• Title/Summary/Keyword: atpB

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Background $K^+$ channel currents in WEHI-231 cells, immature B lymphocytes

  • Nam, Joo-Hyun;Woo, Ji-Eun;Kim, Tae-Jin;Uhm, Dae-Yong;Kim, Sung-Joon
    • Proceedings of the Korean Biophysical Society Conference
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    • 2003.06a
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    • pp.39-39
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    • 2003
  • In our previous study, WEHI-231, an immature B cell line, showed intractable increase in [C $a^{2+}$]$_{c}$ after the B-cell receptor (BCR) ligation and treatment with 2-aminoethoxydiphenylborate (2-APB), which was never observed in Bal-17, a mature B cell line (Nam et al., 2003, FEBS Lett). In this study, a whole cell voltage clamp study revealed a specific expression of a novel type of $K^{+}$ current, namely voltage-independent background-type $K^{+}$ channels (IK-bg), in WEHI-231 cells. IK-bg was dramatically increase by the application of 2-APB (50 $\square$M), which induced severe hyperpolarization of WEHI-231 from -45 ㎷ to -90 ㎷, When dialyzed with $Mg^{2+}$ and ATP-free pipette solution, a spontaneous development of IK-bg and membrane hyperpolarization were observed. IK-bg was insensitive to classical $K^{+}$ channel blockers (TEA, glibenclamide, $Ba^{2+}$(1 mM)), whereas blocked by quinine and quinidine in a voltage-dependent manner ($IC_{50}$/=6~9 $\square$M at +60㎷). Phorbol myrstate, a PKC activator, decreased the amplitude of IK-bg. Extracellular acidification (pH 6.5) slightly inhibited IK-bg. Arachidonic acid, riluzole, or hyposmotic stress could not affect the IK-bg after the full development by the intracellular dialysis with Mg-ATP-free solution. In a cell-attached mode of single channel recording from WEHI231, we found two types of voltage-independent $K^{+}$ channels with unitary conductance of 300 pS and 120 pS, respectively. Both channels showed very short mean open times and their open probabilities were increase by the application of 2-APB. In Bal-17 cells, no such $K^{+}$ current was observed in 50 cells tested. In summary, WEHI-231 immature B cells express background $K^{+}$ channels. The pharmacological properties and the large unitary conductance suggest that novel types of two-pore domain $K^{+}$ channels (2-P-K channels) might be expressed in WEHI-231, which may provide an intriguing targets of signal transduction in the immature B lymphocytes.e B lymphocytes.

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Beneficial Effect of Coumestrol on Ultraviolet B-Induced Skin Photoaging through Mitochondrial Biogenesis (쿠메스트롤의 미토콘드리아 생합성 증가를 통한 피부 광노화 예방 효과)

  • Kim, Su-Kyung;Kim, Jeong-Kee;Seo, Dae-Bang;Lee, Sang-Jun
    • Journal of the Society of Cosmetic Scientists of Korea
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    • v.38 no.3
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    • pp.237-245
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    • 2012
  • Coumestrol is one of phytoalexins synthesized in response to environmental stress, and commonly found in natural foods such as alfalfa sprouts, clovers, and soybean. In the present study, we investigated the mechanism underlying protective effect of coumestrol against UVB-induced photoaging in human dermal fibroblasts. We found that pretreatment with coumestrol enhanced the UVB-suppressed mitochondrial biogenesis through regulation of Sirt1 expression and activity, and its downstream gene regulation such as PGC-$1{\alpha}$, NRF1, and TFAM. Moreover, the ATP and ROS production was restored to normal status and the formation of advanced glycation endproducts leading to skin photoaging in skin fibroblasts was blocked by coumestrol pretreatment before UVB irradiation. These findings indicate that coumestrol might potentially prevent skin photoaging induced by mitochondrial damage and glycated protein production in dermal fibroblasts.

