• 제목/요약/키워드: N-terminal analysis

검색결과 483건 처리시간 0.027초

복숭아순나방 성충 복부에서 발현하는 불포화효소의 RNA 간섭과 교미교란 (Mating Disruption of Grapholita molesta by RNA Interference of a Fatty Acid Desaturase Expressed in Adult Abdomen)

  • 김규순;정충렬;양창열;권기면;김용균
    • 한국응용곤충학회지
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    • 제56권1호
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    • pp.61-67
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    • 2017
  • 복숭아순나방(Grapholita molesta)은 두 가지 주요 성페로몬 성분(Z-8-dodecenyl acetate and E-8-dodecenyl acetate)을 갖고 있다. 이 성페로몬 성분의 생합성 과정 분석은 포화지방산의 10번 탄소에 이중결합을 합성하는 불포화효소($10{\Delta}$ DES)가 종 특이적 광학이성체 형성에 필수적이라고 제시하였다. 그러나 이 효소의 분자적 특징에 대해서 분석되지 않았다. 본 연구는 복숭아순나방 성페로몬 샘의 전사체에서 $10{\Delta}$ DES로 추정된 불포화효소(Gm-comp1575)의 단백질 기능 영역을 분석하였다. Gm-comp1575 유전자는 370개의 아미노산 서열 정보를 암호하고 있으며 분자량은 약 43.2 kDa 그리고 등전위점(pI)은 8.77로 추정되었다. 이 불포화효소는 4개의 막투과영역을 지니고 있으며, 6개의 탄수화물 결합 위치가 아미노 말단과 세포내 영역에서 갖는 것으로 추정되었다. 분자계통분석은 Gm-comp1575가 다른 종에서 알려진 $10{\Delta}$ DES와 유사성이 높은 것으로 밝혀졌다. Gm-comp1575 전사체는 암컷 성페로몬 샘 및 다른 복부 조직에서 발현되었다. 이 유전자 발현에 대한 RNA 간섭 처리는 처녀 암컷으로 하여금 사과원에서 수컷을 유인하는 능력을 크게 감소시켰다. 이러한 결과는 Gm-comp1575가 복숭아순나방의 성페로몬 생합성과 관련이 있는 유전자라고 제시하고 있다.

Biosynthesis of Isoprenoids: Characterization of a Functionally Active Recombinant 2-C-methyl-D-erythritol 4-phosphate Cytidyltransferase (IspD) from Mycobacterium tuberculosis H37Rv

  • Shi, Wenjun;Feng, Jianfang;Zhang, Min;Lai, Xuhui;Xu, Shengfeng;Zhang, Xuelian;Wang, Honghai
    • BMB Reports
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    • 제40권6호
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    • pp.911-920
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    • 2007
  • Tuberculosis, caused by Mycobacterium tuberculosis, continues to be one of the leading infectious diseases to humans. It is urgent to discover novel drug targets for the development of antitubercular agents. The 2-C-methyl-Derythritol-4-phosphate (MEP) pathway for isoprenoid biosynthesis has been considered as an attractive target for the discovery of novel antibiotics for its essentiality in bacteria and absence in mammals. MEP cytidyltransferase (IspD), the third-step enzyme of the pathway, catalyzes MEP and CTP to form 4-diphosphocytidyl-2-C-methylerythritol (CDP-ME) and PPi. In the work, ispD gene from M. tuberculosis H37Rv (MtIspD) was cloned and expressed. With N-terminal fusion of a histidine-tagged sequence, MtIspD could be purified to homogeneity by one-step nickel affinity chromatography. MtIspD exists as a homodimer with an apparent molecular mass of 52 kDa. Enzyme property analysis revealed that MtIspD has high specificity for pyrimidine bases and narrow divalent cation requirements, with maximal activity found in the presence of CTP and $Mg^{2+}$. The turnover number of MtIspD is $3.4 s^{-1}$. The Km for MEP and CTP are 43 and $92{\mu}M$, respectively. Furthermore, MtIspD shows thermal instable above $50^{\circ}C$. Circular dichroism spectra revealed that the alteration of tertiary conformation is closely related with sharp loss of enzyme activity at higher temperature. This study is expected to help better understand the features of IspD and provide useful information for the development of novel antibiotics to treat M. tuberculosis.

