• 한국동물학회지
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• 제39권2호
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• pp.123-131
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• 1996

In this study, we examined the expression of heat shock proteins (HSPs) in fish cell line CHSE-2lnl and human HeLa cells exposed to heat shock. In fish CHSE-214 cells HSP70 was the major polvpeptide induced by an elevated temperature or an amino acid analog, while in HeLa cells HSP90 as well as HSP70 were prominently enhanced in response to these stresses. Pretreatment of actinomvcin D prior to heat shock completely inhibited the induction of fish HSP70, indicating the transcriptional regulation of fish HSP70 gene expression. In HeLa and CHSE-214 cells either recovering from heat shock or experiencing prolonged heat shock, attenuation in the HSP90 a'nd HSP70 induction occurred but both induction and repression of HSP70 synthesis appear 19 precede those of HSP90. Moreover, attenuation did not occur in the syntheses of 40 kDa and 42 kOto proteins which were only induced in CHSE-214 cells. The enhanced syntheses of these he proteins continued as long as CHSE-214 cells were Siven heat shock. These results suggest that down-regulation of HSP syntheses during prolonged heat shock may be controlled by several different. as vet undefined, mechanisms.

• Parasites, Hosts and Diseases
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• 제59권4호
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• pp.409-413
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• 2021

In this study, we have collected and screened a total of 268 stool samples from diarrheal patients admitted to an Infectious disease hospital in Kolkata for the presence of Cryptosporidium spp. The initial diagnosis was carried out by microscopy followed by genus specific polymerase chain reaction assays based on 70 kDa heat shock proteins (HSP70). DNA sequencing of the amplified locus has been employed for determination of genetic diversity of the local isolates. Out of 268 collected samples, 12 (4.48%) were positive for Cryptosporidium spp. Sequences analysis of 70 kDa heat shock proteins locus in 12 Cryptosporidium local isolates revealed that 2.24% and 1.86% of samples were showing 99% to 100% identity with C. parvum and C. hominis. Along with the other 2 major species one recently described globally distributed pathogenic species Cryptosporidium viatorum has been identified. The HSP70 locus sequence of the isolate showed 100% similarity with a previously described isolate of C. viatorum (Accession No. JX978274.1, JX978273.1, and JN846706.1) present in GenBank.

• Journal of Microbiology and Biotechnology
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• 제10권2호
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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.

• Entomological Research
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• 제48권5호
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• pp.416-428
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• 2018

To gain an insight into the function of heat shock proteins (HSPs) in insects during thermal stress, three HSP cDNAs were identified in the transcriptome of adult Heortia vitessoides, one of the most destructive defoliating pests in Aquilaria sinensis (Loureiro) Sprenger forests. The open reading frames of HvHsp60, HvHsp70, and HvHsp90 were 1,719, 2,070, and 2,151 bp in length, respectively, and encoded proteins with molecular weights of 61.05, 75.02, and 82.23 kDa, respectively. Sequence analysis revealed that all three HSPs were highly conserved in structure. Regarding the stage-specific expression profiles, HvHsp60, HvHsp70, and HvHsp90 mRNAs were detected in all developmental stages. Regarding the tissue-specific expression profiles, the expression levels of the three HSP genes were different in various larval and adult tissues. Moreover, the expression patterns of heat-stressed larvae, pupae, and adults indicated that HvHsp60, HvHsp70, and HvHsp90 were heat-inducible. In particular, HvHsp60 transcripts increased dramatically in larvae and pupae that were heat-stressed at $40^{\circ}C$ and were upregulated in adults that were heat-stressed at $35^{\circ}C$ and $40^{\circ}C$. The expression of HvHsp70 significantly increased in all of the three different developmental stages at $35^{\circ}C$, $40^{\circ}C$, and $45^{\circ}C$. The expression of HvHsp90 obviously increased at $30^{\circ}C$, $35^{\circ}C$, and $40^{\circ}C$ in larvae and could be induced at $35^{\circ}C$ in pupae and adults. The results suggest that HSP60, HSP70, and HSP90 play a major role in protecting H. vitessoides against high-temperature stress.

