• Molecules and Cells
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• v.28 no.3
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• pp.175-181
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• 2009

In hypertriglyceridaemic individuals, atherosclerogenesis is associated with the increased concentrations of very low density lipoprotein (VLDL) and VLDL-associated remnant particles. In vitro studies have suggested that VLDL induces foam cells formation. To reveal the changes of the proteins expression in the process of foam cells formation induced by VLDL, we performed a proteomic analysis of the foam cells based on the stimulation of differentiated THP-1 cells with VLDL. Using two-dimensional gel electrophoresis (2-DE) and matrix-assisted laser-desorption ionization time-of-flight mass spectrometry (MALDI-TOF MS) analysis, 14 differentially expressed proteins, containing 8 up-regulated proteins and 6 down-regulated proteins were identified. The proteins are involved in energy metabolism, oxidative stress, cell growth, differentiation and apoptosis, such as adipose differentiation-related protein (ADRP), enolase, S100A11, heat shock protein 27 and so on. In addition, the expression of some selected proteins was confirmed by Western blot and RT-PCR analysis. The results suggest that VLDL not only induces lipid accumulation, but also brings about foam cells diverse characteristics by altering the expression of various proteins.

• Journal of Animal Science and Technology
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• v.57 no.12
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• pp.44.1-44.9
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• 2015

Background: Heat shock proteins play an important role in protection from stress stimuli and metabolic insults in almost all organisms. Methods: In this study, computational tools were used to deeply analyse the physicochemical characteristics and, using homology modelling, reliably predict the tertiary structure of the blunt snout bream (Ma-) Hsp70 and Hsc70 proteins. Derived three-dimensional models were then used to predict the function of the proteins. Results: Previously published predictions regarding the protein length, molecular weight, theoretical isoelectric point and total number of positive and negative residues were corroborated. Among the new findings are: the extinction coefficient (33725/33350 and 35090/34840 - Ma-Hsp70/ Ma-Hsc70, respectively), instability index (33.68/35.56 - both stable), aliphatic index (83.44/80.23 - both very stable), half-life estimates (both relatively stable), grand average of hydropathicity (-0.431/-0.473 - both hydrophilic) and amino acid composition (alanine-lysine-glycine/glycine-lysine-aspartic acid were the most abundant, no disulphide bonds, the N-terminal of both proteins was methionine). Homology modelling was performed by SWISS-MODEL program and the proposed model was evaluated as highly reliable based on PROCHECK's Ramachandran plot, ERRAT, PROVE, Verify 3D, ProQ and ProSA analyses. Conclusions: The research revealed a high structural similarity to Hsp70 and Hsc70 proteins from several taxonomically distant animal species, corroborating a remarkably high level of evolutionary conservation among the members of this protein family. Functional annotation based on structural similarity provides a reliable additional indirect evidence for a high level of functional conservation of these two genes/proteins in blunt snout bream, but it is not sensitive enough to functionally distinguish the two isoforms.

• Molecules and Cells
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• v.25 no.1
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• pp.55-63
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• 2008

Amyotrophic lateral sclerosis (ALS) is a progressive neurodegenerative disorder characterized by the selective death of motor neurons. Mutations in the SOD1 gene are responsible for a familial form of ALS (FALS). Although many studies suggest that mutant SOD1 proteins are cytotoxic, the mechanism is not fully understood. To investigate the role of mutant SOD1 in FALS, human SOD1 genes were fused with a PEP-1 peptide in a bacterial expression vector to produce in-frame PEP-1-SOD fusion proteins (wild type and mutants). The expressed and purified PEP-1-SOD fusion proteins were efficiently transduced into neuronal cells. Neurones harboring the A4V, G93A, G85R, and D90A mutants of PEP-1-SOD were more vulnerable to oxidative stress induced by paraquat than those harboring wild-type proteins. Moreover, neurones harboring the mutant SOD proteins had lower heat shock protein (Hsp) expression levels than those harboring wild-type SOD. The effects of the transduced SOD1 fusion proteins may provide an explanation for the association of SOD1 with FALS, and Hsps could be candidate agents for the treatment of ALS.

