• Microbiology and Biotechnology Letters
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• v.21 no.1
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• pp.30-35
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• 1993

Clp is a relatively abundant ATP-dependent protease found in E. coli. Its specific activity was proportional to the concentration of the limiting amount of Clp A and an excess amount of Clp P, and vice versa. Clp A has an intrinsic ATPase activity that is stimulated by casein, and contains a second site for binding A TP, in addition to the ATPase site. The modification of sulfhydryl groups in Clp A with reagents which have bulky groups such as N-phenylmaleimide led to nullifying both ATPase and protease activity. The same sites were modified by sulfhydryl reagents. It seems that the sulfhydryl groups of Clp A are not directly involved in catalysis. Since non-hydrolyzable analogs of ATP do not activate Clp, ATP hydrolysis may be essential for the proteolytic activity of Clp protease. Clp A and Clp P did not associate in the absence of nucleotide. The results suggest that the activity of the proteolytic component, Clp P, is regulated by the A TP-dependent cycling of Clp A between the activator form and the non-activator form.

• Korean Journal of Microbiology
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• v.30 no.6
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• pp.528-532
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• 1992

The ATP-dependent protease. Clp P from Esehaichia coli has been increase the stahility with or without detergent as Triton X-100 and NP-40 in the Clp P. The C]p P proteolytic activity was remained to 0.1 M salt by $Na^{-1}$, $K^{+}$, $Li^{+}$ but was inhihited by $SO_4^{2}$. An active ATPase site in Clp A is required for A TP-dependent proteolysis by Clp protease as

• Biomolecules & Therapeutics
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• v.9 no.2
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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.

• YAKHAK HOEJI
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• v.48 no.1
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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.

• The Korean Journal of Mycology
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• v.24 no.4 s.79
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• pp.310-321
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• 1996

Five strains AF027, AF069, AF2001, AF2011 and SD02 of Pseudomonas cepacia were isolated from soil, and the antifungal cyclic lipopeptides(CLP) i.e, CLP027A, CLP069A, Cepacidine A, CLP2011A and CLP02A were produced from each strains, respectively. Nitrogen and carbon sources in media were proved to be important factors for the production of CLP and among them, polypeptone-S, glucose and fructose were the most effective. It appeared that compounds CLP027A and CLP069A were identical with Cepacidine A and Xylocandine A, respectively. contain aspartic acid as amino acid component, are differentiated from Xylocandine A containing asparagine. Although molecular weight, amino acid composition and UV spectrum of CLP2011A and CLP02A are same with those of Cepacidine A, it is postulated that these compounds are not identical with Cepacidine A when the antifungal spectra and antifungal activity were compared to those of Cepacidine A.

• Journal of the Korea Society of Computer and Information
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• v.6 no.3
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• pp.44-49
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• 2001

In general. requirements for QoS are different according to the type of services in wireline and wireless ATM, and real-time video service is more sensitive to cell transmission delay than to cell loss. Existing handoff schemes emphasizing prevention of cell loss had limitations in cell transmission delay to satisfy QoS. In this paper, a novel scheme to transmit ATM cells with low CLP (when CLP = 0) prior to others and discarding cells with high CLP (when CLP = 1) in ATM cell header among cells to be forwarded during handoffs in real-time VBR service is proposed. The proposed scheme is proven to be suitable for the satisfaction of QoS and appropriate for fast handoffs by giving high CLP value to less meaningful MPEG frames through simulations.

• Journal of Trauma and Injury
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• v.21 no.1
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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.

• Journal of Korean Society of Occupational and Environmental Hygiene
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• v.25 no.2
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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.

• 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.

• 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.