• Title/Summary/Keyword: shock proteins

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Expression of Heat Shock Protein Protein 70 in Umbilical Vein Endothelial Cells Infected by Staphylococcus aureus

  • Chang, Hyun-Ah;Chang, Jun-Keun;Kim, Jong-Won;Kim, Mal-Nam
    • Journal of Microbiology and Biotechnology
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    • v.10 no.2
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    • pp.137-142
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    • 2000
  • Environmental stres is known to induce heat shock proteins (HSPs) in eukaryotic cells. However, the induction of HSPs in host cells by microbial infection has not yet been well explained. Staphylococcus aureus (S. aureus) is one of the major pathogens in the pathogenesis of endovascular diseases such as infective endocarditis. In this study, the synthesis of stress-inducible 70 kDa HSP was investigated in the endothelial cells (ECs) after 3 h to 20 h of incubation with S. aureus. The dffect of S. aureus infection on the expression of HSP70 in cultured ECs was analyzed using laser scanning confocal microscopy (LSCM). The increase of HSP70 expression in ECs infected by S. aureus ($10^4{\;}cfu/ml$) for 20 h was 1.1-fold higher than that in heat shock treated ECs and 2.2-fold higher than that in untreated cells. Heat shock is known to induce intranucleus HSP70 expression in mammalian cells, whereas the S. aureus infection induced perinuclear expression in ECs as observed by LSCM. Consequently, the expression of HSP70 in ECs plays an important role in the pathogenesis of endovascular infection.

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Analysis of Antifungal Proteins in Streptomyces sp. SAR01 (Streptomyces sp. SAR01 균주에서의 항진균 관련 단백질 분석)

  • 이영근;김재성;조규성;장병일;추철형
    • Korean Journal of Environmental Biology
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    • v.20 no.3
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    • pp.237-244
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    • 2002
  • To analyze proteins related to antifungal activity, SAR01 strain was isolated from seaweed and identified as Streptomyces sp. from the result of FAME (fatty acid methyl ester) analysis. The isolated strain had antifungal activities against T species of plant pathogenic fungi. Antifungal activity deficient mutant (SAR 535) of Streptomyces sp. SAR01 was induced by gamma radiation $(^{60}Co,\;5kGy)$. By 2 D electrophoresis analysis, 6 protein spots were found in wild strain (SAR01) but these spots disappeared in mutant strain (SAR535). Among them, 5 proteins showed similarities to heat shock protein 70(HSP70), Fe-containing superoxide dismutase II (Fe- SODII), ribosome recycling factor (RRF), 10 kDa chnperonin (GroES) and inorganic pyrophosphatase (PPAse), respectively. It suggested that the above 6 proteins could be closely related to the antifungal activity of Streptomyces sp. SAR01.

Molecular chaperone as a sophisticated intracellular membership (세포내인자로서의 정교한 기능을 하는 molecular chaperone)

  • 권오유;송민호
    • Journal of Life Science
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    • v.8 no.2
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    • pp.226-226
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    • 1998
  • Discovery of molecular chaperone has stimulate cell biologists and thus made it possible to re-examine the processes whereby proteins achieve and maintain their functional conformations within living cells. the term ‘Molecular chaperone’ was first coined to describe one particular protein involved in the assembly of nucleosomes, but the term has now been extended to describe the function of a wide variety of proteins that assist protein transport across membranes, folding of nascent polypeptide, the assembly and disassembly of oligomeric structures, and the recovery or removal of proteins damaged by various environmental stresses including heat shock. Progress of molecular chaperone research is still limited by the lack of 3-dimensional structural information and detailed interacts with taget proteins in the cell. However, several laboratories around the world are attempting to extend our knowledge on the functions of molecular chaperone, and such efforts seem justified to finally provide the answers to the most burning questions shortly.

Evaluation of Thermo Tolerance of 'Nistari' an Indigenous Strain of Multivoltine Silkworm, Bombyx mori L.

  • Moorthy, S.M.;Das, S.K.;Mukhopadhyay, S.K.;Mandal, K.;Urs, S. Raje
    • International Journal of Industrial Entomology and Biomaterials
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    • v.15 no.1
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    • pp.17-21
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    • 2007
  • An indigenous multivoltine silkworm, Nistari was evaluated for their thermo tolerance by exposing the larvae to various temperature regimes for eight hours. Among different temperature exposed, this strain has significant tolerance at $32^{\circ}C$. Analysis of heat shock protein revealed the expression of 70 kDa and 64 kDa polypeptides in fat body and midgut tissues. Interestingly esterase isozyme pattern in midgut showed characteristic expression of Est-1 and Est-3 at different temperatures signifying role in heat and cold shock.

Analysis of heat, cold or salinity stress-inducible genes in the Pacific abalone, Haliotis discus hannai, by suppression subtractive hybridization

  • Nam, Bo-Hye;Park, Eun-Mi;Kim, Young-Ok;Kim, Dong-Gyun;Jee, Young-Ju;Lee, Sang-Jun;An, Cheul Min
    • The Korean Journal of Malacology
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    • v.29 no.3
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    • pp.181-187
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    • 2013
  • In order to investigate environmental stress inducible genes in abalone, we analyzed differentially expressed transcripts from a Pacific abalone, Haliotis discus hannai, after exposure to heat-, cold- or hyposalinity-shock by suppression subtractive hybridization (SSH) method. 1,074 unique sequences from SSH libraries were composed to 115 clusters and 986 singletons, the overall redundancy of the library was 16.3%. From the BLAST search, of the 1,316 ESTs, 998 ESTs (75.8%) were identified as known genes, but 318 clones (24.2%) did not match to any previously described genes. From the comparison results of ESTs pattern of three SSH cDNA libraries, the most abundant EST was different in each SSH library: small heat shock protein p26 (sHSP26) in heat-shock, trypsinogen 2 in cold-shock, and actin in hyposalinity SSH cDNA library. Based on sequence similarities, several response-to-stress genes such as heat shock proteins (HSPs) were identified commonly from the abalone SSH libraries. HSP70 gene was induced by environmental stress regardless of temperature-shock or salinity-stress, while the increase of sHSP26 mRNA expression was not detected in cold-shock but in heat-shock condition. These results suggest that the suppression subtractive hybridization method is an efficient way to isolate differentially expressed gene from the invertebrate environmental stress-response transcriptome.

