• 미생물학회지
• /
• 제45권4호
• /
• pp.291-299
• /
• 2009

본 연구에서는 Human HtrA3 (HtrA3)의 효소활성을 분자수준에서 연구하기 위해 HtrA간의 상동성과 기존에 알려진 maturation site들을 비교 분석하여 예상 mature HtrA3인 M1-HtrA3와 M2-HtrA3를 발현하는 construct를 제작하였다. pGEX system을 통해 Top10 균주에서 발현, 정제한 M1-HtrA3 단백질은 $10{\mu}g$/L를 정제할 수 있었으며 발현량 대비 1%를 회수할 수 있었다. M2-HtrA3는 M1보다 5배 가량 많은 양을 정제할 수 있었으며 발현량 대비 회수율은 3배 정도 더 높았다. $\beta$-Casein을 이용한 in vitro cleavage test를 통해 M1, M2 form 모두 protease 활성을 갖는 것을 확인하였다. 또한, $\beta$-casein cleavage를 통해 HtrA serein protease들 간의 상대적인 활성을 비교한 결과, HtrA3와 HtrA2는 HtrA1보다 약 2배 더 높은 proteolytic cleavage 활성을 보였다. 본 연구에서 정립한 protease 활성을 지닌 HtrA3의 제작과 정제 조건은 HtrA3의 substrate를 탐색을 용이하게 할 수 있을 것이며, HtrA3 연관된 질환의 발병기전과 세포 신호전달을 이해하는 연구에 활용될 수 있을 것이다.

• 한국전통조경학회지
• /
• 제34권3호
• /
• pp.1-17
• /
• 2016

본 연구는 고종 연간 경복궁 후원의 북쪽에 조성된 오운각 권역의 특성과 창덕궁 후원 옥류천에 구비되어 있던 기능적 요소가 경복궁 후원으로 치환되었던 양상을 고찰한 것이다. 오운각 권역은 샘과 천하제일복지 각자가 있던 기존의 공간을 활용하여 왕의사적인 공간으로 정비된 공간이었다. 고종은 오랫동안 창덕궁 옥류천이 담당했던 왕실의 휴식과 위락기능을 중건 경복궁에 그대로 옮기려 하였다. 그 결과, 옥류천과 유사한 기능을 하도록 조성된 오운각 권역이 후원의 별원으로 개발되었다. 양자 간에는 후원의 최북단에 조성된 입지 외에도, 건물의 구성 및 유배거의 기능을 했던 것으로 추정되는 관상용 수경설계 등에서 공통점이 발견되었다. 오운각 권역은 옥련정, 오운각, 벽화실과 천하제일복지천의 수경시설로 구성되었다. 이중에서 벽화실을 제외한 나머지는 모두 옥류천과 직간접적인 관련성을 갖되, 지형의 굴곡, 경사, 수계의 상황 등 주어진 입지조건에 따라 조성되어 색다른 정감을 갖는 공간으로 개발되었다. 먼저, 옥련정은 남산에 이르는 서울의 경관을 조망하기 위해 높은 언덕에 입지하였고, 천하제일복지천의 독특한 형식의 수로는 위이암의 유배거의 기능을 모방한 형태로 추정된다. 한편 오운각은 옥류천의 농산정과 같이 주거가 가능한 집으로 조영된 것이었다. 이러한 구성요소는 다분히 옥류천의 기능과 위상을 경복궁에 치환하기 위한 조성목적을 표출하고 있으며, 이러한 점은 조선시대 궁궐 후원의 기능 공간이었던 별원의 특성을 잘 대변하는 것으로 해석되었다.

• The Korean Journal of Physiology and Pharmacology
• /
• 제12권5호
• /
• pp.245-251
• /
• 2008

Single-channel recordings of TASK-1 and TASK-3, members of two-pore domain $K^+$ channel family, have not yet been reported in dorsal root ganglion (DRG) neurons, even though their mRNA and activity in whole-cell currents have been detected in these neurons. Here, we report single-channel kinetics of the TASK-3-like $K^+$ channel in DRG neurons and up-regulation of TASK-3 mRNA expression in tissues isolated from animals with spinal cord injury (SCI). In DRG neurons, the single-channel conductance of TASK-3-like $K^+$ channel was $33.0{\pm}0.1$ pS at - 60 mV, and TASK-3 activity fell by $65{\pm}5%$ when the extracellular pH was changed from 7.3 to 6.3, indicating that the DRG $K^+$ channel is similar to cloned TASK-3 channel. TASK-3 mRNA and protein levels in brain, spinal cord, and DRG were significantly higher in injured animals than in sham-operated ones. These results indicate that TASK-3 channels are expressed and functional in DRG neurons and the expression level is up-regulated following SCI, and suggest that TASK-3 channel could act as a potential background $K^+$ channel under SCI-induced acidic condition.

