• Title/Summary/Keyword: Extracellular

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Pattern-Recognition Receptor Signaling Initiated From Extracellular, Membrane, and Cytoplasmic Space

  • Lee, Myeong Sup;Kim, Young-Joon
    • Molecules and Cells
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    • 제23권1호
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    • pp.1-10
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    • 2007
  • Invading pathogens are recognized by diverse germline-encoded pattern-recognition receptors (PRRs) which are distributed in three different cellular compartments: extracellular, membrane, and cytoplasmic. In mammals, the major extracellular PRRs such as complements may first encounter the invading pathogens and opsonize them for clearance by phagocytosis which is mediated by membrane-associated phagocytic receptors including complement receptors. The major membrane-associated PRRs, Toll-like receptors, recognize diverse pathogens and generate inflammatory signals to coordinate innate immune responses and shape adaptive immune responses. Furthemore, certain membrane-associated PRRs such as Dectin-1 can mediate phagocytosis and also induce inflammatory response. When these more forefront detection systems are avoided by the pathogens, cytoplasmic PRRs may play major roles. Cytoplasmic caspase-recruiting domain (CARD) helicases such as retinoic acid-inducible protein I (RIG-I)/melanoma differentiation-associated gene 5 (MDA5), mediate antiviral immunity by inducing the production of type I interferons. Certain members of nucleotide-binding oligomerization domain (NOD)-like receptors such as NALP3 present in the cytosol form inflammasomes to induce inflammatory responses upon ligand recognition. Thus, diverse families of PRRs coordinately mediate immune responses against diverse types of pathogens.

세포외 기질 단백질이 생쥐 단위발생란의 체외 발달에 미치는 영향 (Effect of Extracellular Matrix Proteins on the In Vitro Development of Parthenogenetic Mouse Eggs)

  • 곽대오;김선구;김영수;박충생
    • 한국수정란이식학회지
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    • 제8권2호
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    • pp.83-90
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    • 1993
  • To investigate the effect of extracellular matrix proteins on the in vitro development of ethanol-induced parthenogenetic eggs of ICR strain mice, those were cultured in vitro in fibronectin, gelatin, or collagen precoated culture dishes containing 1.5 ml of NaH-C03$_3$-BMOC-3 medium at 37$^{\circ}C$ for 96 hrs. under the atmosphere of 5% $CO_2$ and 95% air. Fibronectin, gelatin, or collagen significantly(P$\pm$1.4, 45.4i1.4, and 44.8$\pm$O.9, respectively. And the diameter of those eggs ranged 104.6$\pm$1.9, 102.8$\pm$2.3, and 103.4$\pm$O.8 $\mu$m, respectively.

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Isolation of Chitin-utilizing Bacterium and Production of Its Extracellular Chitinase

  • Woo, Cheol-Joo;Yun, Un-Jung;Park, Heul-Doung
    • Journal of Microbiology and Biotechnology
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    • 제6권6호
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    • pp.439-444
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    • 1996
  • A bacterial strain, designated as WY22, producing extracellular chitinase was isolated from the soil around the Youngduck area, after enrichment culture in a medium containing $1{\%}$ (w/v) wet colloidal chitin as a sole carbon source. The isolate was identified as a strain of Bacillus sp. based on its morphological and physiological characteristics. It was observed that Bacillus sp. WY22 could inhibit the growth of Fusarium oxysporum with hyphal extention-inhibition assay on potato dextrose agar plate supplemented with $1{\%}$ collidal chitin. Optimum culture conditions of Bacillus sp. WY22 were examined for chitinase production in a chitin medium. High level production of chitinase was observed not only in the chitin medium but in a medium supplemented with $1{\%}$ N-glucosamine or lactose instead of chitin. The optimum concentrations of colloidal chitin and yeast extract were 3.0 and $0.5{\%}$, and the optimum culture conditions for initial pH of medium and temperature were 7.0 and $30^{\circ}C$, respectively, for the production of chitinase.

