• Title/Summary/Keyword: toxic proteins

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Toxic Effects of Catechol and 4-Chlorobenzoate Stresses on Bacterial Cells

  • Park, Sang-Ho;Ko, Yeon-Ja;Kim, Chi-Kyung
    • Journal of Microbiology
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    • v.39 no.3
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    • pp.206-212
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    • 2001
  • Catechol and 4-chlorobenzoate (4CBA) which are produced from the biodegradation of a variety of aromatic and chloroaromatics have been recognized as toxic to living organisms. In this study, the toxic effects of catechol and 4-chlorobenzoate on gram-positive and -negative bacteria were examined in terms of survival, morphology, change in fatty acids and membrane protein composition. The survival rate of the organisms during treatment for 6 h was decreased, as the concentration of each aromatic was increased. Escherichia coli and Pseudomonas cells treated with catechol and 4CBA at concentrations causing a significant decrease in their viability, showed destructive openings in their cell envelopes. Bacills subtilis treated with the aromatics were reduced in cell size and Staphylococcus aureus cells displayed irregular rod shapes with wrinkled surfaces. The bacterial cells treated with 20 mM catechol showed increases in unsaturated fatty acids, but several saturated fatty acids were decreased. In the E. coli cells treated with 20 mM catechol, inner membrane proteins of 150 kDa and 105 kDa were decreased. But several kinds of the inner and outer membrane proteins were increased. In B. subtilis treated with 20 mM catechol, several kinds of proteins were increased or decreased in membrane proteins.

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Analysis of Toxicity in Escherichia coli from the Expression of Human Purinergic Receptor $P2X_4$ (인간 퓨린수용체 $P2X_4$를 발현시킬 때 나타나는 대장균 독성의 원인분석)

  • Yu, Yon-Joo;Jung, Yun-A;Lim, Dong-Bin
    • Korean Journal of Microbiology
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    • v.47 no.1
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    • pp.7-13
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    • 2011
  • In general, expression of membrane protein in Escherichia coli is very toxic to the host organism, but the mechanism for the toxicity is not clear yet. Expression of human purinergic receptor $P2X_4$ was found to be extremely toxic to the host E. coli. We examined this toxicity by isolation and analysis of less toxic mutant proteins. We could isolate 30 less toxic mutants of $P2X_4$ after hydroxylamine mutagenesis. Western blot showed that all of them produced proteins smaller than the wild type $P2X_4$. DNA sequencing of two largest mutant proteins showed that they were lost its second transmembrane domain. Localization analysis of these mutant proteins showed that they are not in cytoplasmic membrane, but in inclusion bodies. These data showed that inactive truncated $P2X_4$ is not toxic to E. coli and membrane integration and functionality of $P2X_4$ may be needed to show host toxicity.

TOXIC EFFECTS OF 2,4-D AND OTHER AROMATIC COMPOUNDS ON BACTERIA, AND THEIR PROTECTIVE RESPONSES

  • Oh, Kye-Heon;Kim, Chi-Kyung
    • Proceedings of the Korean Society for Applied Microbiology Conference
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    • 2000.04a
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    • pp.116-123
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    • 2000
  • The purpose of this work was to investigate the induction of stress shock proteins (SSPs) in Burkholderia sp. YK-2 in response to 2,4-dichlorophenoxyacetic acid (2,4-D), and Pseudomonas sp. DJ-12 to benzoate, 4-chlorobenzoate (4-CBA), 4-hydroxybenzoate, and biphenyl. The SSPs, which contribute to the resistance of the cytotoxic effect of the toxic aromatic compounds including 2,4-D and 4-CBA, were induced at different concentrations of the compounds in exponentially growing cultures of Burkholderia sp. YK-2 or Pseudomonas sp. DJ-12. This response involved the induction of a 43 kDa DnaK and 41 kDa GroEL proteins in Burkholderia sp. YK-2, characterized by SDS-PAGE and Western blot using the anti-DnaK and anti-GroEL monoclonal antibodies. In Pseudomonas sp. DJ-12, 70 kDa DnaK and 60 kDa GroEL proteins was induced as SSPs, respectively. The total SSPs were analyzed by 2-D PAGE. Survival of Burkholderia sp. YK-2 or Pseudomonas sp. DJ-12 with time in the presence of different concentrations of the compounds was monitored, and viable counts paralleled the induction of the SSPs in these strains. Cells treated with the increased concentrations of toxic compounds showed some destructive openings on the cell envelopes.

