• 제목/요약/키워드: 2-D gel electrophoresis

검색결과 336건 처리시간 0.034초

연어 뇌에서 N-Methyl-D-Aspartate 수용체 아단위 NR2A와 NR2B의 분포 (Distribution of N-Methyl-D-Aspartate Receptor Subunits NR2A and NR2B in Chum salmon Brain)

  • 진덕희;문일수
    • 생명과학회지
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    • 제9권6호
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    • pp.722-727
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    • 1999
  • 본 연구는 연어 뇌 연접의 단백질구성에 대한 기초연구로서, 기억형성에 중요한 역활을 하는 NMDA 수용체의 분포와 PTK에 의한 인산화에 대하여 조사하였다. 본 실험에 사용한 연어 뇌의 PSD 분획에서는 Coomassie로 염색할 경우 20여개 분명한 단백질띠를 확인할 수 있었으며, 소량으로 존재하여 전체적으로 도말되어 보이는 펩타이드의 수는 알 수 없었다. 이들 중 180kD 크기의 단백질은 phosphotyrosine 특이성 항체에 상대적으로 잘 인식이 되었다. 또한 이 180 kDa의 단백질들은 쥐에 있어서의 NR2A 및 NR2B의 위치인 분자량 약 180kD의 위치와 동일한 것으로 보아 연어에 있어서도 후각기능에 관계하는 NR2A 및 NR2B가 존재함을 추정할 수 있다.

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해양동물 구멍밤고둥의 렉틴 성분 연구 (Studies on Lectins from Marine Animal Chlorostoma argyrostoma turbinatum)

  • 정시련;최일식;전경희
    • 생약학회지
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    • 제25권2호
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    • pp.121-131
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    • 1994
  • Two kinds of new lectins, CATL-I and CATL-II, were partially purified from the intestine of Chlorostoma argyrostoma turbunatum by physical saline extraction, salt fractionation, ion exchange chromatography and gel filtration. CATL-I and CATL-II were purified 39.4 and 15.8 fold with a yield of 8.8 and 7.4%, respectively. On polyacrylamide gel electrophoresis, CATL-I demonstrated one major and one minor bands. This lectin agglutinated human and other animal erythrocytes nonspecifically and also agglutinated murine splenic lymphocytes. Carbohydrate specificity of the lectins was determined by inhibition of the agglutinability by methyl-${\alpha}-_D$-galactopyranoside and $_L-rhamnose$ at a final concentration of 6 mM. The lectins contained relatively high amounts of acidic amino acids, but the contents of sulfur containing amino acids were very low or was not estimated. Immunochemical studies were carried out to identify some properties of marine animal lectins.

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Proteomic Analysis of Proteins of Weissella confusa 31 Affected by Bile Salts

  • Lee, Kang Wook;Lee, Seung-Gyu;Han, Nam Soo;Kim, Jeong-Hwan
    • Journal of Microbiology and Biotechnology
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    • 제22권10호
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    • pp.1432-1440
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    • 2012
  • Weissella confusa 31, an isolate from human feces, possesses desirable properties as a probiotic strain, including bile salt resistance. W. confusa 31 is not inhibited by bile salts up to 0.3% concentration. Proteins affected by bile salts (0.05%) were examined by 2-D gel electrophoresis. Our proteomic analyses revealed that the intensities of 29 spots were changed, where 17 increased (including 2 spots observed only under the bile salts stress conditions) and 12 decreased. Proteins were identified by MALDI-TOF mass spectrometry. Proteins increased in the band intensities included adenylate kinase (12.75-fold increase), Clp-like ATP-dependent protease (11.91-fold), 6-phosphogluconate dehydrogenase (10.35-fold), and HSP 70 (5.07-fold). Some of the increased or decreased proteins are also known to be involved in other types of stress responses.

디젤분진이 체세포에서의 DNA 손상에 미치는 영향 (Genotoxic Effects of Diesel Exhaust Particle Extract in NIH/3T3 Cells)

  • 허찬;김남이;정규혁;문창규;허문영
    • Environmental Analysis Health and Toxicology
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    • 제19권4호
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    • pp.335-344
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    • 2004
  • Diesel exhaust particle (<2.5 ${\mu}{\textrm}{m}$, DEP$_{2.5}$) is known to be probarbly carcinogenic (IARC group 2A). DEP$_{2.5}$ contains organic compounds such as polycyclicaromatic hydrocarbon (PAH), heterocyclic compounds, phenols, and nitroarenes. Reactive oxygen species (ROS) are generated by DEP$_{2.5}$ without any biological activation system. Therefore, an alternative mechanism by which DEP$_{2.5}$ could be carcinogenic is known by the generation of oxidative DNA damage. The aim of this study was to investigate genotoxic effects of DEP$_{2.5}$ using single cell gel electrophoresis. In order to evaluate the mechanisms of DEP$_{2.5}$ genotoxicity, the rat micro-some mediated and DNA repair enzyme treated comet assays together with routine comet assay were performed. DEP$_{2.5}$ was collected from diesel engine bus and dichloromethane extract was obtained. The organic extract of DEP$_{2.5}$ revealed DNA damage itself in NIH/3T3 cells. And it showed both oxidative and microsome mediated DNA damages. Vitamin C as an model antioxidant reduced DNA damage in endonuclase III treated comet assay. One of flavonoid, galangin as a CYP1A1 inhibitor reduced DNA damage in the presence of S-9 mixture. Our results show that DEP$_{2.5}$ are genotoxic and a great source of oxidative stress, but antioxidants can significantly reduce oxidative DNA damages. And DEP$_{2.5}$ may contain indirect mutagens which can be inhibited by CYP inhibitors.d by CYP inhibitors.