Isolation and Characterization of the Colletotrichum acutatum ABC Transporter CaABC1

  • Kim, Suyoung;Park, Sook-Young;Kim, Hyejeong;Kim, Dongyoung;Lee, Seon-Woo;Kim, Heung Tae;Lee, Jong-Hwan;Choi, Woobong
    • The Plant Pathology Journal
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    • v.30 no.4
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    • pp.375-383
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    • 2014
  • Fungi tolerate exposure to various abiotic stresses, including cytotoxic compounds and fungicides, via their ATP-driven efflux pumps belonging to ATP-binding cassette (ABC) transporters. To clarify the molecular basis of interaction between the fungus and various abiotic stresses including fungicides, we constructed a cDNA library from germinated conidia of Colletotrichum acutatum, a major anthracnose pathogen of pepper (Capsicum annum L.). Over 1,000 cDNA clones were sequenced, of which single clone exhibited significant nucleotide sequence homology to ABC transporter genes. We isolated three fosmid clones containing the C. acutatum ABC1 (CaABC1) gene in full-length from genomic DNA library screening. The CaABC1 gene consists of 4,059 bp transcript, predicting a 1,353-aa protein. The gene contains the typical ABC signature and Walker A and B motifs. The 5'-flanking region contains a CAAT motif, a TATA box, and a Kozak region. Phylogenetic and structural analysis suggested that the CaABC1 is a typical ABC transporter gene highly conserved in various fungal species, as well as in Chromista, Metazoans, and Viridiplantae. We also found that CaABC1 was up-regulated during conidiation and a minimal medium condition. Moreover, CaABC1 was induced in iprobenfos, kresoxim-methyl, thiophanate-methyl, and hygromycin B. These results demonstrate that CaABC1 is necessary for conidiation, abiotic stress, and various fungicide resistances. These results will provide the basis for further study on the function of ABC transporter genes in C. acutatum.

Molecular Genetic Testing and Diagnosis of Wilson Disease (윌슨병의 진단과 분자유전학적 검사)

  • Seo, Jeong Kee
    • Pediatric Gastroenterology, Hepatology & Nutrition
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    • v.11 no.sup1
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    • pp.72-82
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    • 2008
  • Wilson disease (WD) is an autosomal recessive disorder of copper metabolism that results in accumulation of copper primarily in the liver, the brain and the cornea. Mutations in the WD gene, ATP7B cause failure of copper excretion from hepatocyte into bile and a defective synthesis of ceruloplasmin. More than 370 mutations are now recognized, scattering throughout the ATP7B gene. Since WD has protean clinical presentations, awareness of WD in clinical practice is important for the early diagnosis and prevention of accumulated copper toxicity. None of the laboratory parameters alone allows a definite diagnosis of WD. There are numerous pitfalls in the diagnosis of WD. Low serum ceruloplasmin concentrations, increased 24 hour urinary copper excretion, increased hepatic copper concentrations and the presence of Kayser-Fleischer rings in the cornea are major diagnostic points. A combination of any two of these 4 laboratory findings is strong support for a diagnosis of WD. Molecular methods are now being used to aid diagnosis. Molecular genetic testing has confirmed the diagnosis in individuals in whom the diagnosis is not clearly established biochemically and clinically. Siblings should be screened for WD once an index case has been diagnosed. Discrimination of heterozygotes from asymptomatic patients is essential to avoid inappropriate lifelong therapy for heterozygotes. Genetic testing, either by haplotype analysis or by mutation analysis, is the only reliable tool for differentiating heterozygote carriers from affected asymptomatic patients. Currently, genetic testing is of limited value in the primary diagnosis. However, genetic testing will soon play an essential role in diagnosing WD as rapid advancement of biomedical technology will allow more rapid, easier and less expensive mutation detection.

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Molecular phylogenetic relationship of the family Colchicaceae (Liliales)

  • Thi, Nguyen Pham Anh;Kim, Jung-Sung;Kim, Joo-Hwan
    • Proceedings of the Plant Resources Society of Korea Conference
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    • 2012.05a
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    • pp.19-19
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    • 2012
  • The Colchicaceae comprising 250 species and 15-19 genera of rhizomatous or cormous perennials, the moderate sized family in Liliales, distributes widely through the temperate and tropical areas of Africa, Asia and North America. The division of two subfamilies in Colchicaceae is still unclear because of different results in previous studies. Moreover, sister taxa of this family has not been determined. In genus level, it was uncertain that whether expand circumscription of three genera of Colchicum, Gloriosa, and Wurmbea which are include Androcymbium, Littonia and Onixotis, respectively, is reasonable or not. In this study, three coding genes of atpB, matK and rbcL were analyzed to reconstruct phylogenetic relationship of Colchicaceae and both of maximum parsimony (MP) and Bayesian analysis were conducted. Among three genes, matK region was most variable and provided more parsimony-informative sites, whereas the atpB and rbcL regions were similar in the variation and number of informative characters. Monophyly of Colchicaceae was strongly supported and it was divided into two subfamilies (Wurmbeoideae and Uvulariodeae). Uvularia-Disporum clade, comprises the subfamily Uvularioideae, is a sister of the rest Colchicaceae and subsequently differentiated Burchardia was a sister within subfamily Wurmbeoideae. Burchadia was used to be supposed to be a sister of the family in the previous studies. It was clear the monophyly and phylogenetic relationship among six tribes sensu Vinnersten and Manning (2007) within the family. In addition, the expanded circumscription of three genera was also strongly supported; Colchicum-Androcymbium (BP99), Wurmbea-Onixotis (BP100), and Littonia-Gloriosa (BP100). Here, we propose a re-circumscription among taxa of Colchicaceae.

