• Title/Summary/Keyword: 열충격단백질

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The expression of c-fos and HSP70 by the Capsaicin injection in the spinal cord(dorsal horn) (Capsaicin 적용 후 손상된 흰쥐 척수내 c-fos와 HSP70의 발현)

  • Kim, Dong-Hyun;Kim, Souk-Boum;Baek, Su-Jeong;Kim, Jin-Sang
    • The Journal of Korean Physical Therapy
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    • v.14 no.1
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    • pp.131-137
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    • 2002
  • C-fos는 원종양유전자(proto-oncogene)인 v-fos의 세포 동족체로써, 성장인자나 신경전달 물질에 의해 수분 내에 다양한 형태의 세포에서 활성화된다. Fos단백질은 스트레스와 통증 과정의 신호전달기전에서 세포활동을 조절하는 3차전령으로 활동한다. 열충격 단백질(Heat shock protein : 이하 HSP)은 계통발생학적으로 초기 척추 동물에서부터 발현되며 생체방어체계의 중요한 인자로 세포가 고열, 외상, 허혈 등의 스트레스에 직면했을 때 발현이 증가하는 단백질로 알려져 있다. 본 연구에서는 캡사이신(capsaicin)으로 말초 신경병변을 유발시킨 후 통각신경활성의 지표로 이용되는 원종양 유전자인 c-fos의 발현과 열 또는 스트레스로 야기되는 손상에 대한 조직의 방어작용으로 발현되는 HSP 70의 발현을 동시에 관찰함으로서, 급성으로 유발된 말초 신경병변의 확인과 동시에 실험동물 체내에서 방어적인 역할을 밝히는 일환으로 이 실험을 실시하였다. 본 실험의 결과는 다음과 같다; 1. 척수 등쪽뿔 천층(Laminae I and II)에서 각각 c-fos와 HSP70을 항원으로 하는 면역조직화학적 방법으로 염색한 표본에서 0.9% NaCI 투여 2시간 후 c-fos와 HSP70의 양성을 나타내는 세포는 전혀 없음을 알 수 있었다. 2. 척수 등쪽뿔 천층에서 c-fos 단백질을 항원으로 하는 면역조직화학적 방법으로 염색한 표본에서 Capsaicin 투여 2시간 후 c-fos 단백질에 양성을 나타내는 세포가 많이 발현됨을 육안적 관찰로서 알 수 있었다. 3. 척수 등쪽뿔 천층에서 HSP70을 항원으로 하는 면역조직화확적 방법으로 염색한 표본에서 Capsaicin 투여 2시간 후 HSP7O의 양성을 나타내는 세포가 보통수준으로 발현됨을 육안적 관찰로서 알 수 있었다. 이 실험의 결과로 볼 때, 화학적인 신경병변 유발물질에 의한 손상을 방어하기 위해서 체내에는 내인성 물질이 형성될 것이라는 추측과 c-fos 가 다른 유전자의 발현을 유도한다는 점을 함께 고려 하였을때, Capsaicin에 의한 말초 신경병변에서 c-fos 발현이 많이 나타나는 것은 손상을 방어하는 물질의 생성에 관여하기 때문이며, 방어물질 중 이 실험에서 본 HSP70도 증가한 내인성 방어물질의 하나라고 할 수 있을 것이다.

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Cloning and Immunological Characterization of the 84-kDa Heat Shock Protein, ClpL, in Streptococcus pneumoniae (84-kDa의 폐렴구균 열충격단백질 ClpL의 Cloning 및 면역특성에 관한 연구)

  • 권혁영;김용환;최혜진;박연진;표석능;이동권
    • 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.

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The Effect of Heat Shock Response on the Tumor Necrosis Factor-$\alpha$-induced Acute Lung Injury in Rats (Tumor Necrosis Factor-$\alpha$로 유도되는 백서의 급성 폐손상에 열충격반응이 미치는 효과)