• Journal of Microbiology and Biotechnology
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• v.16 no.3
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• pp.368-373
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• 2006

Bacillus subtilis and related Bacillus species are frequently used as hosts for the mass production of recombinant proteins. Accordingly, this study examined the cellular response of B. subtilis to the overexpression of a soluble secretory protein. As such, the lichenase derived from B. cereus was overexpressed in B. subtilis, initially localized in the cytoplasm as a mature form and then secreted into the medium. Thereafter, the proteome of B. subtilis was analyzed using 2D electrophoresis and MALDI-TOF mass spectrometry. The expression of several heat-shock proteins, such as dnaK and groEL, was increased under this condition. In addition, manganese superoxide dismutase and NADH dehydrogenase were also upregulated in the lichenase-secreting B. subtilis. Therefore, it was concluded that the transient accumulation of a secreted protein in B. subtilis before secretion acted as a stress on the cell, which in turn induced the expression of various protective proteins.

• 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.

• Journal of Life Science
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• v.23 no.8
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• pp.1050-1056
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• 2013

Streptococcus pneumoniae is the leading cause of community-acquired pneumonia throughout the world. The bacteria invade through lung tissue and cause sepsis, shock, and serious sequelae, including rheumatic fever and acute glomerulonephritis. However, the molecular mechanism associated with pneumonia's penetration of lung tissue and invasion of the blood stream are still unclear. We attempted to investigate the host cell response at protein levels to S. pneumoniae D39 invasion using human lung cancer epithelial cells, A549. Streptococcus pneumoniae D39 began to change the morphology of A549 cells to become round with filopodia at 2 hours post-infection. A549 cell proteins obtained at each infection time point were separated by SDS-PAGE and analyzed using MALDI-TOF. We identified several endoplasmic reticulum (ER) resident proteins such as Grp94 and Grp78 and mitochondrial proteins such as ATP synthase and Hsp60 that increased after S. pneumoniae D39 infection. Cytosolic Hsc70 and Hsp90 were, however, identified to decrease. These proteins were also confirmed by Western blot analysis. The identified ER resident proteins were known to be induced during ER stress signaling. These/ data, therefore, suggest that S. pneumoniae D39 infection may induce ER stress.

• Proceedings of the Microbiological Society of Korea Conference
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• 2002.10a
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• pp.76-78
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• 2002

Heat shock proteins (HSPs) are induced in most living cells by mild heat treatment, ethanol, heavy metal ions and hypoxia. In yeast Saccharomyces cerevisiae, mild heat pretreatment strongly induces Hsp104 and thus provide acquired thermotolerance. The ability of hsp104 deleted mutant $({\triangle}hsp104)$ to acquire tolerance to extreme temperature is severely impaired. In providing thermotolerance, two ATP binding domains are indispensible, as demonstrated in ClpA and ClpB proteases of E. coli. The mechanisms by which Hsp104 protects cells from severe heat stress are not yet completely elucidated. We have investigated regulation of mitochondrial metabolic pathways controlled by the functional Hsp104 protein using $^{13}C_NMR$ spectroscopy and observed that the turnover rate of TCA cycle was enhanced in the absence of Hsp104. Production of ROS, which are toxic to kill cells radiply via oxidative stress, was also examined by fluorescence assay. Mitochondrial dysfunction was manifested in increased ROS levels and higher sensitivity for oxidative stress in the absence of Hsp104 protein expressed. Finally, we have identified mitochondrial complex I and Ferritin as binding protein(s) of Hsp104 by yeast two hybrid experiment. Based on these observations, we suggest that Hsp104 protein functions as a protector of oxidative stress via either keeping mitochondrial integrity, direct binding to mitochonrial components or regulating metal-catalyzed redox chemistry.