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Selective Interaction Between Chloroplast β-ATPase and TGB1L88 Retards Severe Symptoms Caused by Alternanthera mosaic virus Infection

  • Seo, Eun-Young;Nam, Jiryun;Kim, Hyun-Seung;Park, Young-Hwan;Hong, Seok Myeong;Lakshman, Dilip;Bae, Hanhong;Hammond, John;Lim, Hyoun-Sub
    • The Plant Pathology Journal
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    • v.30 no.1
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    • pp.58-67
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    • 2014
  • The multifunctional triple gene block protein 1 (TGB1) of the Potexvirus Alternanthera mosaic virus (AltMV) has been reported to have silencing suppressor, cell-to-cell movement, and helicase functions. Yeast two hybrid screening using an Arabidopsis thaliana cDNA library with TGB1 as bait, and co-purification with TGB1 inclusion bodies identified several host proteins which interact with AltMV TGB1. Host protein interactions with TGB1 were confirmed by biomolecular fluorescence complementation, which showed positive TGB1 interaction with mitochondrial ATP synthase delta' chain subunit (ATP synthase delta'), light harvesting chlorophyll-protein complex I subunit A4 (LHCA4), chlorophyll a/b binding protein 1 (LHB1B2), chloroplast-localized IscA-like protein (ATCPISCA), and chloroplast ${\beta}$-ATPase. However, chloroplast ${\beta}$-ATPase interacts only with $TGB1_{L88}$, and not with weak silencing suppressor $TGB1_{L88}$. This selective interaction indicates that chloroplast ${\beta}$-ATPase is not required for AltMV movement and replication; however, TRV silencing of chloroplast ${\beta}$-ATPase in Nicotiana benthamiana induced severe tissue necrosis when plants were infected by AltMV $TGB1_{L88}$ but not AltMV $TGB1_{L88}$, suggesting that ${\beta}$-ATPase selectively responded to $TGB1_{L88}$ to induce defense responses.

The Effects of Saengkankunbi-tang and Its Composition on Free Fatty Acid-Induced Lipotoxicity in HepG2 Cell (HepG2 cell에서 유리지방산 유발 지방독성에 대한 생간건비탕(生肝健脾湯)과 일부 조성 한약물의 효과)

  • Hong, Sung-In
    • The Journal of Internal Korean Medicine
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    • v.34 no.1
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    • pp.14-30
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    • 2013
  • Objectives : The aim of this study was to investigate whether the effects of extract from Saengkankunbi-tang and its composition that Artimisiae capillaris Herba, Crataegi Fructus, Alismatis Rhizoma, Hoelen and Raphani Semen could protect HepG2 cells from palmitic acid-induced lipotoxicity through lysosomal and mitochondrial pathways in an in vitro model. Methods : To examine the effects of the extracts from Saengkankunbi-tang and its composition that Artimisiae capillaris Herba, Crataegi Fructus, Alismatis Rhizoma, Hoelen and Raphani Semen on palmitic acid-induced lipotoxicity in HepG2 cells, we measured the contents of cell viability, cytotoxicity. Then to investigate the effects of the extract from Saengkankunbi-tang, Artimisiae capillaris Herba and Raphani Semen, we measured that triglyceride, reactive oxygen species, ATP levels, glutathione levels, cytochrome c and cathepsin B. Results : The extracts from Saengkankunbi-tang and its composition had a cell-protective function. The extracts from Saengkankunbi-tang, Artimisiae capillaris Herba and Raphani Semen controlled triglyceride over-accumulation in cells and reduced overproduction of reactive oxygen species. The extracts from Saengkankunbi-tang and Raphani Semen increased ATP and glutathione levels which had been decreased by lipotoxicity. The extracts from Saengkankunbi-tang, Artimisiae capillaris Herba and Raphani Semen reduced leakage of cytochrome c and the extracts from Saengkankunbi-tang and Raphani Semen reduced leakage of cathepsin B in lipotoxicity. Conclusions : These results show that the extracts from Saengkankunbi-tang and its composition that Artimisiae capillaris Herba and Raphani Semen have cell protective effects on palmitic acid-induced lipotoxicity through lysosomal and mitochondrial pathways.

