• Title/Summary/Keyword: Clp

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Escherichia coli내의 ATP-dependent Clp효소의 ATPase 활성 연구 (Properties of ATPase Activity of ATP-dependent Clp Protease in Escherichia coli)

  • 김승호
    • 한국미생물·생명공학회지
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    • 제21권1호
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    • pp.30-35
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    • 1993
  • E.coli에서 발견된 ATP-dependent 효소인 Clp효소 중에서 Clp A의 ATPase 활성에 대한 영향을 검토하였다. Clp효소의 limiting amount으로 나타난 specific 활성은 일정하게 증가하는 효소의존성을 보였다. ATPase 활성을 나타내고 있는 ClP A는 casein에 의하여 활성화되어지며 2분자의 ATP가 결합하고 ATPase 활성을 나타내기 위한 ATP의 분해는 Clp효소의 단백질 분해 활성에 필요하다.

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Excherichia coli 에 존재하는 ATP -의존성 Clp 효소의 안정성 및 특성 (Stability and Characterization of the ATP-dependent Clp Protease from Escherichia coli )

  • 김승호
    • 미생물학회지
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    • 제30권6호
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    • pp.528-532
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    • 1992
  • 계면 활성제 특히 Triton X-100 의 첨가에 의해 ATP-의존성 효소의 subunit 인 Clp P 의 안정도를 높히고, 염의 첨가에 의한 효소 활성의 유지 정도와 Clp P 호소에 대한 생화학적 성질을 조사하였다. Clp A 가 가지고 있는 ATPase 활성을 나타내기 위하여서는 Clp P 의 단백질 분해 효소활성이 요구되며 Clp 효소의 다량체형성을 조사한 결과 ATP 가 존재하지 않을 경우에는 Clp A 와 Clp P 효소가 각각 용출하였으며 ATP 및 Mg 를 첨가하였을 때에는 complex 를 형성하여 분자량이 600,000 이상으로서 Clp P 효소는 Hexamer 로서 효소활성을 나타내며 Clp A 와 Clp P 의 다량체 형성이 요구되고 형성시에 ATP 가 관여하고 있는 것으로 나타났다.

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84-kDa의 폐렴구균 열충격단백질 ClpL의 Cloning 및 면역특성에 관한 연구 (Cloning and Immunological Characterization of the 84-kDa Heat Shock Protein, ClpL, in Streptococcus pneumoniae)

  • 권혁영;김용환;최혜진;박연진;표석능;이동권
    • Biomolecules & Therapeutics
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    • 제9권2호
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    • pp.79-87
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    • 2001
  • Heat shock proteins serve as chaperone by preventing the aggregation of denatured proteins and promote survival of pathogens in harsh environments. In this study, heat shock gene encoding a 84-kDa (p84) protein, which is one of the three major heat shock proteins in S. pneumoniae, was cloned and characterized. PCR with a forward primer derived from N-terminal amino acid sequence of the p84 and a reverse primer derived from the conserved second ATP-binding region of Clp family was used for amplification of the gene encoding the p84 and subsequently the PCR product was used for sequence determination. Sequence analysis of the p84 gene demonstrated that it is a member of ClpL. The deduced amino acid sequence of pneumococcal ClpL shows homology with other members of the Clp family, and particularly, even in variable leader region, with bovine Clp-like protein and L. lactis ClpL. S. pneumoniae clpL is the smallest clop member (701 amono acids) containing the two conserved ATP-binding regions, and hydrophilic N-terminal variable region of pneu-mococcal Clp ATPase is much shorter than any known Clp ATPases. Histidine tagged ClpL was overexpressed and purified from E. coli. Immunoblot analysis employing antisera raised against pneumococcus p84 demonstrated no cross-reactivity with Clp analog in Eschericha coli, Staphylococcus aureus and human HeLa cells. Preimmunization of mice with ClpL extended mice life partially but did not protect them from death.

