SPI-1 may be implicated in Salmonella colonization of plant tissues as well as animal tissues. Furthermore, Salmonella transcriptomic profiling revealed several genetic loci that showed significant changes in their expression in response to vegetables and were associated with bacterial adaptation to unfavorable niches, including STM14_0818 and STM14_0817 (speF/potE), STM14_0880 (nadA), STM14_1894 to STM14_1892 (fdnGHI), STM14_2006 (ogt), STM14_2269, and STM14_2513 to STM14_2523 (cbi operon). Here, we show that nadA was required for bacterial growth under nutrient-restricted conditions, while the other genes were required for bacterial invasion into host cells. The transcriptomes of Salmonella in contact with cabbage and napa cabbage provided insights into the comprehensive bacterial transcriptional response to produce and also suggested diverse virulence determinants relevant to Salmonella survival and adaptation.