Identification of novel $Ca^{2+}$ binding proteins in junctional sarcoplasmic reticulum of rabbit skeletal muscle

  • Jung, Dai-Hyun;Mo, Sang-Hyun;Kim, Do-Han
    • 한국생물물리학회:학술대회논문집
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    • 한국생물물리학회 2002년도 제9회 학술 발표회 프로그램과 논문초록
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    • pp.56-56
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    • 2002
  • Muscle contraction and relaxation are regulated by the sarcoplasmic reticulum (SR)-mediated $Ca^{2+}$ release and $Ca^{2+}$ uptake. The SR functions are closely related with the proteins residing in the SR such as ryanodine receptor, $Ca^{2+}$-ATpase, calsequestrin, triadin and junctin. In an effort to further identify important functional SR proteins, experiments of sucrose-density gradient of SR fractionation, concanavalin A treatment, 2D gel electrophoresis, $^{45}$ Ca$^{2+}$ overlay, Strains-all staining, and peptide finger printing (PFP) were carried out.(omitted)d)

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Stress Tolerance of Bifidobacterium infantis ATCC 27920 to Mild-heat Adaptation

  • Kang, Seok-U;Kim, Young-Hoon;Cho, In-Shick;Kang, Ja-Heon;Chun, Il-Byung;Kim, Kwang-Hyun;Oh, Se-Jong
    • Food Science and Biotechnology
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    • 제18권1호
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    • pp.249-252
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    • 2009
  • Two-dimensional gel electrophoresis (2-DE) was employed to assess the thermo-tolerance characteristics of Bifrdobacterium infantis ATCC 27920 to mild heat adaptation. When exposed to various heat levels, pH, and hydrogen peroxide ($H_2O_2$) stress conditions, B. infantis ATCC 27920 exhibited high level of stress resistance. Under mild-heat treatment ($46^{\circ}C$), no significant change in viability level was observed after 2 hr. Interestingly, improved viability was observed in mild-heat adapted ($46^{\circ}C$ for 1 hr) cultures exposed to $55^{\circ}C$, in comparison to control experiments. Viability was not affected by pH, bile, and $H_2O_2$ stress conditions. 2-DE analysis revealed those mild-heat adaptation up-regulated 4 proteins and down-regulated 3 proteins. Among these protein spots, isopropyhnalate dehydratase (leuD), glycosyltransferase (glgA), and ribosomal protein L5 (rp1E) were identified by matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALD1-TOF/MS).

Proteomic Analysis of the Hydrophobic Fraction of Mesenchymal Stem Cells Derived from Human Umbilical Cord Blood

  • Jeong, Ju Ah;Lee, Yoon;Lee, Woobok;Jung, Sangwon;Lee, Dong-Seong;Jeong, Namcheol;Lee, Hyun Soo;Bae, Yongsoo;Jeon, Choon-Ju;Kim, Hoeon
    • Molecules and Cells
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    • 제22권1호
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    • pp.36-43
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    • 2006
  • Mesenchymal stem cells (MSCs) are promising candidates for cell therapy and tissue engineering, but their application has been impeded by lack of knowledge of their core biological properties. In order to identify MSC-specific proteins, the hydrophobic protein fraction was individually prepared from two different umbilical cord blood (UCB)-derived MSC populations; these were then subjected to two-dimensional (2D) gel electrophoresis and peptide mass fingerprinting matrix-assisted laser desorption/ionization (MALDI)-time of flight (TOF)-mass spectrometry (MS). Although the 2D gel patterns differed somewhat between the two samples, computer-assisted image analysis identified shared protein spots. 35 spots were reliably identified corresponding to 32 different proteins, many of which were chaperones. Based on their primary sub-cellular locations the proteins could be grouped into 6 categories: extracellular, cell surface, endoplasmic reticular, mitochondrial, cytoplasmic and cytoskeletal proteins. This map of the water-insoluble proteome may provide valuable insights into the biology of the cell surface and other compartments of human MSCs.

