• 한국초지조사료학회지
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• 제33권3호
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• pp.159-166
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• 2013

We have previously investigated the proteome changes of rice leaves under heat stress (Lee et al. in Proteomics 2007a, 7:3369-3383), wherein a group of antioxidant proteins and heat shock proteins (HSPs) were found to be regulated differently. The present study focuses on the biochemical changes and gene expression profiles of heat shock protein and antioxidant genes in rice leaves in response to heat stress ($42^{\circ}C$) during a wide range of exposure times. The results show that hydrogen peroxide and proline contents increased significantly, suggesting an oxidative burst and osmotic imbalance under heat stress. The mRNA levels of chaperone 60, HSP70, HSP100, chloroplastic HSP26, and mitochondrial small HSP responded rapidly and showed maximum expression after 0.5 or 2 h under heat stress. Transcript levels of ascorbate peroxidase (APX), dehydroascorbate reductase (DHAR) and Cu-Zn superoxide dismutase (Cu-Zn SOD) showed a rapid and marked accumulation upon heat stress. While prolonged exposure to heat stress resulted in increased transcript levels of monodehydroascorbate reductase, peroxidase, glyoxalase 1, glutathione reductase, thioredoxin peroxidase, 2-Cysteine peroxiredoxin, and nucleoside diphosphate kinase 1, while the transcription of catalase was suppressed. Consistent with their changes in gene expression, the enzyme activities of APX and DHAR also increased significantly following exposure to heat stress. These results suggest that oxidative stress is usually caused by heat stress, and plants apply complex HSP- and antioxidant-mediated defense mechanisms to cope with heat stress.

• 생명과학회지
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• 제23권4호
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• pp.586-594
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• 2013

단백질 번역 후 O-GlcNAc 수식은 단백질 조절의 새로운 기전으로 대두되고 있다. 전통적인 당수식과 달리 O-GlcNAc 수식은 단 한번의 O-GlcNAc 전달로 이루어지며, 핵 및 세포질단백질 모두에 수식될 수 있다. O-GlcNAc은 이 분자를 끝으로 하는 최종수식으로 생각되어 왔으나, 최근의 논문(J Proteome Res. 2011 10:2725-2733)은 AP180 단백질에 O-GlcNAc-P가 존재함을 보고하였다. 이 논문은 O-GlcNAc-P가 일반적인 단백질수식인지에 대한 중요한 질문을 던진다. 이에 답하고자 저자들은 HEK293T 세포에 O-GlcNAc 인산화효소 NAGK를 DsRed2에 연결한 DsRed2-$NAGK_{WT}$ 혹은 효소활성이 없는 돌연변이 NAGK를 표현하는 DsRed2-$NAGK_{D107A}$를 표현시키고, 단백질 추출물을 얻어 2D-PAGE로 분리한 후 인산화 정도를 측정하여, $NAGK_{WT}$에 의하여 인산화가 증가되는 15개의 단백질 스폿을 선별하였다. 이 가운데 7개 스팟을 동정한 결과 2개의 스폿은 O-GlcNAc 수식 단백질인 $HSP90{\beta}$, 다른 2개의 스폿도 O-GlcNAc 수식 단백질인 ENO1로 동정되었으며, 나머지(dUTP nucleotidohydrolase mitochondrial isoform 2, glutathione S-transferase P, grp94)는 O-GlcNAc 수식 여부를 아직 모르는 단백질이였다. NAGK에 의하여 O-GlcNAc 단백질의 인산화가 증가된다는 사실은 O-GlcNAc이 인산화되어 O-GlcNAc-P로 수식됨을 시사하며, 따라서 본 연구의 결과는 O-GlcNAc이 최종 수식이 아님을 지지한다.

• KSBB Journal
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• 제32권2호
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• pp.83-89
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• 2017

Gamma (${\gamma}$)-oryzanol is a substance abundant in rice, which is widely cultivated in Asian countries. In this study, we evaluated the effect of ${\gamma}$-oryzanol treatment on proliferation and apoptosis of AGS human gastric carcinoma cells. AGS cells were treated with ${\gamma}$-oryzanol for 72 h in a dose dependent manner. Treatment of ${\gamma}$-oryzanol (50, 100, and $200{\mu}g/mL$) resulted in decreased AGS cell proliferation and increased number of cells in the sub-G1 population. Additionally, apoptotic cells were investigated by annexin V staining and mitochondrial membrane potential assays. Our results indicated that ${\gamma}$-oryzanol treatment increased the number of annexin V-positive cells and depolarized cells. This demonstrated that ${\gamma}$-oryzanol is effective for the induction of apoptosis in AGS cells. We next examined the expression of promising anticancer drug target molecules, including PTEN and HSP90. We found that treatment of ${\gamma}$-oryzanol induced the expression of PTEN in AGS cells. Under the same treatment conditions, ${\gamma}$-oryzanol reduced the expression of HSP90 in AGS cells. These results suggest that ${\gamma}$-oryzanol-induced apoptosis was accompanied by changes in regulation of PTEN and HSP90 in AGS cells. Taken together, ${\gamma}$-oryzanol could be used as a functional substance for the prevention of gastric cancer.