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Bacillus subtilis의 단백질 분비기구 SecY의 유전자 수준의 조절이 단백질 분비에 미치는 영향

  • 김상숙;김순옥;서주원
    • 한국미생물·생명공학회지
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    • 제24권4호
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    • pp.408-414
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    • 1996
  • The SecY is a central component of the protein export machinery that mediate the translocation of secretory proteins across the plasma membrane, and has been known to be rate-limiting factor of secretion in Escherichia coli. In order to study the extracellular protein secretion in Gram-positive microorganism, we have, constructed strains harboring more than one copy of the gene for SecY. Firstly, the gene, for B. subtilis SecY and its promoter region was subcloned into pDH32 and the chimeric vector was inserted into amyE locus by homologous recombination. Secondly, low copy number vector, pCED6, was also used for subcloning the secY gene and for constructing a strain which harbors several copies of secY. The KH1 cell which harbor two copies of secY on the chromosome excreted more extracellular proteins than the wild type PB2. Moreover, the KH2 cells which harbor several copies of secY in pCED6 vector excreted more extracellular proteins than the KH1 cells. Here, we found that the capacity of protein secretion is partly controlled by the number of secY and it is suggested that SecY has also an important role in protein secretion in B. subtilis, a gram positive microorganism, as like in E. coli. This will promote the use of B. subtilis as a host for the expression of useful foreign gene and excretion of precious proteins.

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세포막 활동전압에서 음양(陰陽)의 상호관계 (Interrelation of Yin and Yang in Action Potential of Cell Membrane)

  • 박선영;김호현
    • 동의생리병리학회지
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    • 제27권5호
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    • pp.563-569
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    • 2013
  • This study was undertaken to apply the yin-yang theory in action potential. In order to apply the yin-yang theory in action potential, nature of yin and yang, interrelation of yin and yang and action potential in cell were reviewed. According to the yin-yang theory, inner cellular space corresponds to yin, but outer cellular space corresponds to yang. If we classify ions in intracellular fluid or extracellular fluid by nature of yin and yang, potassium(K+) corresponds to yang within yin(陰中之陽), protein(Pr-) corresponds to yin within yin(陰中之陰) in intracellular fluid, and sodium(Na+) corresponds to yang within yang(陽中之陽), chloride(Cl-) corresponds to yin within yang(陽中之陰) in extracellular fluid. Double donnan equilibrium and equilibrium potential were caused by intracellular anion(Pr-) and extracellular cation(Na+) are related with mutual rooting of yin and yang(陰陽互根) and opposition of yin and yang(陰陽對立). The influx and efflux of ion through cell membrane means waxing and waning of yin and yang(陰陽消長), the change of membrane potential means yin-yang conversion(陰陽轉化) during action potential.

세포외 Adenosine Deaminase를 생산하는 방선균의 분리 (Isolation of Actinomycetes Producing Extracellular Adenosin Deaminase)

  • 전홍기;김태숙
    • 미생물학회지
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    • 제28권1호
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    • pp.83-89
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    • 1990
  • 토양으로부터 세포의 adcnosinedeaminase를 생산하는 두종의 방선균 J-845S주와 J-326TK 주름 분리하여 각각 Streptomycs sp. J-845S와 Nocardioides sp. J326TK로 동정하였다. Streptomyces sp. J-845S는 그람염색 양성, 비항산성균으로 기균사의 흰색계열의 간상형의 비운동성의 포자를 형성하였으며, 포자의 표면은 평할하였고 나선상의 포자연쇄를 형성하였다. 균사체의 분자는 양호하였다. 세포벽 구성성분을 분석한 결과 LL-diaminopimelic acid를 함유하는 cell wall type I 이었으며, phospholipid type II였다. Nocardioides sp. J-326TK는 그람염색반응 양성, 항산성염색반응 음성이었으며, 균사체는 불규칙한 간상 또는 구사으이 절편으로 분절되었다. 기균사의 분자는 그다지 인정되지 않았으며, 포자형태의 긴 elements의 표면은 평할하였다. 세포벽 구성성분을 검토한 결과 cell wall type I과 phospholipid type I으로나타났다.

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Phenotype of Hepartocyte Spheroids in Synthetic Thermo-reversible Extracellular Matrix

  • Park, Keun--Hong;Park, Ju-Young;Bae, You-Han
    • Biotechnology and Bioprocess Engineering:BBE
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    • 제6권4호
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    • pp.264-268
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    • 2001
  • Aggregates of specific cells are often regarded as better from in artificial organs and mammalian cell bioreactors in terms of cell-specific functionality. In this study, the morphology and liver specific functions of freshly harvested primary rat hepatocytes, which were cultivated as spheroids and entrapped in a synthetic thermo-reversible extracellular matrix, were examined and compared to a control (hepatocytes in single cell form). A copolymer of N-isopropylacrylamide(98 mole % in feed) and acrylic acid (poly (NiPAAm-co-AAC)), a thermo- reversible copolymer gel ma- trix, was used to entrap hepatocytes either in spheroids or single cells. During a 7-day culture pe-riod, the spheroids maintained higher viability and produced albumin and urea at a relatively con-stant rate, while, the single cell culture showed a slight increase in cell numbers and a reduction in albumin secretion Hepatocytes cultrured as spheroids present a potentially useful three-dimensional cell culture system for application in bioartificial liver device.