Differential Protein Expressions in Virus-Infected and Uninfected Trichomonas vaginalis

  • Ding, He;Gong, Pengtao;Yang, Ju;Li, Jianhua;Li, He;Zhang, Guocai;Zhang, Xichen
    • Parasites, Hosts and Diseases
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    • v.55 no.2
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    • pp.121-128
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    • 2017
  • Protozoan viruses may influence the function and pathogenicity of the protozoa. Trichomonas vaginalis is a parasitic protozoan that could contain a double stranded RNA (dsRNA) virus, T. vaginalis virus (TVV). However, there are few reports on the properties of the virus. To further determine variations in protein expression of T. vaginalis, we detected 2 strains of T. vaginalis; the virus-infected ($V^+$) and uninfected ($V^-$) isolates to examine differentially expressed proteins upon TVV infection. Using a stable isotope N-terminal labeling strategy (iTRAQ) on soluble fractions to analyze proteomes, we identified 293 proteins, of which 50 were altered in $V^+$ compared with $V^-$ isolates. The results showed that the expression of 29 proteins was increased, and 21 proteins decreased in $V^+$ isolates. These differentially expressed proteins can be classified into 4 categories: ribosomal proteins, metabolic enzymes, heat shock proteins, and putative uncharacterized proteins. Quantitative PCR was used to detect 4 metabolic processes proteins: glycogen phosphorylase, malate dehydrogenase, triosephosphate isomerase, and glucose-6-phosphate isomerase, which were differentially expressed in $V^+$ and $V^-$ isolates. Our findings suggest that mRNA levels of these genes were consistent with protein expression levels. This study was the first which analyzed protein expression variations upon TVV infection. These observations will provide a basis for future studies concerning the possible roles of these proteins in host-parasite interactions.

Proteomic Analysis of Recombinant Saccharomyces cerevisiae upon Iron Deficiency Induced via Human H-Ferritin Production

  • Seo, Hyang-Yim;Chang, Yu-Jung;Chung, Yun-Jo;Kim, Kyung-Suk
    • Journal of Microbiology and Biotechnology
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    • v.18 no.8
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    • pp.1368-1376
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    • 2008
  • In our previous study, the expression of active H-ferritins in Saccharomyces cerevisiae was found to reduce cell growth and reactive oxygen species (ROS) generation upon exposure to oxidative stress; such expression enhanced that of high-affinity iron transport genes (FET3 and FTR1). The results suggested that the recombinant cells expressing H-ferritins induced cytosolic iron depletion. The present study analyzes metabolic changes under these circumstances via proteomic methods. The YGH2 yeast strain expressing A-ferritin, the YGH2-KG (E62K and H65G) mutant strain, and the YGT control strain were used. Comparative proteomic analysis showed that the synthesis of 34 proteins was at least stimulated in YGH2, whereas the other 37 proteins were repressed. Among these, the 31 major protein spots were analyzed via nano-LC/MS/MS. The increased proteins included major heat-shock proteins and proteins related to endoplasmic reticulum-associated degradation (ERAD). On the other hand, the proteins involved with folate metabolism, purine and methionine biosynthesis, and translation were reduced. In addition, we analyzed the insoluble protein fractions and identified the fragments of Idh1p and Pgk1p, as well as several ribosomal assembly-related proteins. This suggests that intracellular iron depletion induces imperfect translation of proteins. Although the proteins identified above result from changes in iron metabolism (i.e., iron deficiency), definitive evidence for iron-related proteins remains insufficient. Nevertheless, this study is the first to present a molecular model for iron deficiency, and the results may provide valuable information on the regulatory network of iron metabolism.

경구투여 백신 후보물질로서의 Helicobacter pylori 외막 단백질의 조사

  • 박형배;최태부
    • Microbiology and Biotechnology Letters
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    • v.25 no.2
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    • pp.129-136
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    • 1997
  • Helicobacter pylori is a spiral-shaped, microaerophilic human gastric pathogen causing chronic-active gastritis in association with duodenal ulcer and gastric cancer. To investigate the possibility of H. pylori outer membrane proteins (OMPS) as the oral vaccine antigens, sarcosine-insoluble outer membrane fraction has been prepared from H. pylori NCTC 11637. The major OMPs having apparent molecular masses of 62 kDa, 54 kDa and 33 kDa were detected by sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE), which were identified as urease B subunit (UreB), heat shock protein (Hsp54 kDa) and urease A subunit (UreA), respectively. Minor protein bands of 57 kDa, 52 kDa, 40 kDa, 36 kDa and 31 kDa were also observed. The antigenicity of H. pylori OMPs and antigenic cross-reactivity among the strains were determined by immunoblot analysis using anti-H. pylori OMPs antisera or intestinal lavage solutions. The results showed that UreB, Hsp54 kDa, UreA and 40 kDa proteins vigorously stimulated mucosal immune response rather than systemic immunity. From this results, these proteins seemed to be useful as the antigen candidates for the oral vaccine. The immunoblotting results with surface proteins from eight isolated H. pylori strains were similar to that of H. pylori NCTC 11637. The IgA which had been arised from oral administration of H. pylori OMPs, was able to bind H. pylori whole-cells.

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