• 생명과학회지
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• 제23권8호
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• pp.1050-1056
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• 2013

Streptococcus pneumoniae는 전 세계적으로 급성 호흡기 질환 높은 사망률 나타내며, 정상인의 비인후부에 존재하여 호흡기 감염을 통해 폐렴, 수막염, 중이염, 패혈증, 복막염, 골수염 등을 일으킨다. 그러나 S. pneumoniae가 폐 조직에 침입하는 분자적 메커니즘과 혈류를 통한 침입은 많은 연구에도 불구하고 아직 명확하게 알려지지 않았다. 그러므로 본 실험에서는 S. pneumoniae D39의 감염 및 침입에 대한 분자 메카니즘을 알고자 사람의 폐암상피 세포 유래 A549 세포를 이용하여 감염 후 시간의 경과에 따라 변화되는 A549 세포의 모양을 관찰하였으며, 또한 숙주세포의 단백질 패턴 변화를 조사하였다. 일부 A549 세포는 감염 후 2 시간부터 세포의 모양이 둥근형태로 변화된 것으로 관찰되었으며, 감염 3 시간째에는 세포의 모양이 둥글며 filopodia가 아주 잘 발달하였다. 감염 4 시간에 도달하게 되면 거의 모든 A549 세포가 둥글며 잘 발달된 filopodia를 형성하였다. 감염 후 각 시간 별 A549 세포의 총 단백질들을 추출하여 시간의 경과에 따라 특이적으로 양 적인 변화를 나타내는 단백질을 MALDI-TOF 분석법을 사용하여 동정하였다. Streptococcus pneumoniae D39 감염 후 시간에 따라 변화하는 단백질 중 대다수가 특이하게도 molecular chaperone에 속하는 단백질들이었다. 대표적인 cytosol chaperone인 Hsp90과 Hsp70의 경우 감소하는 패턴을 나타낸 반면에 endoplasmic reticulum (ER)에 존재하는 chaperone인 Grp94와 Grp78 (BiP)은 감염 후 점차 증가하는 패턴을 나타내었다. ER chaperone인 Grp94와 Grp78의 증가는 ER stress signaling pathway와 관련 있는 것으로 알려져 있어, S. pneumoniae D39의 감염에 의한 이들 단백질의 변화 패턴을 ER stress를 유발 시켰을 때와 비교하였다. Tunicamycin 또는 thapsigargin으로 처리하여 ER stress를 유발시킨 A549 세포의 형태는 변화하지 않았으며 흡착세포의 형태를 유지하였다. 그러나 Western blot을 통한 molecular chaperone의 분석 결과는 S. pneumoniae D39 감염의 경우와 일치하였다. 본 연구에서 얻은 결과는 S. pneumoniae D39의 감염은 A549 세포의 형태적 변화를 유발하며 또한 molecular chaperone 증가와 감소를 유발한다는 것을 보여주며, 특이적으로 Grp94와 Grp78이 증가되는 것으로 보아 S. pneumoniae D39 감염은 A549 세포 내 ER stress를 유발한다고 생각된다.

• Journal of Microbiology
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• 제34권1호
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• pp.37-37
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• 1996

The internal regions of nuclear small subunit rRNA from 6 plaeurotus species and 5 Pleurotus ostreatus strains were amplified by PCR and sequenced. The DNA sequences of 8 Pleurotus strains (P. ostreatus NFFA2, NFFA4501, NFFA4001, KFFA4001, KFCC11635, P florida, P. florida, P. sajor-cuju, P. pulmonarius, and P. spodoleucus) were idential, but P. cornucopiae differed from them in two bases out of 605 bases. However, p[hylogenetic analysis of the sequences by DNA-distance matrix and UPGMA methods showed that P. ostreatus NFFA2m1 and NFFA2m2, known as mutants of P. ostreatus NFFA2, belonged to anther group of Basidiomycotina, which is close to the genus Auricularia. The difference of the SSU rDNA sequences of P. cornucopiae from other Pleurotus species tested corresponds to the difference of mitochondrial plasmid type present in Pleurotus species as observed by Kim et al. (1993, Korean J. Microbiol. 31, 141-147).ishement of silencing at the HMR/hsp82 locus can occur in G1-arrested cells. Cell cycle arrest at G1 phase was achieved by treatment of early log a cell cultures with .alpha.-factor mating pheromone, which induces G1 arrest. The result suggests that passage through S phase (and therefore DNA replication) is nor required for re-establishing silencer-mediated repression at the HMNRa/HSP82 locus. Finally, to test whether de nono protein synthesis is required for re-establishment of silencer-mediated repression, cells were pretreated with cycloheximide (500 /.mu.g/ml) 120 min. It was apparent that inhibiting protein synthesis delays, but does not prevent, re-establishment of silencer-mediated repression. Altogether, these results indicate that re-establishment of silencer-mediated repression is not dependent on the DNA replication and has no requirement for protein synthesis.