Photoaging protective effects of BIOGF1K, a compound-K-rich fraction prepared from Panax ginseng

  • Hong, Yo Han;Kim, Donghyun;Nam, Gibaeg;Yoo, Sulgi;Han, Sang Yun;Jeong, Seong-Gu;Kim, Eunji;Jeong, Deok;Yoon, Keejung;Kim, Sunggyu;Park, Junseong;Cho, Jae Youl
    • Journal of Ginseng Research
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    • 제42권1호
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    • pp.81-89
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    • 2018
  • Background: BIOGF1K, a compound-K-rich fraction, has been shown to display anti-inflammatory activity. Although Panax ginseng is widely used for the prevention of photoaging events induced by UVB irradiation, the effect of BIOGF1K on photoaging has not yet been examined. In this study, we investigated the effects of BIOGF1K on UVB-induced photoaging events. Methods: We analyzed the ability of BIOGF1K to prevent UVB-induced apoptosis, enhance matrix metalloproteinase (MMP) expression, upregulate anti-inflammatory activity, reduce sirtuin 1 expression, and melanin production using reverse transcription-polymerase chain reaction, melanin content assay, tyrosinase assay, and flow cytometry. We also evaluated the effects of BIOGF1K on the activator protein-1 signaling pathway, which plays an important role in photoaging, by immunoblot analysis and luciferase reporter gene assays. Results: Treatment of UVB-irradiated NIH3T3 fibroblasts with BIOGF1K prevented UVB-induced cell death, inhibited apoptosis, suppressed morphological changes, reduced melanin secretion, restored the levels of type I procollagen and sirtuin 1, and prevented mRNA upregulation of MMP-1, MMP-2, and cyclo-oxygenase-2; these effects all occurred in a dose-dependent manner. In addition, BIOGF1K markedly reduced activator-protein-1-mediated luciferase activity and decreased the activity of mitogen-activated protein kinases (extracellular response kinase, p38, and C-Jun N-terminal kinase). Conclusion: Our results strongly suggest that BIOGF1K has anti-photoaging activity and that BIOGF1K could be used in anti-aging cosmeceutical preparations.

임종돌봄에 대한 교육의 필요성: 2개 대학병원의 인턴을 대상으로 예비 분석 (The Necessity for End-of-Life Care Education: A Preliminary Analysis with Interns at Two University Hospitals)

  • 김도연;김경지;신성준;권복규;남은미;허대석;이순남
    • Journal of Hospice and Palliative Care
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    • 제20권2호
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    • pp.111-121
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    • 2017
  • 목적: 본 연구는 2개 대학병원의 인턴을 대상으로 임종돌봄에 대한 교육 현황을 파악하고자 하였다. 방법: 2개 대학병원의 64명의 인턴들에게 구조화된 설문지를 배포하여 임종돌봄 교과과정의 강의습득 경험과 임상견학 경험 여부를 확인하였다. 또한 임종돌봄 관련 교육이나 진료에 대한 태도를 평가할 수 있는 7개의 자가평가질문을 파악하였다. 결과: 임종돌봄 교과과정 강의 습득과 임상실습 견학 개수의 평균은 각각 5개(총 영역 개수: 9)와 2개(총 영역 개수: 7)였다. 가장 많이 교육받은 항목은 나쁜 소식 전하기(96.9%)였고 임상실습 동안 가장 많이 견학한 항목은 신체증상 조절(56.5%)였다. 그러나 사전연명의료의향서를 포함한 의사소통 영역에 대한 경험은 20% 미만이었다. 임종돌봄 단독 교과목 교육군은 그렇지 않은 군과 비교하여 임종돌봄 교과과정에 대한 만족도, 관심도, 준비됨의 7가지 항목에서 모두 유의하게 긍정적인 답변을 하였다. 결론: 본 연구는 임종돌봄이 필요한 환자를 진료해야 하는 인턴들이 임종돌봄에 대한 교과과정 습득과 임상경험이 매우 부족함을 관찰하였다. 추후 국내 임종돌봄 교육에 대한 의과대학교육의 전체적인 상황을 파악하여 관련된 적절한 지식과 기술을 연마하는 교육체계를 세울 필요가 있다.