  • Koh, Youn-Suck;Lim, Chae-Man;Kim, Mi-Jung;Cho, Won-Kyung;Jeoung, Byung-O;Song, Kyu-Young;Shim, Tae-Sun;Lee, Sang-Do;Kim, Woo-Sung;Kim, Dong-Soon;Kim, Won-Dong
    • Tuberculosis and Respiratory Diseases
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    • v.44 no.6
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    • pp.1343-1352
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    • 1997
  • Background : Heat-treated cells are known to be protected from lysis by TNF, which is considered to play a central role in the pathogenesis of sepsis-induced acute lung injury. The objective of the study was to investigate the effect of heat shock response by heat-pretreatment on the acute lung injury of the rats induced by intratracheally administered TNF-$\alpha$, Methods : We intratracheally instilled either saline or TNF (R&D, 500ng) with and without heat pretreatment in Sprague-Dawley rats weighing 250~350 g. The heated rats were raised their rectal temperature to $41^{\circ}C$ and was maintained thereafter for 13 minutes at 18 h before intratracheal administration of saline or TNF. After 5 h of intratracheal treatment, lung leak, lung myeloperoxidase activity (MPO) and heat shock proteins were measured in rats. Lung leak index was defined as counts per minute of $I^{25}$ in the right lung divided by counts per minutes of $I^{25}$ in 1.0 ml of blood. All data are expressed as means ${\pm}$SE. Results : There is no difference in acute lung leak index ($0.099{\pm}0.024$ vs $0.123{\pm}0.005$) among the rats given saline intratracheally with and without heat pretreatment, but MPO activity showed a decreased tendency in heat-pretreated rats ($4.58{\pm}0.79\;U/g$) compared with heat-unpretreated rats ($7.32{\pm}0.97\;U/g$) (P=0.064). Rats administered TNF intratracheally with heat-pretreatment had decreased lung leak index ($0.137{\pm}0.012$) and lung MPO activity ($5.51{\pm}1.04\;U/g$) compared with those of heat-unpretreated and TNF-administered rats ($0.186{\pm}0.016$, $14.34{\pm}1.22\;U/g$) (P<0.05 in each). There were no significant difference of lung leak index and MPO activity between TNF-treated rats with heat-pretreatment and saline-treated rats with and without heat-pretreatment. Conclusion : The heat shock response attenuated neutrophil recruitment and acute lung leak induced by intratracheal instillation of TNF-in rats.

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Comparison of Gene Expression in Larval Fat Body of Helicoverpa assulta in Different Temperature Conditions (온도변화에 따른 담배나방 유충 지방체의 유전자 발현 비교 분석)

  • Cha, Wook Hyun;Kim, Kwang Ho;Lee, Dae-Weon
    • Korean journal of applied entomology
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    • v.57 no.3
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    • pp.165-175
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    • 2018
  • Insects are known to live at wide range of temperature, but can not survive when they are exposed to over $40^{\circ}C$ or below supercooling point. The larvae of Helicoverpa assulta have been reared at high ($35^{\circ}C$), low (3 to $10^{\circ}C$), and room temperature ($25^{\circ}C$; control). To identify stress-related genes, the transcriptomes of fat body have been analyzed. Genes such as cuticular proteins, fatty acyl ${\Delta}9$ desaturase and glycerol 3 phosphate dehydrogenase were up-regulated whereas chitin synthase, catalase, and UDP-glycosyltransferase were down-regulated at low temperature. Superoxide dismutase, metallothionein 2, phosphoenolpyruvate carboxykinase and trehalose transporter have been up-regulated at high temperature. In addition, expressions of heat shock protein and glutathione peroxidase were increased at high temperature, but decreased at low temperature. These temperature-specific expressed genes can be available as markers for climate change of insect pests.

T-and cross-reactive B-cell epitopes of Porphyromonas gingivalis and human heat shock protein 60 in atherosclerosis (동맥경화증에 있어서 Porphyromonas gingivalis와 인체 열충격단백의 T-세포 및 교차성 B-세포 epitope)

  • Choi, Jeom-Il
    • Journal of Periodontal and Implant Science
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    • v.33 no.3
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    • pp.331-340
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    • 2003
  • 본 연구의 목적은 인간의 동맥경화증에서 Porphyromonas gingivalis (P. gingivalis)와 인체 열충격단백 60의 T-세포 및 교차성 B-세포 epitope를 규명하고 수립된 T-세포주의 T-세포 주요조직적합체 양상을 파악하려는 데 있다. P. gingivalis 열충격단백-반응성 T 세포주와 환자의 혈청을 이용하여 P. gingivalis 열충격단백60 분자를 구성하는 104개의 중복성 합성 펩타이드의 T-세포 epitope과 B-세포 epitope을 규명하였다. 인체 열충격단백60에 대한 B-세포 epitope도 같은 방법으로 파악하였다. P. gingivalis, P. gingivalis 열충격단백60 또는 인체 열충격단백60에 대한 IgG 항체는 모든 동맥경화증 환자에서 상승하였다. P. gingivalis 열충격단백60의 3, 15, 24, 33, 45, 53, 64, 84, 88, 99번 펩타이드가 주요한 T-세포 epitope였고 이것들은 T-세포 및 B-세포 공동 epitope이기도 했다. 또한 인체 열충격단백60 교차반응 B-세포 epitope은 15, 29, 53, 56, 69, 74번 펩타이드로 판명되었다. 대부분 환자의 주요조직적합체는 $HLA-DRB1^{\ast}1504$$HLA-DZB1^{\ast}0603$으로 나타났다. 결론적으로 P. gingivalis 열충격단백60은 제 2급 주요조직적합제-제한적으로 분해되고 전달되었으며 이 단백질이 공통적인 T-세포 및 B-세포 epitope를 가지면서 동시에 인체 열충격단백60과 교차성 B-세포 epitope을 가지면서 동맥경화증의 면역조절기능에 관여한다고 볼 수 있다.

