• 제목/요약/키워드: ClpL

검색결과 25건 처리시간 0.026초

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

  • 주성미;홍기운
    • 동아시아식생활학회지
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    • 제21권4호
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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.

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.

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

  • 송승헌;정현숙
    • 한국식품조리과학회지
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    • 제25권2호
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    • pp.199-205
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    • 2009
  • 울금가루를 새로운 식품소재로 활용하고자 하는 연구의 일환으로 울금가루 첨가량을 달리하여 국수를 제조하고 그 품질특성을 조사하였다. 밀가루와 울금가루 혼합분의 물결합력과 breakdown은 울금가루 8% 첨가구에서 가장 높았으나, setback은 울금가루 8% 첨가구에서 가장 낮았다. 울금가루 첨가 국수의 조리 특성에서 무게, 부피 및 인장강도는 울금가루 8% 첨가구에서 가장 낮았고, 대조구에서 가장 높았으나, 탁도는 울금가루 8% 첨가구에서 가장 높았으며, 울금가루 첨가량이 증가할수록 높았다. 국수의 조리 전과 조리 후의 색도 및 물성의 변화에서 L값은 조리 전 생국수가 조리된 국수보다 높았으나, 색도의 a값 및 b값과 물성에서는 조리 전 생국수보다 조리된 국수에서 높았다. 색도의 L값과 물성의 탄력성 및응집성은 울금가루 첨가량이 증가할수록 낮아졌고, 색도의 a값 및 b값과 물성의 경도, 점착성 및 씹힘성은 울금가루 첨가량이 증가할수록 높아졌다. 울금가루 첨가량에 따른 관능검사에서 울금가루 4% 첨가구의 기호도가 가장 높았고, 울금가루 2% 및 8% 첨가구는 대조구보다 낮은 기호도를 보였다.

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

  • Kim, Joo-Young;Park, Sang-Won;Lee, Sun-Mee
    • 한국응용약물학회:학술대회논문집
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    • 한국응용약물학회 2003년도 Annual Meeting of KSAP : International Symposium on Pharmaceutical and Biomedical Sciences on Obesity
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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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CLP, Dhn5 유전자의 도입에 의한 고비사막 자생식물 Artemisia adamsii의 내건성 및 내동성 증진 (Transformation of Artemisia adamsii, Endemic to a Gobi Desert, with CLP, Dhn5 to Enhance Environmental Stress Tolerance)

  • 한규현;황철호
    • Journal of Plant Biotechnology
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    • 제30권4호
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    • pp.315-321
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    • 2003
  • 고비사막 접경지역의 식물인 Artermisia adamsii의 내건성 및 내동성 증진을 위해 조직배양과 CLP및 Dhn5유전자의 형질전환을 수행하였다. 다양한 호르몬 농도 조건에서의 성장을 확인한 결과, 0.05mg/L의 NAA와 0.5mg/L의 BAP조건과 0.1mg/L의 NAA와 0.5mg/L의 BAP가 포함된 배지의 암조건 하에서 최적의 캘러스 생장을 확인하였으며 유전자 도입 및 유전자의 발현이 확인된 캘러스가 세포 수준에서도 내건성 및 내동성이 증진되었음을 확인하였다. 그러나 Atremisia속 다른 식물과 다르게 조직절편에서 직접 기관분화를 유도하거나 캘러스를 통한 식물체 재생에 어려움이 있어 식물체 수준에서 형질전환에 따른 환경스트레스 내성의 증진을 확인하지 못하고 있다. 앞으로 진행될 A. adamsii의 식물체 재생에 대한 연구를 통해 내동성 및 내건성이 증진된 식물체를 육성하여 고비사막 지역적응력을 조사할 예정이다.

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.

감귤류 과피 추출물의 항염증 효과 (Anti-Inflammatory Effect of Peel Extracts from Citrus Fruits)

  • 이숙현;서석종;이경혜;양종범;최성업;박성수
    • 한국식품위생안전성학회지
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    • 제28권4호
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    • pp.342-348
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    • 2013
  • 본 연구에서는 감귤류 중 온주밀감, 레몬, 한라봉의 과피추출물(PE)에 대한 항염증 효과를 조사하였다. 감귤류 세품종 과피추출물(PE)에 대한 murine macrophage cell line Raw 264.7세포를 배양하여 세포독성, NO생성량, 세포내 염증관련 단백질 발현 양상을 분석하였다. 그 결과 CUP, CHP, CLP 처리구의 경우 2 mg/mL 이하의 농도에서는 세포생육을 저해하지 않았다. LPS (100 ng/mL)를 처리한 세포의 NO생성량을 LPS를 처리하지 않은 대조구와 비교했을 때 상당량이 증가했음을 알 수 있었고, 추출물을 농도별로 처리했을 때 농도가 증가할수록 NO생성량은 감소하였다. 각 처리구에서 각 1 mg/mL 농도에서는 CUP > CHP > CLP 순으로 NO 저해활성이 강하게 나타났다. Western blot의 결과상으로는 CUP와 CHP의 iNOS 단백질 저해활성이 비슷한 강도로 나타남을 확인하였다. COX-2 효소의 저해활성은 CUP > CHP > CLP의 순으로 나타났다. 따라서 CUP, CHP, CLP 처리구는 모두 항염증 작용을 가지고 있으며, 또한 CUP 처리구가 세 개의 추출물 중에서 가장 저해활성이 강하였다.