실시간 Igr 검출을 위한 센서 모듈의 제작 및 신뢰성 평가에 관한 연구 (A Study on the Fabrication of the Sensor Module for the Detection of Resistive Leakage Current (Igr) in Real Time and Its Reliability Evaluation)

  • 이병설;최충석
    • 한국안전학회지
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    • 제33권1호
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    • pp.28-34
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    • 2018
  • The purpose of this study is to fabricate a sensor module to detect the resistive leakage current (Igr) in real time that occurs to low voltage electric lines and to verify its reliability. In the case of the developed sensor module, wires are inserted into the zero current transformer (ZCT) and current transformer (CT) in advance and then the branch line is connected to the circuit breaker. The measurement result of the resistance of the distribution panel equipped with the developed sensor module shows that the resistance is $0.151m{\Omega}$ between the R and R phases, $0.169m{\Omega}$ between the S and S phases, and $0.178m{\Omega}$ between the T and T phases, respectively. The insulation resistance measured at AC 500 V and 1,000 V is $0.08m{\Omega}$ between the R, S, T and N phases, respectively. Then, the insulation resistance measured at DC 500 V is $83.3G{\Omega}$ between the R, S, T and G terminal, respectively. In addition, the applied withstanding voltage is AC 220 V/380 V/440 V and it was found that characteristics between all phases are good. This study measured the standby power by installing the developed sensor module at the rear of the MCCB and switching the circuit breaker on sequentially. The standby power is 1.350 W when one circuit breaker is turned on, 1.690 W when 2 circuit breakers are turned on, and 4.371 W when 10 circuit breakers are turned on. This study also verified the reliability of the standby power of the distribution panel equipped with the developed sensor module using the Minitab Program (Minitab PGM). Since the analysis shows the statistical average of 1.34627 in the reliable range of normal distribution, standard deviation of 0.001874, AD of 0.554, and P value of 0.140, it is found that the distribution panel equipped with the developed sensor module has high reliability.

스트레스-유도 열충격단백질 27(Heat Shock Protein 27)의 활성과 물리치료의 상관성 (The Activation of Stress-induced Heat Shock Protein 27 and the Relationship of Physical Therapy)

  • 김미선;이성호;김일현;황병용;김중환
    • The Journal of Korean Physical Therapy
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    • 제20권1호
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    • pp.57-65
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    • 2008
  • Purpose: Heat shock proteins (HSPs) are a group of proteins that are activated when cells are exposed to a variety of environmental stresses, such as infection, inflammation, exposure to toxins, starvation, hypoxia, brain injury, or water deprivation. The activation of HSPs by environmental stress plays a key role in signal transduction, including cytoprotection, molecular chaperone, anti-apoptotic effect, and anti-aging effects. However, the precise mechanism for the action of small HSPs, such as HSP27 and mitogen-activated protein kinases (MAPKs: extracellular-regulated protein kinase 1/2 (ERK1/2), p38MAPK, stress-activated protein kinase/c-Jun N-terminal kinase (SAPK/JNK), is not completely understood, particularly in application of cell stimulators including platelet-derived growth factor (PDGF), angiotensin II (AngII), tumor necrosis factor $\alpha$ (TNF$\alpha$), and $H_2O_2$. This study examined the relationship between stimulators-induced enzymatic activity of HSP27 and MAPKs from rat smooth and skeletal muscles. Methods: 2-dimensional electrophoresis (2DE) and matrix assisted laser desorption ionizationtime-of-flight/time-of-flight (MALDI-TOF/TOF) analysis were used to identify HSP27 from the intact vascular smooth and skeletal muscles. Three isoforms of HSP27 were detected on silver-stained gels of the whole protein extracts from the rat aortic smooth and skeletal muscle strips. Results: The expression of PDGF, AngII, TNF$\alpha$, and $H_2O_2$-induced activation of HSP27, p38MAPK, ERK1/2, and SAPK/JNK was higher in the smooth muscle cells than the control. SB203580 (30${\mu}$M), a p38MAPK inhibitor, increased the level of HSP27 phosphorylation induced by stimulators in smooth muscle cells. Furthermore, the age-related and starvation-induced activation of HSP27 was higher in skeletal muscle cells (L6 myoblast cell lines) and muscle strips than the control. Conclusion: These results suggest, in part, that the activity of HSP27 and MAPKs affect stressors, such as PDGF, AngII, TNF$\alpha$, $H_2O_2$, and starvation in rat smooth and skeletal muscles. However, more systemic research will be needed into physical therapy, including thermotherapy, electrotherapy, radiotherapy and others.