The Role of DNA Binding Domain in hHSF1 through Redox State (산화환원에 따른 hHSF1의 DNA binding domain의 역할)

  • Kim, Sol;Hwang, Yun-Jeong;Kim, Hee-Eun;Lu, Ming;Kim, An-D-Re;Moon, Ji-Young;Kang, Ho-Sung;Park, Jang-Su
    • Journal of Life Science
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    • v.16 no.6
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    • pp.1052-1059
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    • 2006
  • The heat shock response is induced by environmental stress, pathophysiological state and non-stress conditions and wide spread from bacteria to human. Although translations of most proteins are stopped under a heat shock response, heat shock proteins (HSPs) are produced to protect cell from stress. When heat shock response is induced, conformation of HSF1 was changed from monomer to trimer and HSF1 specifically binds to DNA, which was called a heat shock element(HSE) within the promoter of the heat shock genes. Human HSF1(hHSFl) contains five cysteine(Cys) residues. A thiol group(R-SH) of Cys is a strong nucleophile, the most readily oxidized and nitrosylated in amino acid chain. This consideration suggests that Cys residues may regulate the change of conformation and the activity of hHSF1 through a redox-dependent thiol/disulfide exchange reaction. We want to construct role of five Cys residues of hHSF by redox reagents. According to two studies, Cys residues are related to trimer formation of hHSF1. In this study, we want to demonstrate the correlation between structural change and DNA-binding activity of HSF1 through forming disulfide bond and trimerization. In this results, we could deduce that DNA binding activity of DNA binding domain wasn't affected by redox for always expose outside to easily bind to DNA. DNA binding activity of wild-type HSF's DNA binding domain was affected by conformational change, as conformational structure change (trimerization) caused DNA binding domain.

Effect of Arsenic on Heat Shock Protein and Vascular Contractility of Rat Aorta (횐쥐 대동맥의 수축반응과 열충격단백질에 대한 비소의 영향)

  • 박태규;권윤정;김중영
    • Journal of Environmental Science International
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    • v.12 no.6
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    • pp.651-657
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    • 2003
  • In order to examine if arsenic, one of environmental stresses, contributes to hypertension as one of cardiovas cular pathological factors, this study was perfarmed in vivo and in vitro, using intacted or pithed rats and aorta ring preparation, respectively. And also the relationship between expression of heat shock protein (HSP) 90 and vasoactives-induced contractile response was elucidated. To measure blood pressure, the carotid arterial pressure was recorded on physiograph(Grass Co. 79E) connected to strain gauge. On the other hand, contractile response of vascular ring preparation isolated from rat was determined in organ bath and was recorded on physiograph connected to isometric transducer. And HSP was detacted by Western blotting whole cell Iysis. Preganglionic nerve stimulation was increased by 26.0% in arterial pressure of rat treated with arsenic. Vascular contractile response was monitored and HSP were measured by Western blotting of whole Iysis prepared from samples exposed with 0, 0.5, 1, 2 and 4 mM of arsenic for 8 hours. The dose-vascular responses of potassium chloride were augmented by increasing dose of arsenic in the strips exposed to arsenic for 8 hours, and were not augmented for 1, 3, 5 hours. And the response of relaxation of rat aorta induced by histamine was not influenced by arsenic stress. The increase of HSP 90 expression in rat aorta was pronounced at 8 hours after 4 mM of arsenic treatment, but HSP 60 expression was not. Arsenic stress not only increased the expression of HSP 90 in the rat aorta, but also augmented contractions to potassium chloride. These results indicated that arsenic stress was sufficient to induce heat shock protein 90, resulting in increased vascular contractility in rat aorta.