• Asian-Australasian Journal of Animal Sciences
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• v.23 no.4
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• pp.437-443
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• 2010

To be economically profitable, the poultry industry demands an increase in stocking density, which could adversely affect chicken welfare. The current study was performed to investigate the effect of stocking density on stress-related, heat shock protein genes (HSP70 and HSP90), 3-hydroxyl-3-methyl-glutaryl coenzyme A reductase (HMGCR) gene and telomere length in broiler chickens. Seven-day-old broiler chickens were housed at High (0.0578 $m^2$/bird), Standard (0.077 $m^2$/bird) and Low (0.116 $m^2$/bird) stocking densities with 8 replicates each until 35 d of age. The growth performance, such as body weight gain and average daily feed intake, was found to be significantly (p<0.05) higher in the Low density group, but these parameters did not show any difference between the High and Standard groups. Other growth performance, such as feed conversion ratio and final feed intake, showed no difference among the treated groups. The expression levels of HSP70 and HMGCR were found to be elevated with the increase of stocking density. The expression level of these genes was significantly (p<0.05) higher in the High density stocked group compared with the other groups, whereas the expression levels were not significantly different between the Low and Standard groups. The expression levels of HSP90 did not show any significant changes among the treated groups. The telomeric length of the birds housed in High density was reduced significantly (p<0.05) when compared to that of the birds in Low density. These results clearly indicate that birds stocked at high density show physiological adaptive changes indicative of stress at gene transcriptional and telomere levels.

• Asian-Australasian Journal of Animal Sciences
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• v.25 no.8
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• pp.1096-1101
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• 2012

Intrauterine growth retardation (IUGR) leads to the dysfunction in digestive system, as well as the alteration in the expression of some functional proteins. Heat shock protein 70 (Hsp70) could be induced by various stress factors, but whether Hsp70 expression is changed in neonatal IUGR infants has not been demonstrated. This study was conducted to explore the expression of Hsp70 in the liver by using the IUGR piglet model. Liver and plasma samples were obtained from IUGR and normal birth weight (NBW) piglets at birth. The neonatal IUGR piglets had significantly lower liver weight than their counterparts. The activities of aspartate aminotransferase and alanine aminotransferase in serum were enhanced significantly in IUGR indicating liver dysfunction. The activities of superoxide dismutase (p<0.01), glutathione peroxidase (p<0.01) and catalase (p>0.05) were lower and the level of malondialdehybe was higher (p<0.05) in IUGR liver compared with in NBW. According to the results of histological tests, fatty hepatic infiltrates and cytoplasmic vacuolization were present in the liver of IUGR piglets, but not in NBW liver. The expression of Hsp70 protein was significantly higher (p<0.05) in IUGR piglet liver than in NBW. Similar to where the hepatic injuries were observed, location of Hsp70 was mostly in the midzonal hepatic lobule indicating that oxidative stress might be responsible for the increased expression of Hsp70.

• Journal of the Society of Cosmetic Scientists of Korea
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• v.30 no.4 s.48
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• pp.471-477
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• 2004

Exposure to elevated temperatures, chemical (active oxigen), or physical stress (UV light) induces immediate physiological response, the expression of heat shock proteins in cells. Thus, cells with elevated Heat Shock Protein levels become more tolerant to stress conditions that are otherwise lethal. First, we studied on the new function of glucuronic acid (GA) as preventive material of skin aging. The application of the GA shows significant induction of Heat Shock Protein 70 kDa (HSP 70 kDa) in contrast to cells without it. GA at the concentration which can induce HSP 70 kDa, protects the cell death induced by second stress (heat shock and hydrogen peroxide) in NIH3T3 cells. Second, we studied on in vitro transdermal permeation characteristic of GA through the excised mouse skin. In this study, we compared the skin permeability of GA in water with O/W emulsion. As a result, skin permeation parameters of GA shows lag time 1.2 h, partition coefficient 0.114, permeation flult rate $0.83114 mg/cm^2/h.$ In case of lag time, O/W emulsion containing GA increase 2.48 h. Also, the total accumulation permeation content decreased in contrast to GA solution after 24 h. But it has long-term permeability of glucuronic acid. These results suggest that glucuronic acid could be a good cosmetic active ingredient.