Effect Of Nelumbinis Semen On The Recovery Of The Cardiac Muscle Activity by Proteome Analysis (연자육(蓮子肉)의 심근 경색 모델에 대한 Proteom 분석)

  • Ahn, Chang-Joon;Lee, Gi-Hyun;Kim, Yang-Seok;Hong, Moo-Chang;Bae, Hyun-Su;Kim, Jong-Hoon;Shin, Min-Kyu
    • Journal of Physiology & Pathology in Korean Medicine
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    • v.24 no.6
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    • pp.962-969
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    • 2010
  • The purpose of this investigation was to confirm the effect of Nelumbinis Semen on the recovery of the cardiac muscle activity. We studied the effect of Nelumbinis Semen on the recovery of ischemic SD rat hearts perfused with Nelumbinis Semen, using a model of ex-vivo perfusion (Non-working Langendorff perfusion system) and working heart perfusion system at the same time. To explore the effect of Nelumbinis Semen at the level of proteome, two-dimensional electrophoresis and MALDI-TOF analysis were performed. We found out that the proteins increased after perfusion of Nelumbinis Semen are Mitochondrial aconitase, ATP synthase alpha chain, Lactate dehydrogenase B, Creatine kinase, Glyceraldehyde 3-phosphate dehydrogenase, Alpha B-crystallin, Myosin and Heart fatty acid binding protein. Almost, all of them are concerned with ATP production in the cardiac muscle with glucose metabolism.

A taxonomic note of the genus Lilium section Martagon in Korea (한국산 백합속(Lilium) 내 말나리절의 분류학적 검토)

  • Song, Ji-Hee;Lee, Woong-Bin
    • Korean Journal of Plant Taxonomy
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    • v.47 no.1
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    • pp.46-50
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    • 2017
  • The genus Lilium comprises about 100 species and is a major element of the family Liliaceae in temperate region of North Hemisphere. In the light of recent studies on the genus, we tried to examine the taxonomy of three Korean species (L. hansonii, L. tsingtauense, and L. distichum) belonging to section Martagon. The four available chloroplast regions (rbcL, matK, ndhF, and atpB) in Lilium species were selected and analyzed for a maximum likelihood phylogeny. As a results, three Korean species of section Martagon formed a monophyletic group. However, they nested within paraphyletic section Sinomartagon. Further, the discussions on taxonomic affinities of Korean species have made based on comparison between topology of phylogenetic tree and morphology.

Phylogenetic Relationship between Two Taxa of Opuntia in Jeju Island (제주도 부채(손바닥)선인장속(Opuntia) 두 분류군의 계통발생학적 유연관계)

  • Yang, Young-Soo;Oh, Hong-Shik
    • Korean Journal of Plant Resources
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    • v.34 no.5
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    • pp.451-461
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    • 2021
  • In Jeju Island, there are two different types of Opuntia cactus in cladodae, fruit, flower, flowering period and spine. There is no comparative study of these two taxa yet, so it is often seen as the same species. The purpose of this study was to investigate the phylogenetic relationship between the Taxa of the Opuntia monacantha Haw. and Opuntia stricta Haw. in Jeju Island. The results indicate that the two taxa were completely different from each other on the analysis of nrITS, matK and atpB-rbcL gene as well as the morphological comparison. Also, O. monacantha among the two taxa is included in the Elatae series, so South America is the origin, and O. stricta is included in the Scheerianae series, which confirms that North America is the origin. The name of the O. monacantha in Jeju Island was named 'Jeju-Baick-nyun-cho' considering geographical location conditions and symbolic characteristics.