Differential Susceptibility to High Temperature and Variation of Seasonal Occurrence between Spodoptera exigua and Plutella xylostella (파밤나방과 배추좀나방의 고온 감수성 차이와 연중 발생 변이)

  • Kim, Minhyun;Lee, Seunghee;Kim, Yonggyun
    • Korean journal of applied entomology
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    • v.55 no.1
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    • pp.17-26
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    • 2016
  • Climate change has been regarded as one of main factors to change Korean insect pest fauna. Especially, a global warming model predicts to expand habitat for insect pests originated from tropical or subtropical regions. Two insect pests, the beet armyworm (Spodoptera exigua) and the diamondback moth (Plutella xylostella), are known to overwinter in some greenhouse conditions without diapause induction in Korea. There was a clear difference between these two insects in seasonal occurrence. P. xylostella occurred only at early spring and fall seasons, but did not occur during summer. In contrast, S. exigua maintained their occurrence from late spring to fall seasons. This study set up a hypothesis that the difference in the seasonal occurrence may be resulted from variation in susceptibility to high temperature. To test the hypothesis, heat tolerance was compared between these two insects. Exposure to $42^{\circ}C$ for 40 min killed 100% individuals of P. xylostella larvae. However, most larvae of S. exigua survived in response to $42^{\circ}C$ even for 80 min. Heat tolerance varied among developmental stages in both insects. Highest tolerant stages were $4^{th}$ instar larvae and adults for P. xylostella, but $1^{st}$ instar larvae for S. exigua. Pre-exposure to $37^{\circ}C$ for 30 min significantly increased heat tolerance in both insects. Induction of heat tolerance accompanied with significant increase of glycerol contents in the hemolymph in both insects and up-regulation of three heat shock protein expressions in S. exigua. These results suggest that the differential susceptibility to high temperature explains the disappearance of P. xylostella during summer, at which S. exigua maintains its occurrence.

Characteristics of Soybean Soaking Water after Heat Treatment (대두 열처리 담금 용액의 특성)

  • Yoon, Hye-Hyun;Jeon, Eun-Jae
    • Korean Journal of Food Science and Technology
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    • v.35 no.6
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    • pp.1098-1103
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    • 2003
  • Soybeans released proteins when immersed in water at $50{\sim}60^{\sim}C$. We investigated the changes in the characteristics of soybean when soaked in water at different temperatures and studied the electrophoretic properties of soy proteins in recommended Korean soybean varieties after heat treatment. Soybean seeds were heated in soaking water at temperatures of 30, 40, 50, 60, $70^{\circ}C$ for 90 min, and also from 10 to 150min at $60^{\circ}C$. The pH value of the water decreased with heating time at $60^{\circ}C$, and the amount of soluble solids increased with temperature and heating time. The protein concentration of the solution increased with temperature and time. From SDS-PAGE of the proteins in soaking water, we detected two new bands of 16 kDa- and 31 kDa-proteins from the Korean soybean varieties on heat treatment.

Cold Hardiness Change in Solenopsis japonica (Hymenoptera: Formicidae) by Rapid Cold Hardening (급속내한성 유기에 의한 일본열마디개미(Solenopsis japonica)의 내한성 변화)

  • Park, Youngjin;Vatanparast, Mohammad;Lee, Jieun
    • Korean journal of applied entomology
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    • v.60 no.2
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    • pp.193-199
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    • 2021
  • Solenopsis japonica, which is belonging to Formicidae in Hymenoptera, is a native ant species in Korea. However, it had not been studied for cold hardiness of S. japonica to understand on its overwintering mechanisms in field so far. Cold tolerance on developmental stages was measured at different cold temperature with various exposure times. Workers showed more survival at 5℃ and 10℃ compared with other stages and elevated cold tolerance when workers were exposed at 15℃ for more than 12h incubation as a rapid cold hardening (RCH) condition. RCH treatment not only increased survival of workers at cold temperatures, but also decreased supercooling point (SCP) and freezing point (FP). RCH group increased the survival rate by 44% at 10℃ compared with Non-RCH group. SCP and FP were depressed from -10.0 to -14.2℃ and from -11.3 to -15.3℃, respectively, after RCH treatment. Cold temperature increased expression level of cold- and stress-related genes such as glycerol kinase and heat shock protein. These results indicate unacclimated cold tolerance of S. japonica and its acclimation to low temperature by RCH.