한국산 초파리 집단의 유전 생화학적 연구: 노랑 초파리의 $\alpha$-Glycerophosphate dehydrogenase allele에 대하여

  • 정용재;한영수;정영란
    • 한국동물학회지
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    • 제25권3호
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    • pp.123-129
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    • 1982
  • 한국산 초파리집단의 유전적 특성을 생화학적으로 분석하여 분자유전학적 측면에서 집단 유전학적 체계를 확립하기 위하여 한국산 노랑초파리 (Drosophila melanogaster)의 11개 지역의 자연집단 (청량리, 등촌동, 신촌, 용산, 부천, 춘천, 대전, 대구, 광주, 나주 및 노화도)의 $\\alpha$-glycerophosphate dehydrogenase (\\alpha-GPDH) 인자형을 agarose gel 전기영동법에 의하여 조사 분석한 결과는 다음과 같다. 1. 한국산 노랑초파리 11가지 자연집단에서 $\\alpha$-GPDH allele는 상당한 다형현상을 이루고 있다. 2. $\\alpha$-GPDH allele의 heterozygosity는 $45\\sim50%$로서 상다히 높은 값을 보여주고 있다. 3. $\\alpha$-GPDH allele의 FF 인자형의 빈도는 SS인자형과 비슷하나 FS 인자형보다는 낮은 값을 보이고 있다. 4. $\\alpha$-GPDH allele의 F, S 두인자가 거의 비슷한 빈도를 보이고 있다.

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Biochemical Characterization of Serine Proteases with Fibrinolytic Activity from Tenodera sinensis (Praying Mantis)

  • Kim, Yeong-Shik;Hahn, Bum-Soo;Cho, So-Yean;Chang, Il-Moo
    • Toxicological Research
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    • 제17권
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    • pp.97-104
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    • 2001
  • Three types of proteases (MEF-1, MEF-2 and MEF-3) were purified from the egg cases of Ten-odera sinensis using ammonium sulfate fractionation, gel filtration on Bio-Gel P-60 and affinity chromatography on DEAE Affi-Gel blue gel. The proteases were assessed homogeneous by SDS-polyacrylamide gel electrophoresis and have molecular weight of 31,500, 32,900 and 35,600 Da, respectively. The N-terminal regions of the primary structure were compared and they were found to be different each other. MEFs readily digested the $A\alpha$ - and B$\beta$-chains of fibrinogen and more slowly the ${\gamma}$-chain. The action of the enzymes resulted in extensive hydrolysis of fibrinogen and fibrin, releasing a variety of fibrinopeptides. MEF-1 was inactivated by Cu$^{2+}$ and Zn$^{2+}$ and inhibited by PMSF and chymostatin. MEF-2 was inhibited by PMSF, TLCK. soybean trypsin inhibitor. MEF-3 was only inhibited by PMSF and chymostatin. Antiplasmin was not sensitive to MEF-1 but antithrombin III inhibited the enzymatic activity qf MEF-1. MEF-2 specifically bound to anti plasmin Among the chromogenic protease substrates, the most sensitive one to the hydrolysis of MEFs was benzoyl-Phe-Val-Arg-p-nitroanilide with maximal activity at pH 7.0 and 3$0^{\circ}C$. MEF-1 preferentially cleaved the oxidized B-chain of insulin between Leu15 and Tyr16. In contrast, MEF-2 specifically cleaved the peptide bond between Arg23 and Gly24. D-dimer concentrations increased on incubation of cross-linked fibrin with MEF-1, indicating the enzyme has a strong fibrinolytic activity.ity.

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콩나물 제조중(製造中) 질소화합물(窒素化合物)의 변화(變化)와 그 영양학적(營養學的) 연구(硏究) -제육보(第六報). 단백질(蛋白質)의 전기영동양상변화(電氣泳動樣相變化)- (Changes of Nitrogen Compounds and Nutritional Evaluation of Soybean Sprout -Part VI. Changes in electrophoretic pattern of protein-)

  • 양차범;박상기;윤석권;박훈
    • Applied Biological Chemistry
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    • 제27권2호
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    • pp.129-134
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    • 1984
  • 콩나물을 온도별로 길러 생장과정별 단백질조성을 7.5% polyacryl amide gel 전기영동법으로 조사하고 조제표품단백질의 그것으로 동정하였다. 원료대두(大豆)에서는 주(主) band 5개(a,b,c,d 및 p)와 미소 band 10개가 나타났고 4일자자엽(日字子葉)에서는 8개의 주(主) band와 (o band포함)와 11개 미소 band로 최고 band수(數)를 보였다. 배축(胚軸)에서는 자엽(子葉)에서 발견(發見)되는 band들이 모두 나타났으나 분리(分離)가 선명치 못하였다. 생육이 경과함에 따라 자엽(子葉)에서는 a band가 먼저, b,c,d band가 다음으로 감소해 갔으며 미소 band o+p는 6일(日)까지 증가하다 그후 감소하였다. 배축(胚軸)에서는 모든 band가 계속 증가하였는데 미소 band와 b+c+d가 증가속도가 컸다. a band는 7S, b+c+d는 11S, o+p는 2S globulin으로 동정되었다. 생육온도가 높아짐에 따라 자엽(子葉)에서의 감소와 배축(胚軸)에서의 증가속도가 빨라졌으며 7S보다 11S가 촉진되었다.

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