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Cloning and Expression of the Extracellular $\beta$-lactamase gene from streptomyces sp. SMF13 in streptomyces lividans

  • Rak, Choi-Sang;Lee, Kye-Joon
    • 미생물학회지
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    • 제30권3호
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    • pp.149-153
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    • 1992
  • Cloning of the gene encoding extracellular .betha.-lactamase from Streptomyces sp. SMF13 in a plasmid pIJ702 and expression of the gene in Streptomyces invidans were carried out. Optimal conditions for the formation of protoplasts of S.lividans and the regeneration of the protoplasts were evaluated. Streptomyces sp. SMF-13 was selected as a donor strain of .betha.-lactamase gene and totla DNA of the strain was partially digested with Sau3A I. DNA fragments ranged from 4kb to 10 kb were ligated to pIJ702 AT Bgl II site and then the ligated DNAs were transformed to the protoplasts of S, livivans. The transformation efficiency was $2 *10^{3}$ .$\mu$g DNA for the ligated DNA mixture. One colony among a thousand colonies regenerated showed extracellular .betha.-lactamase and the size of the inserted DNA fragment was estimated to be 3.94 kb. The .betha.-lactamase activity in the culture broth of the recombinant strain was maximum at 3 days culture to be 1.0 unit/ml.

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Mapping of the Interaction Domain of DNA Topoisomerase $II{\alpha}$ and $II{\beta}$ with Extracellular Signal-Regulated Kinase 2

  • Park, Gye-Hwa;Bae, Young-Seuk
    • BMB Reports
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    • 제34권1호
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    • pp.85-89
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    • 2001
  • Both topoisomerase $II{\alpha}$ and $II{\beta}$ east as phosphoproteins in the cells. Recently it was reported that DNA topoisomerase $II{\alpha}$ associates with and is phosphorylated by the extracellular signal-regulated kinase 2 (ERK2). Also, ERK2 stimulates the activity of topoisomerase II by a phosphorylation-independent manner [Shapiro et al., (1999) Mol. Cell. Biol. 19, 3551-3560]. In this study, a yeast two-hybrid system was used to investigate the binding site between topoisomerase $II{\alpha}$ or $II{\beta}$ and ERK2. The two-hybrid test clearly showed that topoisomerase $II{\beta}$ residues 1099-1263, and topoisomerase $II{\alpha}$ residues 1078-1182, mediate the interaction with ERK2, and that the leucine zipper motifs of topoisomerase $II{\alpha}$ and $II{\beta}$ are not required for its physical binding to ERK2. Our results suggest that topoisomerase $II{\beta}$ residues 1099-1263, and topoisomerase $II{\alpha}$ residues 1078-1182, may be common binding sites for activator proteins.

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Chemical Modification of Extracellular Cytosine Deaminase from Chromobacterium violaceum YK 391

  • Kim, Tae-Hyun;Yu, Tae-Shick
    • Journal of Microbiology and Biotechnology
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    • 제8권6호
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    • pp.581-587
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    • 1998
  • Essential amino acids involved in the catalytic role of the extracellular cytosine deaminase from Chromobacterium violaceum YK 391 were determined by chemical modification studies. The enzyme activity required the reduced form of Fe (II) ion, since the enzyme was inhibited by ο-phenanthroline. The enzyme activity was completely inhibited by the chemical modifiers, such as p-chloromercuribenzoate (p-CMB), p-hydroxymercuribenzoate, and chloramine-T at 1 mM each. The enzyme activity was also markedly inhibited by pyridoxal-5'-phosphate, diethyl pyrocarbonate, and phenylmethylsulfonyl fluroride at 1 mM each. The inactivation of the enzyme activity with p-CMB was reversed by a high concentration of cytosine. Furthermore, the inactivation of the enzyme activity with p-CMB was also reactivated by 1 mM dithiothreitol, 1 mM 2-mercaptoethanol, 1 mM cysteine-HCI, 10% ethyl alcohol, and 10% methyl alcohol. These results suggested that cysteine and methionine residues might be located in or near the active site of the enzyme, while lysine, histidine, and serine residues might be indirectly involved in the enzyme activity.

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