• 한국어류학회지
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• 제19권4호
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• pp.266-273
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• 2007

열충격 단백질(hsp)은 세포의 기능에 중요한 역할을 하는 보존성이 높은 단백질중의 하나이다. 이들 중 70 kDa 열충격 단백질은 외부의 자극과 관계없이 상시적으로 합성되는 HSC70 단백질과 외부의 자극에 반응하여 합성되는 HSP70 단백질이 있다. 본 연구에서는 넙치(Paralichthys olivaceus)의 70 kDa 열충격 단백질에 대한 cDNA를 아미노산 서열로 변환시켜 분석함으로써 이 유전자가 상시적으로 발현하는 열충격 단백질인 HSC70에 대한 유전자임을 밝혔다. Hsp70 유전자의 발현 기작을 조사하기 위하여 단백질 발현을 조절하는 5' 인접부위를 분리하고 이들의 염기서열을 분석함으로써 유전자 조절부위의 중요인자와 중심 부위를 동정하였다. 또한 Hsp70 유전자의 유전자 조절부위를 이용하여 형광단백질 발현벡터를 제작한 후 메다카 수정란에 미세 주입하여 배 발생 과정의 살아있는 메다카에서 발현하는 형광 단백질(GFP)의 발현을 조사하였다.

• 생물정신의학
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• 제6권2호
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• pp.202-208
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• 1999

Schizophrenia has many clinical expressions and probably different etiologic factors. Infections, autoimmune mechanism and related neurodevelopmental abnormalities have been suggested as possible etiologic factors of schizophrenia. It has been reported that immunoreactivity to heat shock proteins, which play a protective role against environmental stresses in a cell, might be related to the pathogenesis of schizophrenia. Therefore, we examined the immunoreactivity to heat shock protein 70kDa and 90kDa(HSP70 and 90) in 91 patients with schizophrenia and 83 normal controls. Ig G antibodies to HSP70 and 90 of sera were quantitated by ELISA. The optical density(OD) was measured by an automated microplate reader at a wavelength of 490nm. The amounts of antibodies to HSPs were expressed as arbitrary units(AU)/ml related to a standard serum. The limit for elevated antibody titers(anti-HSPs positive or negative) was set at two standard deviations added to the mean of the normal controls. Twenty nine(31.9%) of the 91 patients showed anti-HSP70 positive and 19(20.9%) of those showed anti-HSP90 positive. On the other hand, only 1(1.4%) of the normal controls and 4(4.8%) of those showed anti-HSP70 positive and anti-HSP90 positive, respectively. The titers of anti-HSP70 positive were related with BPRS scores, while those of anti-HSP90 positive were not. There were no relationship between antibody titers and clinical variables including age at onset, duration of illness, family history of schizophrenia or number of admission. The titers of anti-HSP70 positive were significantly associated with anti-HSP90 positive. Our results suggest the presence of abnormal immune reactivity involving HSP70 and HSP90 in a subset of patients with schizophrenia.

• Toxicological Research
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• 제13권1_2호
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• pp.79-86
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• 1997

Inhibitory effects of tannic acid on skin toxicity and heat shock protein induced by UVB were investigated. Tannic acid was administered either topically or orally for 3 days to hairless mice, which were previously irradiated with UVB. UVB was found to cause skin erythema . However, the skin erythema was decreased when tannic acid was administered either topically or orally. The heat shock proteins, Hsp-78 kDa and 70 kDa, were induced by UVB irradiation, but the induction was decreased by treatment of tannic acid in both topically and orally administered groups. The hsp induction was more prominent in orally administered groups than in topically administerd groups. However, the difference between two groups was not statistically significant. The route of administrations, topical and oral, does not affect the activity of tannic acid. In the skin tissue observation, tannic acid regenerated the epithelial cells with 7-9 cell layers which were injured by UVB. In conclusion, tannic acid has an ability to protect against UVB irradiation and regenerate the skin.

