• 대한약리학회:학술대회논문집
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• 대한약리학회 2006년도 58th Annual Meeting of The Korean Society of Pharmacology
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• pp.42-42
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• 2006

• 한국동물학회지
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• 제26권3호
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• pp.155-170
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• 1983

溫熱處理에 대해서 細胞가 反應하여 適應하는 過程을 細胞膜水準에서 연구하기 위해서 培養中인 纖維芽細胞性 腫瘍細胞를 이용하여 細胞週期, 細胞密度 및 溫熱處理에 따른 膜表面蛋白質의 變化를 lactoperoxidase를 이용한 iodination과 galactose oxidase를 이용한 tritiation 方法을 서서 分析하였다. 細胞週期와 細胞密度에 따라 膜表面蛋白質은 定量的 變化를 보였는데 $G_1$期에서는 LETS 蛋白質과 高分子蛋白質이 크게 增加하였고 細胞密度의 증가에 따라서는 125K 蛋白質의 增加와 130K 및 100K 蛋白質의 減少가 特異하게 나타났다. 溫熱處理후의 시간경과에 따른 변화를 보면 處理직후 80K 이상의 蛋白質은 모두 사라지고, 24시간 지나면 70K 蛋白質이 현저한 增加를 보였지만 48시간이 경과하면 다시 減少하고 高分子蛋白質들이 原狀으로 回復되었다. 또한, 溫度를 $39^\\circ\\sim45^\\circC$까지 증가시켜 보았을 때 70K 蛋白質이 $41^\\circC$에서 가장 큰 폭으로 增加하는 特異한 현상이 관찰되었다. 아울러 이 70K 단백질은 trypsin을 처리하면 사라졌는데 galactose oxidase로 tritiation하였을때도 iodination하였을 때와 동일한 變化樣相을 보였다. 이러한 결과와 細胞質蛋白質과 比較한 缺課로 미루어 볼 때, 70K 蛋白質은 膜表面蛋白質이며 糖蛋白質이고 또한 HSP 70과 같은 蛋白質인 것으로 推定되었다. 이 蛋白質의 細胞膜에 있어서의 가능한 機能과 腫瘍細胞가 正常細胞에 비해서 溫熱處理에 敏感한 原因등에 관해서 考察하였다.

• Animal Bioscience
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• 제35권1호
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• pp.96-104
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• 2022

Objective: Sarcoplasmic proteins include proteins that play critical roles in biological processes of living organisms. How seasons influence biological processes and meat quality of postmortem muscles through the regulation of protein phosphorylation remain to be investigated. In this study, the phosphorylation of sarcoplasmic proteins in pork longissimus muscle was investigated in four seasons. Methods: Sarcoplasmic proteins were extracted from 40 pork carcasses (10 for each season) and analyzed through ProQ Diamond staining for phosphorylation labeling and Sypro Ruby staining for total protein labeling. The pH of muscle, contents of glycogen and ATP were measured at 45 min, 3 h, and 9 h postmortem and the water (P_2b, P₂₁, and P₂₂) was measured at 3 h and 9 h. Results: A total of 21 bands were detected. Band 8 (heat shock cognate 71 kDa protein; heat shock 70 kDa protein 1B) had higher phosphorylation level in summer than that in other seasons at 45 min postmortem. The phosphorylation levels of 3 Bands were significantly different between fast and normal pH decline groups (p<0.05). The phosphorylation levels of 4 bands showed negative associations with immobilized water (P₂₁) and positive association with free water (P₂₂). Conclusion: The phosphorylation levels of sarcoplasmic proteins involved in energy metabolism and heat stress response at early postmortem time differed depending on the seasons. These proteins include heat shock protein 70, pyruvate kinase, phosphoglucomutase-1, glucose-6-phosphate isomerase, and carbonic anhydrase 3. High temperatures in summer might result in the phosphorylation of those proteins, leading to pH decline and low water holding capacity.