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페렴구균 ClpP의 면역 교차 반응과 방어효과 (Cross-reactivity and Protective Immunity of Streptococcus pneumonieae ClpP)

  • 권혁영;이선숙;이순복;표석능;이동권
    • 약학회지
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    • 제48권1호
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    • pp.47-54
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    • 2004
  • ClpP is a stress-inducible protein and proteolytic subunit of the ATP-dependent Clp protease in prokaryotes and eukaryotes. Although its physiological roles in bacterial virulence were widely studied in various organsims, including Streptococcus pneumoniae, until now the immunological effect has not been investigated. Here, we have examined the cross reactivity of S. pneumoniae ClpP antibody with other organisms's cell lysate proteins. Although the protein sequence of S. pneumoniae ClpP was highly conserved among various organisms including human, the antibody rasised by S. pneumoniae ClpP was not cross-reacted with other organism's cell lysates, which were Saccharomyces cerevisiae , human lung A549 cell, Bacillus subtilis, Pseuomonas aeruginosa, E. coli, and Salmonella typhi. It was only reacted with S. pneumoniae and Lato-bacillus thermophilus. Thus we examined the immunoprotective effect of ClpP by immunizing mice with the purified ClpP. The mean survival time of mouse was significantly increased with the ClpP immunization. These results suggest that S. pneumoniae ClpP could be used as a vaccine candidate for prevention of S. pneumoniae infection.

Pseudomonas cepacia 균주가 생산하는 항진균성 Cyclic Lipopeptide의 생물학적 및 물리 화학적 특성 (Biological and Physico-chemical Properties of Antifungal Cyclic Lipopeptides Produced by Pseudomonas cepacia Strains)

  • 김성호;이민웅
    • 한국균학회지
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    • 제24권4호통권79호
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    • pp.310-321
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    • 1996
  • 토양으로부터 항진균 활성을 나타내는 5종류의 Pseudomonas cepacia 균주 AF027, AF069, AF2001, AF2011, SD02가 분리되었으며 이들은 각각 cyclic lipopeptide계 항진균 물질 CLP027A, CLP069A, Cepacidine A, CLP2011A, CLP02A를 생산하였다. 조사된 P cepacia 균주들은 배지중에 함유된 질소원 및 탄소원의 종류에 따라 항진균 물질 생산성에 현저한 차이가 있었는데, 질소원으로서는 polypeptone-S, 탄소원으로서는 glucose와 fructose가 생산성 증대에 가장 효과적이었다. 항진균물질 $CLP069A_1$$CLP069A_2$는 각각 분자량이 1215와 1199이며, 아미노산 조성, UV spectrum, IR spectrum 및 항진균 스펙트럼에 있어서, Xylocandin $A_1,\;A_2$와 일치하였으며, CLP027 $A_1$$A_2$ 또한 이러한 물리 화학적 특성이 Cepacidine A와 완전히 일치함으로써 동일 물질로 생각된다. $CLP2011A_1,\;A_2$$CLP02A_1,\;A_2$는 아미노산 조성 및 항진균 스펙트럼이 Xylocandin $A_1,\;A_2$와 다르고, Cepacidine $A_1,\;A_2$와는 항진균 스펙트럼에서 현저하게 차이가 났다. CLP2011A, CLP02A가 분자량, 아미노산 조성, UV spectrum, IR spectrum에서 Cepacidine A와 동일하지만 항진균 스펙트럼이 상이한 것은 이들이 다른 물질임을 시사하며, 이는 aliphatic side chain이 Cepacidine A의 그것과 다른 데에서 비롯된 분자구조의 상이함에서 기인된 것으로 추론된다.

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무선 ATM에서 CLP를 이용한 핸드오프 방식 (The Method of Handoff Using CLP on the WATM Network)

  • 김준배;황재문;강경식
    • 한국컴퓨터정보학회논문지
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    • 제6권3호
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    • pp.44-49
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    • 2001
  • 일반적으로 ATM/WATM(Asynchronous Transfer Mode/wireless ATM)에서는 서비스의 종류에 따라 추구하는 사용자의 서비스 품질은 다르며 실시간 비디오 데이터 서비스는 셀의 손실보다는 셀 전송 지연에 더 민감한 서비스이다. 이러한 서비스를 하는데 있어서 셀 손실 방지를 위주로 한 기존의 핸드오프 방식으로는 전송 지연의 측면에서 사용자의서비스 품질을 만족하는데 한계가 있었다. 본 논문에서는 실시간 VBR(Variable Bit Rate) 서비스에서 핸드오프 시 Forwarding해야 할 셀들 중 ATM 셀 헤더의 CLP(Cell Loss Priority)가 높은 셀은 폐기하고 낮은 셀만을 전송하는 방식을 제안하였다. 제안한 방식은 핸드오프 요구사항인 신속한 핸드오프에 부합하며, MPEG 프레임 중 중요하지 않은 프레임에 높은 CLP를 부여함으로써 QoS(Quality of Service)를 만족함을 시뮬레이션을 통하여 입증하였다.