• 생명과학회지
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• 제11권5호
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• pp.464-469
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• 2001

내염성의 광합성 cyanobateria 인 Aphanothece halophytica로 부터 molecular chaperone으로 가능하는 HSP70 homolog인 dnaK2 유전자를 cloning 하였다. 이 danK2 유전자는 616개의 아미노산으로 구성되었으며 추정되는 분자량 68 kDa 의 단백질을 code하고 있었다. 아미노산 서열로부터 추정되는 DnaK2 단백질의 구조를 분석하여 본 결과, 다른 원핵생물의 DanK2 단백질들이 공통적으로 갖는 특성인 N-terminal ATPase domain과 C-terminal의 peptide-binding domain이 잘 보존되어 있었으며, 다른 HSP70/DanK 단백질들과의 높은은 상동성을 나타내었다. 한편 danK2 유전자는 생장온도인 28$^{\circ}C$에서 낮은 수준으로 구성적으로 발현하였으며 heat stress에 의해 그 발현량이 급격히 증가하였다. 또한 A. halophytica를 고농도의 염 스트레스로 처리한 결과, heat stress가 없음에도 불구하고 그 발현량이 급격히 증가하였다. 이러한 결과들은 DnaK 단백질의 고온 또는 염 스트레스에 따른 세포의 손상을 보호하기 위하여 중요한 기능을 담당하고 있기 때문에 추정된다.

• Journal of Microbiology and Biotechnology
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• 제19권7호
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• pp.727-733
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• 2009

Heat shock protein 70 kDa (hsp70), a molecular chaperone involved in folding of nascent proteins, has been studied for its ability to activate innate and specific immunity. High purity hsp70 preparation is generally required for immunization experiments, because endotoxins and other immunologically active contaminants may affect immune responses independently of hsp70. We have developed a novel modification of E. coli-expression medium that enabled a simple two-step production and purification method for endotoxin-free recombinant hsp70. During Ni-NTA-based affinity purification of hsp70, a contaminating protein from host E. coli cells, L-glutamine-n-fructose-6-phosphate aminotransferase (GFAT), was identified. By testing various compounds, supplementation of growth medium with a GFAT metabolite,N-acetylglucosamine, was found to reduce GFAT expression and increase the total hsp70 yield five times. The new protocol is based on column purification of His-tagged hsp70 protein produced by E. coli with the modified medium, followed by endotoxin removal by Triton X-114 extraction. This approach yielded hsp70 with high purity and minimal endotoxin contamination, making the final product acceptable for immunization experiments. In summary, a simple modification of growth medium allowed production of recombinant mouse hsp70 in high yield and purity, thus compatible with immunological studies. This protocol may be useful for production of other Histagged proteins expressed in E. coli.

• 환경생물
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• 제20권3호
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• pp.237-244
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• 2002

항진균 활성 관련 단백질을 탐색하기 위해 미역류로부터 식물병원성 곰팡이의 생장을 저해하는 SAR01 균주를 분리하였고, FAME (fatty acid methyl ester) 분석 결과, Streptomyces sp.로 동정되었다. 방사선 조사$(^{60}Co)$를 실시한 결과, Botrytis cinerea를 포함한 5종의 식물병원성 곰팡이에 대한 항진균 활성을 소실한 SAR535 균주 외 6종의 돌연변이 균주가 유도되었다. SAR01 야생형 균주와 SAR535 돌연변이 균주의 세포내 단백질의 이차원 전기영동 분석결과, 6종의 단백질이 야생형 균주인 SAR01 균주의 세포내에만 존재하였다. 이들 6종의 단백질 중, 5종은 heat shock protein 70 (HSP70), Fe-containing superoxide dismutase II (Fe - SODII), ribosome recycling factor (RRF), 10 kD chaperonin (GroES) 및 inorganic pyrophosphatase (PPAse)와 각각 75%, 93%, 100%, 96% 및 83%의 유사성을 보였다. 이들 6종의 단백질들은 Streptomyces sp. SAR01 균주의 항진균 활성과 밀접한 관계가 있을 것으로 사료된다.