• Bulletin of the Korean Chemical Society
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• 제30권11호
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• pp.2647-2650
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• 2009

Cold shock proteins (Csps) are a subgroup of the cold-induced proteins on reduction of the growth temperature below the physiological temperature. They preferentially bind to single-stranded nucleic acids to translational regulation via RNA chaperoning. Csp plays important role in cold adaptations for the psychrophilic microorganism. Recently, Cold shock protein from psychrophilic bacteria, Colwellia psychrerythraea (CpCsp) has been identified. Three dimensional structures of a number of Csps from various microorganisms have been solved by NMR spectroscopy or X-ray crystallography, but structures of psychrophilic Csps were not studied yet. Therefore, cloning and purification protocols for further structural study of psychrophilic Csp have been optimized in this study. CpCsp was expressed in E. coli with pET-11a vector system and purified by ion exchange, size exclusion, and reverse phase chromatography. Expression and purification of CpCsp in M9 minimal media was carried out and $^{15}N$-labeled proteins with high purity over 90% was obtained. Further study will be carried out to investigate the tertiary structure and dynamics of CpCsp.

• Molecules and Cells
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• 제19권3호
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• pp.328-333
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• 2005

Small heat shock proteins (sHSPs) are widely distributed, and their function and diversity of structure have been much studied in the field of molecular chaperones. In plants, which frequently have to cope with hostile environments, sHSPs are much more abundant and diverse than in other forms of life. In response to high temperature stress, sHSPs of more than twenty kinds can make up more than 1% of soluble plant proteins. We isolated a genomic clone, NtHSP18.3, from Nicotiana tabacum that encodes the complete open reading frame of a cytosolic class I small heat shock protein. To investigate the function of NtHSP18.3 in vitro, it was overproduced in Escherichia coli and purified. The purified NtHSP18.3 had typical molecular chaperone activity as it protected citrate synthase and luciferase from high temperature-induced aggregation. When E. coli celluar proteins were incubated with NtHSP18.3, a large proportion of the proteins remained soluble at temperatures as high as $70^{\circ}C$. Native gel analysis suggested that NtHSP18.3 is a dodecameric oligomer as the form present and showing molecular chaperone activity at the condition tested. Binding of bis-ANS to the oligomers of NtHSP18.3 indicated that exposure of their hydrophobic surfaces increased as the temperature was raised. Taken together, our data suggested that NtHSP18.3 is a molecular chaperone that functions as a dodecameric complex and possibly in a temperature-induced manner.

• 생물정신의학
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• 제14권4호
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• pp.221-231
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• 2007

Recent researches have shown that important cellular-based autoprotective mechanisms are mediated by heat-shock proteins(HSPs), also called 'molecular chaperones'. HSPs as molecular chaperones are the primary cellular defense mechanism against damage to the proteome, initiating refolding of denatured proteins and regulating degradation after severe protein damage. HSPs also modulate multiple events within apoptotic pathways to help sustain cell survival following damaging stimuli. HSPs are induced by almost every type of stresses including physical and psychological stresses. Our nervous system in the brain are more vulnerable to stress and damage than any other tissues due to HSPs insufficiency. The normal function of HSPs is a key factor for endogenous stress adaptation of neural tissues. HSPs play an important role in the process of neurodevelopment, neurodegeneration, and neuroendocrine regulation. The altered function of HSPs would be associated with the development of several neuropsychiatric disorders. Therefore, an understanding of HSPs activities could help to improve autoprotective mechanism of our neural system. This paper will review the literature related to the significance of HSPs in neuropsychiatric field.

• BMB Reports
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• 제38권6호
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• pp.695-702
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• 2005

The groESL bicistronic operon of a restricted facultative methylotrophic bacterium Methylovorus sp. strain SS1 DSM 11726 was cloned and characterized. It was found to consist of two ORFs encoding proteins with molecular masses of 11,395 and 57,396 daltons, which showed a high degree of homology to other bacterial GroES and GroEL proteins. The genes were clustered in the transcription order groES-groEL. Northern blot analyses suggested that the groESL operon is transcribed as a bicistronic 2.2-kb mRNA, the steady-state level of which was markedly increased by temperature elevation. Primer extension analysis demonstrated one potential transcription start site preceding the groESL operon, which is located 100bp upstream of the groES start codon. The transcription start site was preceded by a putative promoter region highly homologous to the consensus sequences of Escherichia coli ${\sigma}^{32}$-type heat shock promoter, which functioned under both normal and heat shock conditions in E. coli. Heat shock mRNA was maximally produced by Methylovorus sp. strain SS1 approximately 10min after increasing the temperature from 30 to $42^{\circ}C$. The groESL operon was also induced by hydrogen peroxide or salt shock.