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패혈증에서 Heat Shock Protein 70의 과도발현이 Inducible Nitric Oxide Synthase의 발현에 미치는 효과 (Effect of Heat Shock Protein 70 on Inducible Nitric Oxide Synthase during Sepsis in Rats)

  • 이용근;신효근;권운용;서길준;윤여규
    • Journal of Trauma and Injury
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    • 제21권1호
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    • pp.59-65
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    • 2008
  • Purpose: The aim of this study was to evaluate the effect of overexpression of heat shock protein 70 (HSP70) on the expression of inducible nitric oxide synthase and on the concentration of nitric oxide and to determine the mechanism for the relationship between HSP70 and inducible nitric oxide synthase (iNOS) in sepsis. Methods: Experiments were performed on male Sprague-Dawley rats, and sepsis was induced by using cecal ligation and puncture (CLP). Glutamine (GLN) or saline was administered 1 h after initiation of sepsis. We acquired serum and lung tissues from the rats 12 h or 24 h after initiation of sepsis. We analyzed the concentration of nitric oxide, the expression of HSP70 in the lung, and the gene expression of iNOS in the lung. Results: In CLP+GLN, glutamine given after initiation of sepsis enhanced the expression of HSP70 in the lung at 12 h (CLP+GLN vs. CLP:: $47.19{\pm}10.04$ vs. $33.22{\pm}8.28$, p = 0.025) and 24 h (CLP+GLN vs. CLP: $47.06{\pm}10.60$ vs. $31.90{\pm}4.83$, p = 0.004). In CLP+GLN, glutamine attenuated the expression of iNOS mRNA in the lung at 12 h (CLP+GLN vs. CLP: $4167.17{\pm}951.59$ vs. $5513.73{\pm}1051.60$, p = 0.025) and 24 h (CLP+GLN vs. CLP: $9,437.65{\pm}2,521.07$ vs. $18,740.27{\pm}8,241.20$, p = 0.016) and reduced the concentration of nitric oxide in serum at 12 h (CLP+GLN vs. CLP: $0.86{\pm}0.48$ vs. $3.82{\pm}2.53{\mu}mol/L$, p = 0.016) and 24 h (CLP+GLN vs. CLP: $0.39{\pm}0.25$ vs. $1.85{\pm}1.70{\mu}mol/L$, p = 0.025). Conclusion: The overexpression of HSP70 induced by the administration of glutamine in sepsis attenuated the gene expression of iNOS and reduced the concentration of nitric oxide.

물질안전보건자료 및 표시제도의 개선 방향 제안(2) - 유럽연합의 CLP 제도 고찰에 근거하여 (How to Improve the Reliability of MSDS and Labels?(2): A Review on the EU CLP(REGULATION(EC) No 1272/2008))

  • 김신범;이윤근;최영은
    • 한국산업보건학회지
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    • 제25권2호
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    • pp.115-125
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    • 2015
  • Objectives: The major objectives of this study are to review the EU CLP Regulations to propose ways of improving the reliability of MSDS and labels. Methods: To review the EU CLP Regulations, we used EU documents including directives and regulations on chemical management. In addition, we used EU governmental agency reports to illuminate the history and background of the CLP. We found the EU CLP's instruments for reliable hazard communication and evaluated the operations of the instruments. Results: EU CLP Regulations have four instruments for the EU CLP Regulations to make hazard communication reliable. These instruments are GHS, the harmonized CMR and respiratory sensitive substances classification list, C&L inventory and restriction of trade secrets. These are highly useful for achieving the objectives of REACH and CLP(no data-no market and changing the burden of proof). Conclusions: Changing the burden of proof is a key principle for achieving a society safe from hazardous chemicals. Chemical manufacturers and importers alone should bear the responsibility for reliable MSDS. We recommend benchmarking the EU CLP Regulations in order to change efficiently the burden of proof. Trade secrets should be limited to low-hazard substances and be approved by the government before the chemical product is on the market. Like the C&L inventory, chemical product information including substances identification and hazard properties should be notified, aggregated and be opened to public on the Internet. Finally, we recommend a MSDS registration system once again.

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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    • 제27권1호
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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.

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

  • Park, Moo-Hyun;Kwon, Hyog-Young;Jung, Seung-Ha;Pyo, Suhk-Neung;Rhee, Dong-Kwon
    • 대한약학회:학술대회논문집
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    • 대한약학회 2002년도 Proceedings of the Convention of the Pharmaceutical Society of Korea Vol.2
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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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