• 대한수의학회지
• /
• 제51권2호
• /
• pp.139-149
• /
• 2011

Experimental autoimmune encephalomyelitis (EAE) is an inflammatory disease in the murine central nervous system (CNS) and has long been used as an animal model for human multiple sclerosis. Development of EAE requires coordinated expression of a number of genes that are involved in the activation and effector functions of inflammatory cells. Galectin-3 (Gal-3) is a member of the betagalactoside- binding lectin family and plays an important role in inflammatory responses through its functions on cell activation, cell migration or inhibition of apoptosis. We investigated the functional role of Gal-3 in EAE mice following immunization with myelin oligodendrocyte glycoprotein $(MOG)_{35-55}$ peptide. During the peak stage of EAE, the localization of Gal-3 in inflammatory cells markedly increased in subarachnoid membranes and perivascular regions of CNS. In contrast, Gal-3 was weakly detected in cerebrum and spinal of the recovery stage of EAE. Consistent with this finding, western blot analysis revealed that Gal-3 expression was significantly increased at the peak stage while it was slightly decreased at the recovery stage in the CNS. In addition, the population of $CD11b^{+}$ macrophage expressing Gal- 3 in spleen of EAE mice was markedly increased compared with control mice. In fact, most of activated macrophages isolated from spleen of EAE mice expressed Gal-3. Taken together, our results demonstrate that the over-expression of Gal-3 in activated macrophages may play a key role in promoting inflammatory cells in the CNS during EAE.

• 미생물학회지
• /
• 제47권4호
• /
• pp.295-301
• /
• 2011

아쿠아포린(aquaporin)은 MIP (Major Intrinsic Protein) 패밀리에 속하는 물 수송 채널(water transport channel) 단백질로 단세포 생물인 박테리아부터 다세포 고등생물인 인간에 이르기까지 다양한 기관계에서 잘 보존되어 있다. 아쿠아포린은 정통아쿠아포린(orthodox aquaporin)과 아쿠아글리세로포린(aquaglyceroporin)으로 구분되는데, 정통아쿠아포린은 주로 세포내의 물 유입 및 수송에 관여하며 아쿠아글리세로포린은 glycerol, polyol, urea를 비롯한 작은 비극성 분자의 수송에 관여하는 것으로 알려져 있다. 최근까지 효모에서 아쿠아포린 기능이 일부 밝혀졌지만 Aspergillus 속을 포함하는 사상성 진균에서는 거의 연구가 되어있지 않은 실정이다. 본 연구에서는 A. nidulans의 유전체 염기서열 정보를 분석하여 하나의 정통아쿠아포린(aqpA)과 네 개의 아쿠아글리세로포린(aqpB-E)을 발견하였다. 이를 바탕으로 aqpA 유전자 결실돌연변이들을 만들어 그 기능을 분석하였다. aqpA 결실돌연변이는 각종 삼투 스트레스(osmotic stress)에서는 표현형의 변화가 거의 관찰되지 않았으며 이는 이들 유전자가 삼투 스트레스에 반응하지 않거나 유전자의 중복성 때문으로 여겨진다. 그러나 항진균제인 fluconazol에 대해서 그 감수성이 적어지는 것이 관찰 되었다. 이는 aqpA 유전자가 삼투스트레스 반응보다 항진균제의 감지에 더 기능을 가지고 있을 수 있음을 시사한다.