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Purification of a Mosquitocidal Toxic Protein from B. thuringiensis strain H9B by Immuno-Affinity Chromatography (Immuno-Affinity Chromatography에 의한 B. thuringiensis H9B 균주의 모기살충성 내독소 단백질의 정제)

  • 김광현;배수장;이광배
    • Journal of environmental and Sanitary engineering
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    • v.12 no.2
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    • pp.59-64
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    • 1997
  • For purification of a 70kDa toxic protein of mosquitocidal delta-endotoxin from B. thuringiensis strain H9B, immuno-affinity chromatography was performed. After separation of 70kDa toxic proteins from the delta-endotoxin of the strain H9B on SDS-PAGE, the 70kDa toxic protein was subcutaneously injected into rabbit for making a polyclonal antibody. A anti-70kDa toxic protein was purified by a column chromatography packed with protein A-sepharose 4B gels. The 70kDa toxic protein from delta-endotoxin of the strain H9B was also purified by an immuno-affinity chromatography packed with CNBr-activated sepharose 4B gels conjugated anti-70kDa toxic protein after elution with 1/10M citric acid-1/5M Na$_{2}$HPO$_{4}$ buffer(pH3.2) containing 0.5M NaCl. The 70kDa toxic protein was purified through only one step-separation system, was demonstrated by SDS-PAGE and immunoblot.

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PEGYLATION: Novel Technology to Enhance Therapeutic Efficacy of Proteins and Peptides (PEG 접합: 단백질 및 펩타이드 치료제의 약효를 증가시키는 새로운 기술)

  • Park, Myung-Ok;Lee, Kang-Choon
    • Journal of Pharmaceutical Investigation
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    • v.30 no.2
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    • pp.73-83
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    • 2000
  • Polyethylene glycol (PEG) is a water soluble, biocompatible, non-toxic polymer and PEGylation is a well established technique for the modification of therapeutic proteins and peptides. PEG-protein drugs have been extensively studies in relation to therapies for various diseases: cancer, inflammation and others. The covalent attachment of PEG to proteins and peptides prolonged plasma half-life, reduced antigenicity and immunogenicity, increased thermal and mechanical stability, and prevented degradation by enzymes. Several chemical groups for general and site specific conjugation have been exploited to activate PEG for amino group, carboxyl group, and cysteine groups. PEGylation of many proteins and peptides have been studied to enhance their properties for the potential uses. Also, the different positional isomers in several PEG-proteins have shown the difference in vivo stability and biological indicating that the site of PEG molecule attachment is one of the important factor to develop PEG-proteins as potential therapeutic agents.

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Comparative Study of Processed (Shodhit) and Unprocessed Seeds of 'Gunja'-Abrus precatorius L.

  • Gautam, D.N. Singh;Singh, P.N.;Mehrotra, Shanta
    • Natural Product Sciences
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    • v.5 no.3
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    • pp.127-133
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    • 1999
  • 'Gunja' is attributable to the plant Abrus precatorius L. (Leguminosae). Three forms with red, brown and another with white seeds are known to occur in this species and are employed for different therapeutic uses viz. as purgative, emetic, aphrodisiac, tonic and also as an abortifacient. According to some Ayurvedic literature the seeds are poisonous and should be given to the patients after proper processing ('Shodhan'). A comparative study of various phytochemical parameters, namely, percentage of successive extractives, total proteins, tannins, total ash and acid insoluble ash of these three forms of the processed (with cow's milk and Kanji) and unprocessed seeds was done. TLC and densitometric scanning of successive extractives was also carried out to serve as markers for processed and unprocessed seeds. The percentage of proteins, tannins, alcohol and water soluble extractives decreased in the processed material. Besides, their acute toxicity, CNS activity were also studied in albino mice and it was found that white seeds are more toxic as compared to the red and brown. The toxic effect was reduced with the processing. Further, the 'Kanji' processed seeds are less toxic than the milk processed one.

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몇 가지 PBTs (Persistent, Bioaccumulative, Toxic Chemicals)가 생태계 곤충에 미치는 영향

  • Lee Seun Yeong;Kim Yong Gyun
    • Proceedings of the Korea Society of Environmental Biology Conference
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    • 2002.11a
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    • pp.123-126
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    • 2002
  • Pollutants that are persistent, bioaccurnulative, and toxic have been linked to numerous adverse effects in human and animals, PBTs include heavy metals, polychlorinated biphenyls (PCBs), dioxins, polycyclic aromatic compounds (PACs) in addition to pesticides. This study focuses on toxic effects of the PBTs except pesticides on insects. Eight PBTs were selected from subgroups: three heavy metals (Pb, Hg, and Cd), two PCB mixtures (Aroclor mixtures 1 and 2), 2,3,7,8-tetrachlorodibenzo-p-dioxin, two monophenols (4-octylphenol and 4-nonylphenol), and tetrabutyltin, Beet armyworm, Spodoptera exigua, was used as test target insect species. Three physiological markers (metamorphosis, immune reaction, and follicle patency) were assessed in each exposure to different doses of the PCBs. Heat-shock proteins as molecular markers were also analyzed in response to the PCBs. All tested PBTs were toxic to metamorphosis from larvae to pupae when they were applied with diet. Two PCB mixtures were the most toxic compounds in this assay by giving significant toxicity at 0.005 ppm, while others had from 10 to 1000 ppm. Dioxin (0.1 ppb), tetrabutyltin (0.1 ppb), Pb (10 ppb), and Hg (0,01 ppb) were potent to inhibit immune reactions analyzed by inducing phenoloxidase activity and blocked phospholipase $A_2$ enzyme, Tetrabutyltin and dioxin significantly induced follicle cell patency, but their effects were lower than that of endogenous juvenile hormone, Dioxin, Pb, Hg, and Cd could induce the expression of heat shock proteins that were detected by immunoblotting against human HSP70 monoclonal antibody. HSP78 and HSP80 were upregulated in response to the PBTs. This expression was detected from the fat body and epidermis at as fast as 4h after injection. All these results clearly suggest that PBTs give significant ecotoxicity to insects that are valuable organisms in our environment.

