• Title/Summary/Keyword: ClpB

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Identification and Cloning of the ClpB Gene in Psychromonas arctica by Inverse PCR and Cassette PCR Technology

  • Choi, Ae-Ran;Na, Joo-Mi;Sung, Min-Sun;Im, Ha-Na;Lee, Kyung-Hee
    • Bulletin of the Korean Chemical Society
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    • v.31 no.4
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    • pp.887-890
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    • 2010
  • The family of ClpB protein is a molecular chaperone which protects cellular proteins from being aggregated upon exposure to severe environmental stresses in association with DnaK/DanJ/GrpE in the ATP-dependent manner. In a psychrophilic bacterium which survives at a subzero temperature, any functional role of cold-active ClpB protein can be rather crucial. In order to identify a ClpB encoding gene from a cold-adapted bacterium whose genome sequence has not been fully discovered, we have employed a series of PCR technologies, including a gradient PCR with homologous primers, an inverse PCR and a cassette PCR. The full sequence of PaclpB gene was successfully identified and compared with those of other psychrophilic species. We have further cloned the gene in E.coli expression systems and were able to induce PaClpB protein expression by IPTG, which help us understand a molecular mechanism for survival against extremely cold environments.

A study on the fading compensation scheme for wireless ATM systems (무선 ATM 시스템을 위한 페이딩 보상기법에 관한 연구)

  • Chung, Boo-Young;Kang, Young-Heung;Cho, Sung-Jun;Lee, Jin
    • The Journal of Korean Institute of Communications and Information Sciences
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    • v.22 no.12
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    • pp.2727-2735
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    • 1997
  • This paper describes a new fading compensation scheme for Wireless ATM(WATM) sytem using pilot symbols. We have carried out a simulation for bit error rate(BER) and cell loss probability(CLP) to analyze the performance of a new scheme. As the resutls, BER performance we improved by means of HEC only above 3 dB of $E_{b}/N_{0}$ in AWGN, but not at all in fading environment. However, by using a new fading compenstion scheme with HEC, BER performance degraded below to 1 dB of $E_{b}/N_{0}$ in AWGN, but BER and CLP performances in WATM system were improved remarkably, and meeted at CLP performance criterion for K=10 dB rician fading. Also, code rate will be high and harware size of codec will be small by using this new scheme with HEC OR FEC.

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Trichostatin A Protects Liver against Septic Injury through Inhibiting Toll-Like Receptor Signaling

  • Kim, So-Jin;Park, Jin-Sook;Lee, Do-Won;Lee, Sun-Mee
    • Biomolecules & Therapeutics
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    • v.24 no.4
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    • pp.387-394
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    • 2016
  • Sepsis, a serious clinical problem, is characterized by a systemic inflammatory response to infection and leads to organ failure. Toll-like receptor (TLR) signaling is intimately implicated in hyper-inflammatory responses and tissue injury during sepsis. Histone deacetylase (HDAC) inhibitors have been reported to exhibit anti-inflammatory properties. The aim of this study was to investigate the hepatoprotective mechanisms of trichostatin A (TSA), a HDAC inhibitor, associated with TLR signaling pathway during sepsis. The anti-inflammatory properties of TSA were assayed in lipopolysaccharide (LPS)-stimulated RAW264.7 cells. Polymicrobial sepsis was induced in mice by cecal ligation and puncture (CLP), a clinically relevant model of sepsis. The mice were intraperitoneally received TSA (1, 2 or 5 mg/kg) 30 min before CLP. The serum and liver samples were collected 6 and 24-h after CLP. TSA inhibited the increased production of tumor necrosis factor (TNF)-${\alpha}$ and interleukin (IL)-6 in LPS-stimulated RAW264.7 cells. TSA improved sepsis-induced mortality, attenuated liver injury and decreased serum TNF-${\alpha}$ and IL-6 levels. CLP increased the levels of TLR4, TLR2 and myeloid differentiation primary response protein 88 (MyD88) protein expression and association of MyD88 with TLR4 and TLR2, which were attenuated by TSA. CLP increased nuclear translocation of nuclear factor kappa B and decreased cytosolic inhibitor of kappa B ($I{\kappa}B$) protein expression, which were attenuated by TSA. Moreover, CLP decreased acetylation of $I{\kappa}B$ kinase (IKK) and increased association of IKK with $I{\kappa}B$ and TSA attenuated these alterations. Our findings suggest that TSA attenuates liver injury by inhibiting TLR-mediated inflammatory response during sepsis.