• Molecules and Cells
• /
• 제27권4호
• /
• pp.467-473
• /
• 2009

Our previous study suggested that OsBWMK1, a gene which encodes a member of the rice MAP kinase family, generates transcript variants which show distinct expression patterns in response to environmental stresses. The transcript variants are generated by alternative splicing and by use of alternative promoters. To test whether the two alternative promoters, pOsBWMK1L (promoter for the OsBWMK1L splice variant) and pOsBWMK1S (promoter for the OsBWMK1S splice variant), are biologically functional, we analyzed transgenic plants expressing GUS fusion constructs for each promoter. Both pOsBWMK1L and pOsBWMK1S are biologically active, although the activity of pOsBWMK1S is lower than that of pOsBWMK1L. Histochemical analysis revealed that pOsBWMK1L is constitutively active in most tissues at various developmental stages in rice and Arabidopsis, whereas pOsBWMK1S activity is spatially and temporally restricted. Furthermore, the expression of pOsBWMK1S::GUS was upregulated in response to hydrogen peroxide, a plant defense signaling molecule, in both plant species. These results suggest that the differential expression of OsBWMK1 splice variants is the result of alternative promoter usage and, moreover, that the mechanisms controlling OsBWMK1 gene expression are conserved in both monocot and dicot plants.

• 유통과학연구
• /
• 제12권8호
• /
• pp.103-112
• /
• 2014

Purpose - There have been local government systems since June 1955. Many local governments have made use of festivals for regional public relations and the achievement of competitiveness. This study analyzes effective components of local festivals. There has been extensive research in relation to variables such as participation motivation, satisfaction level, recall intention, royalty, and recommend purpose. However, as a variable factor, attachment to the participation motivation of a festival has been overlooked along with support level to a festival. This study has added new variables named "attachment to local society" and "support level," which are dependent variables. We will study the relationship of these two variables. Research design, data, and methodology - This study has devised a cause-and-effect relationship model, which deduces that local residents' participation motivation for the festival will have positive influence on the festival support level and satisfaction level. We circulated questionnaires to local residents to get the data required to prove the hypothesis. To analysis the data, we used SPSS 21.0, and accomplished basic analysis through the analysis of frequency as well as descriptive statistical analysis. Further, we verified our measurement tools through reliability analysis, and examined the research hypothesis through multiple regression analysis. Results - With regard to the festival satisfaction level, the variables of local society attachment, festival attraction, escaping from the daily life, and family harmony have influence upon a positive effect (+) in this order. Among them, the variable of local society attachment has the greatest influence. However, in respect of support level for the festival, only the variables of local society attachment and escaping from the daily life have a valid influence on the positive effect (+), and local society attachment has relatively greater influence. In addition, not only does the festival satisfaction level affect the support level for the festival in terms of positive effect, the influence is also far-reaching, which verifies the mediating effect. Conclusions - The result shows that the local residents' satisfaction level with the Yudalsan Mountain Spring Flower Festival is comparatively high, which is due to the local society attachment. It also shows that it is difficult to create support without local residents' attachment with their area. Consequently, this study insists that the Yudalsan Mountain Spring Flower Festival should exceed its limitation as just a village-level party, in order to become a remarkable and outstanding festival, and suggests three operational implications: macro-environmental development, task environment development, and internal operational environmental development. From the macro-environmental perspective, the constant development of local culture and local business is proposed. In the task environment perspective, a new functional development for the festival is proposed. From the internal operational environmental perspective, improvements in festival operation methods are proposed. If these weak points are addressed and resolved through a more detailed approach, it will be very helpful for local government systems to operate the festival.

• Experimental and Molecular Medicine
• /
• 제50권11호
• /
• pp.8.1-8.14
• /
• 2018

We previously demonstrated that the direct transplantation of human umbilical cord blood-derived mesenchymal stem cells (hUCB-MSCs) into the dentate gyrus ameliorated the neurological symptoms of Niemann-Pick type C1 (NPC1)-mutant mice. However, the clinical presentation of NPC1-mutant mice was not fully understood with a molecular mechanism. Here, we found 14,15-epoxyeicosatrienoic acid (14,15-EET), a cytochrome P450 (CYP) metabolite, from hUCB-MSCs and the cerebella of NPC1-mutant mice and investigated the functional consequence of this metabolite. Our screening of the CYP2J family indicated a dysregulation in the CYP system in a cerebellar-specific manner. Moreover, in Purkinje cells, CYP2J6 showed an elevated expression level compared to that of astrocytes, granule cells, and microglia. In this regard, we found that one CYP metabolite, 14,15-EET, acts as a key mediator in ameliorating cholesterol accumulation. In confirming this hypothesis, 14,15-EET treatment reduced the accumulation of cholesterol in human NPC1 patient-derived fibroblasts in vitro by suppressing cholesterol synthesis and ameliorating the impaired autophagic flux. We show that the reduced activity within the CYP system in the cerebellum could cause the neurological symptoms of NPC1 patients, as 14,15-EET treatment significantly rescued cholesterol accumulation and impaired autophagy. We also provide evidence that the intranasal administration of hUCB-MSCs is a highly promising alternative to traumatic surgical transplantation for NPC1 patients.