• 생명과학회지
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• 제26권12호
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• pp.1360-1366
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• 2016

Heat shock proteins (HSPs)은 다양한 생리학적인 또는 환경적 스트레스에 응답하여 유도된다. 그러나 HSPs의 전사 활성은 heat shock factors (HSFs)에 의해 조절 된다. 현재 연구에서는 붕어와 마우스의 간세포 배양에서 열 스트레스에 의한 heat shock factor 1 (HSF1)의 패턴 차이와 heat shock protein 70 (HSP70)의 발현을 면역분석법을 이용하여 조사하였다. 붕어의 간세포는 $33^{\circ}C$에서 trimer를 이루지만 마우스의 간세포는 $42^{\circ}C$에서 trimer를 이루었다. 이 연구는 붕어와 마우스의 HSF1은 열 스트레스로부터 다른 온도에서 반응을 한다는 것을 보여준다. 또한 재조합 단백질을 이용하여 붕어와 인간의 HSF1의 온도에 따른 활성 조건을 CD spectroscopy와 면역분석을 이용하여 조사하였다. 이러한 결과들은 인간과 마우스 HSF1과 붕어의 HSF1은 온도에 의한 활성 변화를 보이지만 그들의 최적 활성 온도는 다르다는 것을 알 수 있다.

• Molecules and Cells
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• 제20권3호
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• pp.378-384
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• 2005

Estrogen metabolites are carcinogenic. The comparative mitogenic activities of $17{\beta}$-estradiol (E2) and four metabolites, 2-hydroxyestradiol (2-OHE2), 4-hydroxyestradiol (4-OHE2), $16{\alpha}$-hydroxyestrone ($16{\alpha}$-OHE1) and 2-methoxyestradiol (2-ME), were determined in estrogen receptor(ER)-positive MCF-7 human breast cancer cells. Each of the E2 metabolites caused proliferation of the MCF-7 cells, but only E2 and $16{\alpha}$-OHE1 induced a greater than 20-fold increases in transcripts of the progesterone receptor (PR) gene, a classical ER-mediated gene. This suggests that the mitogenic action of E2 and $16{\alpha}$-OHE1 could result from their effects on gene expression via the ER. E2 metabolites altered the expression of E2-regulated proteins including heat shock proteins (Hsps). $16{\alpha}$-OHE1 and 2-ME as well as E2 increased levels of Hsp56, Hsp60, $Hsp90{\alpha}$ and Hsp110 transcripts, and the patterns of these inductions resembled that of PR. Hsp56 and Hsp60 protein levels were increased by all the E2 metabolites. Levels of the transcripts of 3 E2-upregulated proteins (XTP3-transactivated protein A, protein disulfide isomerase-associated 4 protein and stathmin 1) and an E2-downregulated protein (aminoacylase 1) were also affected by the E2 metabolites. These results suggest that the altered expression of Hsps (especially Hsp56 and Hsp60) by E2 metabolites such as E2, $16{\alpha}$-OHE1 and 2-ME could be closely linked to their mitogenic action.

• 미생물학회지
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• 제32권1호
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• pp.47-52
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• 1994

Campylobacter jejuni에 48${\circ}C$의 열충격을 30분간 주었을 때, HSP90, HSP66, HSP60의 열충격 단백질들이 합성되었고, 이 단백질들은 각각 E. coli의 hsp87, HSP66 (DnaK), HSP58(GroEL)에 상응하는 단백질들이었다. 여러가지의 제한효소로 처리한 C. jejuni의 chromosomal DNA에 E. coli의 groEL(4.0kb)을 probe로 사용하여 Southern hybridization한 결과, 이들과 상동성을 가지는 유전자들이 있음을 확인하였다. C. jejuni의 groEL 유전자를 pWE15 cosmid를 이용하여 recombinant plasmid pLC1을 만들고, 이를 E. coli B178 groEL44 ts mutant에 형질전환시켜 E. coli LC1을 얻었다. 이 pLC1에는 groEL 유전자가 존재하는 5.7kb인 insert DNA가 포함되어 있었고, 그로부터 subcloning한 pLC101에는 groEL을 포함하는 4.0kb의 DNA가 삽입되어 있었다. 이 recombinant plasmid들이 형질전환된 E. coli LC1과 LC101 균주에서는 C. jejuni의 GroEL 단백질이 과다 생산되었다. C. jejuni의 groEL이 cloning된 E. coli LC1은 42${\circ}C$에서의 생장능력이 회복되었고, ${\lambda}$ vir phage에 대한 감수성도 회복되는 등의 chaperon 효과가 입증되었다.