Mn$2^+$ dependent ClpL ATPase in Streptococcus pneumoniae

  • Park, Moo-Hyun;Kwon, Hyog-Young;Jung, Seung-Ha;Pyo, Suhk-Neung;Rhee, Dong-Kwon
    • Proceedings of the PSK Conference
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    • 2002.10a
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    • pp.336.1-336.1
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    • 2002
  • HSP100/Clp family functions as molecular chaperone and ATP dependent protease. The Streptococcus pneumoniae ClpL. a homologue of bacterial ClpB and yeast cytosolic HSP 104. is one of major heat shock proteins but its biochemical properties are unknown. In this study. ClpL in Streptococcus pneumoniaewas characterized using histidine tagged recombinant ClpL. When ATP hydrolysis activity was compared in the presence or absence of a variety of nucleotides or divalent ions. either ATP or Mn$2^+$ ion was found to increase significantly the rate of ATP hydrolysis. Furthermore. glutaraldehyde cross-linking and subsequent native-PAGE analfysis showed that ClpL forms dimer. but in the presence of 4 mM concentration of $Mn^{2+}$ion as a cofactor for ATP hydrolysis and oligormerization in vitro.

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Regulation of Activity of the Response Regulator RssB (Response Regulator RssB의 활성 조절)

  • Park, Hee Jeong;Bang, Iel Soo
    • Korean Journal of Microbiology
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    • v.49 no.3
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    • pp.215-220
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    • 2013
  • Against environmental stresses, many bacteria utilize the alternate sigma factor RpoS that induces transcription of the specific set of genes helpful in promoting bacterial survival. Intracellular levels of RpoS are determined mainly by its turnover through proteolysis of ClpXP protease. Delivery of RpoS to ClpXP strictly requires the adaptor protein RssB. The two-component-type response regulator RssB constantly interacts with RpoS, but diverse environmental changes inhibit this interaction through modification of RssB activity, which increases RpoS levels in bacteria. This review discusses and summarizes recent findings on regulatory factors in RssB-RpoS interactions, including IraD, IraM, IraP anti-adaptor proteins of RssB and phosphorylation of N-terminal receiver domain of RssB. New information shows that the coordinated regulation of RssB activity in controlling RpoS turnover confers efficient bacterial defense against stresses.

Mutation in clpxoo4158 Reduces Virulence and Resistance to Oxidative Stress in Xanthomonas oryzae pv. oryzae KACC10859

  • Cho, Jung-Hee;Jeong, Kyu-Sik;Han, Jong-Woo;Kim, Woo-Jae;Cha, Jae-Soon
    • The Plant Pathology Journal
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    • v.27 no.1
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    • pp.89-92
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    • 2011
  • Cyclic AMP receptor-like protein (Clp), is known to be a global transcriptional regulator for the expression of virulence factors in Xanthomonas campestris pv. campestris (Xcc). Sequence analysis showed that Xanthomonas oryzae pv. oryzae (Xoo) contains a gene that is strongly homologous to the Xcc clp. In order to determine the role of the Clp homolog in Xoo, a marker exchange mutant of $clp_{xoo4158}$ was generated. Virulence and virulence factors, such as the production of cellulase, xylanase, and extracellular polysaccharides (EPS) and swarming motility were significantly decreased in the $clp_{xoo4158}$ mutant. Moreover, the mutation caused the strain to be more sensitive to hydrogen peroxide and to over-produce siderophores. Complementation of the mutant restored the mutation-related phenotypes. Expression of $clp_{xoo4158}$, assessed by reverse-transcription realtime PCR and clp promoter activity, was significantly reduced in the rpfB, rpfF, rpfC, and rpfG mutants. These results suggest that the clp homolog, $clp_{xoo4158}$, is involved in the control of virulence and resistance against oxidative stress, and that expression of the gene is controlled by RpfC and RpfG through a diffusible signal factor (DSF) signal in Xanthomonas oryzae pv. oryzae KACC10859.

The Beneficial Effect of Trolox on Sepsis-Induced Hepatic Drug Metabolizing Dysfunction

  • Park, Sang-Won;Lee, Sun-Mee
    • Archives of Pharmacal Research
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    • v.27 no.2
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    • pp.232-238
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    • 2004
  • Trolox is a hydrophilic analogue of vitamin E. The aim of this study was to investigate its effects on hepatic injury, especially alteration in cytochrome P450 (CYP)-dependent drug metabolism during polymicrobial sepsis. Rats were subjected to polymicrobial sepsis by cecal ligation and puncture (CLP). The rats were treated intravenously with Trolox (2.5 mg/kg) or vehicle, immediately after CLP. Serum aminotransferases and lipid peroxidation levels were markedly increased 24 h after CLP. This increase was attenuated by Trolox. Total CYP content and NADPH-P450 reductase activity decreased significantly 24 h after CLP. This decrease in CYP content was attenuated by Trolox. At 24 h after CLP, there was a significant decrease in the activity of these CYP isozymes: CYP1A1, 1A2, 2B1, and 2E1. However, Trolox differentially inhibited the decrease in CYP isozyme activity. Trolox had little effect on the decrease in CYP1A1 activity but Trolox significantly attenuated decreases in CYP1A2 and 2E1 activities. In fact, Trolox restored CYP2B1 activity to the level of activity found in control rats. Our findings suggest that Trolox reduces hepatocellular damage as indicated by abnormalities in hepatic drug-metabolizing function during sepsis. Our data also indicates that this protection is, in part, caused by decreased lipid peroxidation.