• 한국초지조사료학회지
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• 제31권2호
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• pp.99-106
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• 2011

프로테오믹스 기법을 이용하여 벼 고온 스트레스 관련 단백질을 분리 동정하기 위하여 $42^{\circ}C$에서 고온처리한 벼의 줄기로부터 단백질을 분리하였다. 분리한 단백질로부터 Rubisco 단백질을 제거하기 위해 15% PEG fractionation을 실시한 후 상등액 분획의 단백질을 이차원전기 영동한 후, CBB 염색을 통해 차별적 발현을 보이는 단백질을 분석하였다. 총 46개의 단백질 spot이 발현양에 변화를 보였으며, 그 중 24개의 단백질이 고온 스트레스에 의해 발현이 증가되었으며, 22개의 단백질이 감소하는 발현 양상을 나타내었다. 이들 단백질을 MALDI-TOF MS와 database를 통해 동정한 결과 에너지 대사관련 단백질, 산화 환원 관련 단백질 및 저분자량 small HSP 등, 10개의 단백질이 동정되었다. 이들 동정된 단백질들은 식물의 고온 스트레스에 대한 적응기작을 이해하는데 중요한 단서를 제공할 것이며, 특히 미토콘드리아 small HSP는 프로테옴 분석법에 의해 최초로 동정되었으며, 금후 내하고성 목초 분자육종에 활용될 수 있는 좋은 유전자로 판단된다.

• 셀메드
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• 제9권3호
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• pp.4.1-4.8
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• 2019

Now researchers have focused attention on exploring the mechanism of acute responses of heat stress given in heat therapy that ultimately promotes the long term health benefits. Heat therapy is not a new idea rather it was practiced since thousands years back in the form of hot bath, sauna bath, steam room. Similarly in Ayurveda there is very comprehensive description of heat therapy in the form of Svedan karma (Sudation therapy). Svedan is a process to induce sweating artificially in a patient who had already undergone Snehan. Svedan is applied for purification of body, as well as in management of various disorders originated due to vitiation of Vata, Kapha Dosha, Meda Dhatu and musculoskeletal disorders. It produces various beneficial effects by augmenting the Agni like clears the channels, liquefies the deposited Dosha, regulates Vata Dosha, helps in removal and pacification of Dosha, augments metabolism (Agni Deepan), increases appetite, flexibility in body parts, softness and shining of skin, removes coldness, stiffness, drowsiness, improves joint motility. However, Svedana karma is vastly used by Ayurveda Physicians in treatment of various disorders but the mechanisms of beneficial effects produced by Svedan Karma are yet not completely explored on scientific basis. In this article, we will discuss and try to establish a possible mechanism of action of Svedana karma in relation to heat stress, mitochondrial adaptation, heat shock protein (HSP) and glucocorticoids as these are secreted under stressful conditions.

• 대한한방내과학회지
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• 제29권2호
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• pp.469-489
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• 2008

Objectives : The purpose of this study was to investigate the effects of Chungganhaeju-tang(Qingganjiejiu-tang) on alcoholic liver damaged by applying proteomics. Materials and Methods : Sprague-Dawley rats were used in this experiment the rats were divided into the normal group, the control group(alcohol) and the sample group(CGHJT +alcohol). The ethanol was orally administered twice a day for 6 weeks in the control and sample groups. Water instead of ethanol was orally administered twice a day for 6 weeks in the normal group. CGHJT extract was orally administered once a day for 6 weeks in the sample group. The livers of each group were processed and assessed by histology, Western Blot, $Oxyblot^{TM}$, CBB and 2-dimensional electrophoresis. Results : In the histological findings of the liver, CGHJT inhibited hepatic fibrogenesis induced by alcohol. TIMP-1 decreased in the sample group assessed by western blot and statistical significance was noted by dot blotting(p<0.05). In the $Oxyblot^{TM}$, protein oxidation induced by alcohol treatment decreased with CGHJT. In the 2-dimensional electrophoresis finding, increased proteins alcohol such as HSP 60, 60kDa heat shock protein, 3-mercaptopyruvate sulfurtransferase were normalized by CGHJT. CGHJT was considered to normalize the anti-oxidation activity elevated by alcohol. In the 2-dimensional electrophoresis finding, increased oxidized proteins such as actin, prolyl 4-hydroxylase beta polypeptide, 94kDa glucose regulated protein(GRP94), heat shock protein 90-alpha(HSC86), calreticulin precursor(CRP55), ATP synthase beta chain mitochondrial precursor, caspase-8 precursor, and dihydrolipoamide succinyltransferase(E2) decreased with CGHJT. CGHJT was considered to reduce the oxidative stress of alcohol. Conclusion : Chungganhaeju-tang(Qingganjiejiu-tang) exerts an inhibitory effect against the fibrosis and protein oxidation induced by alcohol treatment of rat liver. CGHJT was considered to normalize the elevated anti-oxidation activity by alcohol and to reduce the level of oxidative stress due to alcohol.