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해인탕이 뇌허혈 유발 모래쥐의 단기기억력 감퇴와 치상회 세포사멸에 미치는 효과 (Neuroprotective Effects of Haein-tang(Hairen-tang) on Decrease of Short-term Memory and Apoptosis in Dentate Gyrus of the Gerbils with Transient Global Ischemia)

  • 박정철;송윤경;임형호
    • 한방재활의학과학회지
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    • 제21권2호
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    • pp.1-13
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    • 2011
  • Objectives : We investigated the effect of Haein-tang(Hairen-tang) on short-term memory and apoptosis in dentate gyrus of the gerbils with transient global ischemia. Methods : For the induction of cerebral ischemia model in mice, common carotid arteries of gerbils were occluded with aneurysm clips for 5 min. One day after operation, Haein-tang(Hairen-tang) was administrated orally injected once a day for 15 consecutive days. Gerbils were randomly divided into four group(n=10 in each group): sham-operation group, ischemia-induction group, ischemia-induction and 50 mg/kg Haein-tang(Hairen-tang)-treated group, ischemia-induction and 100 mg/kg Haein-tang(Hairen-tang)-treated group, and ischemia-induction and 200 mg/kg Haein-tang(Hairen-tang)-treated group. The effect of Haein-tang(Hairen-tang) on memory function was investigated by using step-down avoidance task. Apoptosis was confirmed by terminal deoxynucleotidyl transferase-mediated dUTP nick end labeling(TUNEL) staining and immunohistochemistry for caspase-3. Western blot analysis for the expressions of Bax and Bcl-2 protein was also conducted. Results : 1. Haein-tang extract significantly enhanced short-term memory in step-down avoidance task and 100 mg/kg, 200 mg/kg Haein-tang-treated group. 2. Haein-tang extract significantly suppressed TUNEL-positive cells after transient global ischemia and 50 mg/kg, 100 mg/kg, 200 mg/kg Haein-tang-treated group. 3. Haein-tang extract significantly increased caspase-3 positive cells in the hippocampal dentate gyrus after transient global ischemia and 50 mg/kg, 100 mg/kg, 200 mg/kg Haein-tang-treated group. 4. Haein-tang extract significantly decreased Bax protein expressions in the hippocampus after transient global ischemia and 100 mg/kg, 200 mg/kg, Haein-tang-treated group. Haein-tang extract significantly increased Bcl-2 protein expressions in the hippocampal dentate gyrus after transient global ischemia and 50 mg/kg, 100 mg/kg, 200 mg/kg, Haein-tang-treated group. Haein-tang extract significantly decreased Ratio of Bax protein to Bcl-2 protein in the hippocampus after transient global ischemia and 100 mg/kg, 200 mg/kg Haein-tang-treated group. Conclusions : While Haein-tang(Hairen-tang) treatment improved short-term memory by suppressing on ischemia-induction apoptosis. In the present study, Haein-tang(Hairen-tang) shows protective effect on transient global ischemia.

Palmitic acid에 의한 간세포 사멸효과에 대한 p38 MAPK 및 JNK 관련성 (The Involvement of p38 MAPK and JNK Activation in Palmitic Acid-Induced Apoptosis in Rat Hepatocytes)

  • 배춘식;박수현
    • 생명과학회지
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    • 제19권8호
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    • pp.1119-1124
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    • 2009
  • 포화 지방산이 인슐린 저항성 및 지방간등의 다양한 질병의 발병에 관련된다고 보고되고 있다. 그러나 간세포에서 포화지방산이 세포 사멸에 대한 효과 및 이와 관련된 신호전달계에 대해서는 거의 알려져 있지 않고 있다. 본 연구에서는 대표적인 포화지방산인 palmitic acid 처리 시 랫트 간세포 사멸에 미치는 효과와 이들이 mitogen activated protein kinases (MAPKs)와 관련되는 지를 알아보았다. 본 실험에서 palmitic acid 처리 시 간세포 성장은 억제 되었고 lactate dehydrogenase (LDH) 활성은 증가하였다. 이러한 작용은 JNK 및 p38 MAPK 억제제에 의해서 선택적으로 차단되었다. 아울러 palmitic acid에 의한 Bcl-2 발현 억제, Bax 발현 증가 작용, GSH 함량 감소작용 및 산화성 스트레스 증가작용 역시 JNK 및 p38 MAPK 억제제에 의해서 선택적으로 차단되었다. 실제로 palmitic acid 처리시 JNK 및 p38 MAPK 활성은 증가하였다. 결론적으로 palmitic acid는 간세포에서 JNK 및 p38 MAPK 활성을 경유하여 산화성 스트레스 증가를 통하여 간세포 사멸을 유도하는 것으로 나타났다.