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Therapeutic implication of autophagy in neurodegenerative diseases

  • Rahman, Md. Ataur;Rhim, Hyewhon
    • BMB Reports
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    • v.50 no.7
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    • pp.345-354
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    • 2017
  • Autophagy, a catabolic process necessary for the maintenance of intracellular homeostasis, has recently been the focus of numerous human diseases and conditions, such as aging, cancer, development, immunity, longevity, and neurodegeneration. However, the continued presence of autophagy is essential for cell survival and dysfunctional autophagy is thought to speed up the progression of neurodegeneration. The actual molecular mechanism behind the progression of dysfunctional autophagy is not yet fully understood. Emerging evidence suggests that basal autophagy is necessary for the removal of misfolded, aggregated proteins and damaged cellular organelles through lysosomal mediated degradation. Physiologically, neurodegenerative disorders are related to the accumulation of amyloid ${\beta}$ peptide and ${\alpha}-synuclein$ protein aggregation, as seen in patients with Alzheimer's disease and Parkinson's disease, respectively. Even though autophagy could impact several facets of human biology and disease, it generally functions as a clearance for toxic proteins in the brain, which contributes novel insight into the pathophysiological understanding of neurodegenerative disorders. In particular, several studies demonstrate that natural compounds or small molecule autophagy enhancer stimuli are essential in the clearance of amyloid ${\beta}$ and ${\alpha}-synuclein$ deposits. Therefore, this review briefly deliberates on the recent implications of autophagy in neurodegenerative disorder control, and emphasizes the opportunities and potential therapeutic application of applied autophagy.

Degradation of Collagens, Immunoglobulins, and Other Serum Proteins by Protease of Salmonella schottmulleri and its Toxicity to Cultured Cells

  • Na, Byoung-Kuk;Kim, Moon-Bo;Song, Chul-Yong
    • Journal of Microbiology
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    • v.34 no.1
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    • pp.95-100
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    • 1996
  • The effect of the extracellular protease of Salmonella schottmulleri on human serum constituents such as immunoglobulins, hemoglobin and lysozyme and tissue constituents such as fibronectin and collagens was investigated. This protease degraded collagens (type I and III), fibronectin and serum proteins such as human hemoglobin and lysozyme. Bovine serum albumin was degraded slightly. Thus, the present study suggested the possibility that this protease is not only played an important role in invasion of S. schottmulleri by degrading the constituent proteins such as collagens and fibronectin but also induced complications observed in septicemia and chronic infections by degrading the serum proteins. This protease is also capable of degrading defence-oriented humoral proteins, immunoglobulins (IgG and IgM). Furthermore, it is toxic to HEp-2 cells. These findings clarified the possible role of Salmonella protease as a virulence factor in the pathogenesis of Salmonella infections.

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Determination of Plasmids Encoding Crystal Toxic Protein Gene in Bacillus thuringiensis var kurstaki HD-1 (Bacillus thuringiensis var kurstaki HD-1의 내독소 단백질 유전에 관여하는 plasmid의 결정)

  • 김철영;김상현
    • Journal of Sericultural and Entomological Science
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    • v.35 no.2
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    • pp.120-128
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    • 1993
  • The objective of this study is to identify plasmids of Bacillus thuringiensis var. kurstaki HD-1(B. t k HD-1) toxic to lepidopteran larvae. The results from agarose gel electrophoresis indicated that the bacterium contained 9 plasmids with approximate sizes of 1.4, 4.9, 5.4, 9.3, 10, 29, 44, 52, and 150 megadaltons(Md). By treating the wild type of B. t k HD-1 with either SDS or EtBr as curing agent, 26 cured mutants of the bacterium were obtained, 9 of them were crystallifereous(cry+) and the others acrystallifereous(cry-). Plasmids from B. t k HD-1 were transferred to B. cereus 569 strR cry- recipients(Bc569 M1). Among 13 isolates of Bc569 M1 transcipient, 11 of them were capable of producing the crystal toxic proteins. The plasmid patterns of Bc569 M1 transcipients and partially curved mutants of B. t k HD-1 on agarose gel electrophoresis suggested that the 29 and 44Md plasmids should be involved in the production of crystalline toxic proteins.

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