Quality Characteristics and Antioxidative Effects of Cookie Prepareds with Curcuma longa L. Powder (울금 분말을 첨가한 쿠키의 품질특성과 항산화 효과)

  • Ju, Sung-Mee;Hong, Ki-Woon
    • Journal of the East Asian Society of Dietary Life
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    • v.21 no.4
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    • pp.535-544
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    • 2011
  • The purpose of this study was to investigate the quality characteristics and antioxidative effect of cookies prepared with Curcuma longa L. powder (0, 1, 2, 3, 4%) substituted for flour. The pH of cookie dough decreased significantly in response to the addition of all levels of Curcuma longa L. powder (CLP). However, there were no significant differences in dough values among the test groups. Furthermore, when the spread factor values were compared among the groups, they were found to be inversely proportional to the CLP concentration. In addition, Hunter's color L value significantly decreased with increasing CLP content, whereas the a and b values increased. According to textural measurements, substitution of 1~4% for CLP resulted in increased hardness, cohesiveness, springiness, and brittleness when the samples were compared to the control. The results of sensory evaluation showed that the cookies containing 2% CLP had the highest scores. The acid value, peroxide value, and thiobarbituric acid value were all lower in cookies with 2% and 3% CLP than in cookies made with 4% CLP and control cookies.

Quality Characteristics of Noodle (Garakguksu) with Curcuma longa L. Powder (울금가루를 첨가한 가락국수의 품질 특성)

  • Song, Seung-Heon;Jung, Hyun-Sook
    • Korean journal of food and cookery science
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    • v.25 no.2
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    • pp.199-205
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    • 2009
  • Wet noodles with different percentages of Curcuma Ionga L. powder(CLP) as an additive were generated and their cooking characteristics were evaluated. Wheat flour with 8% CLP had the highest water binding capacity and breakdown, while setback was reversed. Cooked noodle characteristics, weight, volume and tensile strength decreased as CLP content of wheat flour increased, but turbidity of the soup was reversed. L value of wet noodles was higher than cooked noodles, while b value, and texture of wet noodles were lower than cooked noodles. Hunter color value and texture measurements demonstrated decreasing L value, springiness and cohesiveness but increasing b value, hardness, gumminess and chewiness with increasing CLP content. In sensory evaluations, noodles made with 4% CLP were most highly preferred, while noodles made with 2% and 8% CLP were less preferred than CLP-free prepared noodles.

Effect of a-Tocopherol on Vasoregulatory Gene Expression during Polymicrobial Sepsis

  • Kim, Joo-Young;Park, Sang-Won;Lee, Sun-Mee
    • Proceedings of the Korean Society of Applied Pharmacology
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    • 2003.11a
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    • pp.77-77
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    • 2003
  • Sepsis remains common surgical problems with high morbidity and mortality despite improvement in the management for septic patient. Although hepatocellular dysfunction occurs during sepsis, the mechanism responsible for this remains unclear. In sepsis, a state of severe oxidative stress is encountered, with host endogenous antioxidant defenses overcome. Therefore, the aim of this study was to determine the effect of a-tocopherol (AT) vasoregulatory gene expression during polymicrobial sepsis. Rats were subjected to polymicrobial sepsis by cecal ligation and puncture (CLP). AT (15 mg/kg) was intraperitonealy injected for 3 days prior to CLP. Blood samples were taken 24 h after CLP for measurement of the extent of hepatocellular damage. Liver samples were taken for RT-PCR analysis of mRNA for genes of interest: endothelin-l (ET-l), its receptors $ET_{A}$ and $ET_{B}$, nitric oxide synthases (iNOS and eNOS), cyclooxygenase-2 (COX -2), heme oxygenase-l (HO-l), and tumor necrosis factor-$\alpha$ (TNF-$\alpha$). The activities of serum alanine aminotransferase and lipid peroxidation level were significantly increased; an increase which was prevented by AT pretreatment. CLP significantly increased the mRNA levels of ET-1 and $ET_{B}$; an increase that was prevented by AT pretreatment. However, the level of $ET_{A}$ mRNA significantly decreased after CLP; a decrease that was not prevented by AT pretreatment. There were significant increases in the mRNA expression of iNOS, HO-l and COX -2 in CLP groups. This increase was prevented by AT pretreatment. The expression of eNOS and TNF-$\alpha$ mRNA significantly increased in CLP, which was not prevented by AT pretreatment. Our findings suggest that there was an imbalanced vasoregulatory gene expression in sepsis, and AT ameliorates this change through its free radical scavenging activity.

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