Gain of a New Exon by a Lineage-Specific Alu Element-Integration Event in the BCS1L Gene during Primate Evolution

  • Park, Sang-Je;Kim, Young-Hyun;Lee, Sang-Rae;Choe, Se-Hee;Kim, Myung-Jin;Kim, Sun-Uk;Kim, Ji-Su;Sim, Bo-Woong;Song, Bong-Seok;Jeong, Kang-Jin;Jin, Yeung-Bae;Lee, Youngjeon;Park, Young-Ho;Park, Young Il;Huh, Jae-Won;Chang, Kyu-Tae
    • Molecules and Cells
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    • 제38권11호
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    • pp.950-958
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    • 2015
  • BCS1L gene encodes mitochondrial protein and is a member of conserved AAA protein family. This gene is involved in the incorporation of Rieske FeS and Qcr10p into complex III of respiratory chain. In our previous study, AluYRa2-derived alternative transcript in rhesus monkey genome was identified. However, this transcript has not been reported in human genome. In present study, we conducted evolutionary analysis of AluYRa2-exonized transcript with various primate genomic DNAs and cDNAs from humans, rhesus monkeys, and crabeating monkeys. Remarkably, our results show that AluYRa2 element has only been integrated into genomes of Macaca species. This Macaca lineage-specific integration of AluYRa2 element led to exonization event in the first intron region of BCS1L gene by producing a conserved 3' splice site. Intriguingly, in rhesus and crabeating monkeys, more diverse transcript variants by alternative splicing (AS) events, including exon skipping and different 5' splice sites from humans, were identified. Alignment of amino acid sequences revealed that AluYRa2-exonized transcript has short N-terminal peptides. Therefore, AS events play a major role in the generation of various transcripts and proteins during primate evolution. In particular, lineage-specific integration of Alu elements and species-specific Alu-derived exonization events could be important sources of gene diversification in primates.

Association of a Methanol Extract of Rheum undulatum L. Mediated Cell Death in AGS Cells with an Intrinsic Apoptotic Pathway

  • Hong, Noo Ri;Park, Hyun Soo;Ahn, Tae Seok;Jung, Myeong Ho;Kim, Byung Joo
    • 대한약침학회지
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    • 제18권2호
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    • pp.26-32
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    • 2015
  • Objectives: Rheum undulatum L. has traditionally been used for the treatment of many diseases in Asia. However, its anti-proliferative activity in cancer has still not been studied. In the present study, we investigated the anti-cancer effects of methanol extract of Rheum undulatum L. (MERL) on human adenocarcinoma gastric cell lines (AGS). Methods: To investigate the anti-cancer effect of MERL on AGS cells, we treated the AGS cells with varying concentrations of MERL and performed 3-(4, 5-dimethylthiazol-2-yl)-2, 5-diphenyltetrazolium bromide (MTT) assays. Cell cycle analyses, measurements of the mitochondrial membrane potential (MMP), caspase activity assays and Western blots were conducted to determine whether AGS cell death occurred by apoptosis. Results: Treatment with MERL significantly inhibited growth of AGS cells in a concentration dependent manner. MERL treatment in AGS cells leaded to increased accumulation of apoptotic sub G1 phase cells in a concentration dependent manner. In control cultures, 5.38% of the cells were in the sub G1 phase. In MERL treated cells, however, this percentage was significantly increased (9.95% at $70{\mu}g/mL$, 15.94% at $140{\mu}g/mL$, 26.56% at $210{\mu}g/mL$ and 38.08% at $280{\mu}g/mL$). MERL treatment induced the decreased expression of pro-caspase-8 and -9 in a concentration dependent manner, whereas the expression of the active form of caspase-3 was increased. A subsequent Western blot analysis revealed increased cleaved levels of poly (ADP-ribose) polymerase (PARP) protein. Also, treatment with MERL increased the activities of caspase-3 and -9 compared with the control. MERL treatment increased the levels of the pro-apoptotic truncated Bid (tBid) and Bcl2 Antagonist X (Bax) proteins and decreased the levels of the anti-apoptotic B-cell lymphoma 2 (Bcl-2) protein, whose is the stabilization of mitochondria. However, inhibitions of p38, extracellular signal regulated kinases (ERKs) and C-Jun N-terminal kinases (JNK) by MERL treatment did not affect cell death. Conclusion: These results suggest that MERL mediated cell death is associated with an intrinsic apoptotic pathway in AGS cells.