• Journal of Biomedical and Translational Research
• /
• 제19권4호
• /
• pp.116-124
• /
• 2018

Korean Native Pig (KNP) has a uniform black coat color, excellent meat quality, white colored fat, solid fat structure and good marbling. However, its growth performance is low, while the western origin Yorkshire pig has high growth performance. To take advantage of the unique performance of the two pig breeds, we raised crossbreeds (KNP ${\times}$ Yorkshire to make use of the heterotic effect. We then analyzed the liver transcriptome as it plays an important role in fat metabolism. We sampled at two stages: 10 weeks and at 26 weeks. The stages were chosen to correspond to the change in feeding system. A total of 16 pigs (8 from each stage) were sampled and RNA sequencing was performed. The reads were mapped to the reference genome and differential expression analysis was performed with edgeR package. A total of 324 genes were found to be significantly differentially expressed (${\left|log2FC\right|}$ > 1 & q < 0.01), out of which 180 genes were up-regulated and 144 genes were down-regulated. Principal Component Analysis (PCA) showed that the samples clustered according to stages. Functional annotation of significant DEGs (differentially expressed genes) showed that GO terms such as DNA replication, cell division, protein phosphorylation, regulation of signal transduction by p53 class mediator, ribosome, focal adhesion, DNA helicase activity, protein kinase activity etc. were enriched. KEGG pathway analysis showed that the DEGs functioned in cell cycle, Ras signaling pathway, p53 signaling pathway, MAPK signaling pathway etc. Twenty-nine transcripts were also part of the DEGs, these were predominantly Cys2His2-like fold group (C2H2) family of zinc fingers. A protein-protein interaction (PPI) network analysis showed that there were three highly interconnected clusters, suggesting an enrichment of genes with similar biological function. This study presents the first report of liver tissue specific gene regulation in a cross-bred Korean pig.

• 생명과학회지
• /
• 제31권10호
• /
• pp.954-959
• /
• 2021

세포 사멸은 항상성을 유지하기 위해 세포군을 조절하는 중요한 메커니즘이며 시스테인 단백질분해효소 중 하나인 카스파제는 세포 사멸 경로의 중요한 중재자이다. Caspase-8은 세포외 자극에 의해 시작되는 외인성 세포자멸 경로의 개시자 카스파제이다. Caspase-8에는 보존된 도메인인 N-말단의 두개의 죽음 이펙터 도메인(DED)과 C-말단의 2개의 촉매 도메인을 가지며, 이는 이러한 외인성 세포자멸 경로에 중요하게 작용한다. 외인성 세포멸사 경로에서, TNF 슈퍼패밀리인 죽음 수용체는 세포 외부로부터의 죽음 수용체 특이적 리간드의 결합에 의해 활성화된다. 활성화된 죽음 수용체가 어댑터 단백질인 Fas-associated death domain 단백질(FADD)을 모집한 후, 죽음 수용체와 FADD의 죽음 도메인(DD)이 서로 결합하고 죽음 수용체와 결합한 FADD가 caspase-8의 전구체 형태인 procaspase-8을 모집한다. FADD와 procaspase-8의 죽음 이펙터 도메인은 서로 결합하고 FADD에 결합된 procaspase-8은 prodomain의 절단에 의해 활성화된다. 이 죽음 수용체-FADD-caspase-8 복합체는 세포사멸 유도 신호복합체(DISC)라고 한다. 세포 FLICE 억제 단백질(c-FLIPs)은 세포사멸을 억제하는 역할과 촉진하는 역할을 모두 수행하여 caspase-8의 활성화를 조절하고 caspase-8 활성화는 caspase-3와 같은 작동자 카스파제를 활성화를 시킨다. 마지막으로 활성화된 작동자 카스파제는 DNA 분해, 핵 응축, 세포막 수포 및 카스파제 기질의 단백질 분해에 작용하여 세포사멸을 완료한다.