• 제목/요약/키워드: pH-gradient

검색결과 312건 처리시간 0.027초

Bence Jones 단백질(蛋白質)의 정제(精製) 및 N-말단검출(末端檢出) (Purification and N-Terminal Study of Bence Jones Proteins)

  • 김준평
    • Applied Biological Chemistry
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    • 제13권1호
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    • pp.59-64
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    • 1970
  • Bence Jones 단백질(蛋白質)(As,Ik,In)를 DEAE-Sephadex A-50 Column으로 0.02M phosphate Buffer(pH8.)와 0.5M NaCl를 gradient로 사용(使用)해 정재(精製)하였다. 시료(試料) As(K-형(型))의 정재결과(精製結果) 주성분(主成分)인 F-1는 조단백질(粗蛋白質) 500mg로부터 350mg 얻었으며 다른 시료(試料) Im와 Ik($lambda$-형(型))에서도 각각 주성분(主成分)을 242mg(Im)와 146mg(Ik) 얻을 수 있었다. 정재(精製)한 시료(試料)는 그의 순도(純度)를 알기 위해 Poly acryl amide의 Disc electrophoresis로 확인한 후 K-형(型) Bence jones단백질(蛋白質)의 일종(一種)인 As와 $lambda$-형(型)의 일종(一種)인 IM를 6N-HCI로 $105^{\circ}C$에서 20시간(時間) 가수분해(加水分努)후 Amino 산(酸) 조정(組成)을 살펴보았다. DNP법(法)으로 시료(試料) K-형(型)(As, Ko, Ta)의 단백질(蛋白質)의 N-말단(末端)을 검출(檢出)해 Ta는 glutamic acid, Ko, As는 Aspartic acid임을 확인하였으며 Paper 상(上)에 나타난 DNP-Amino 산(酸)은 5% $NaHCO_3$ (pH8) 4ml에 추출(抽出)해 그 수량(收量)을 ${\varepsilon}=18.1{\times}10^3DNP$ $Asp\;{\varepsilon}=17.41{\times}10^(3){\;}DNP{\;}Glu$에 의(依)해 산출(算出)하였더니 Ko는 54.3%, As는 65%이였으며 Ta는 85%의 수량(收量)이였다. ${\lambda}$-형(型)의 Im,Ik의 시료(試料)도 DNP법(法)으로 N-말단(末端)을 검출(檢出)해 보았더니 검출(檢出)되지 아니하였다.

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Cobra (Ophiphagus hannah) 독으로부터 세포 독성 단백질의 분리

  • 김영식;이병무;안미영;박재갑;현진원;임경화;박호군
    • 한국응용약물학회:학술대회논문집
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    • 한국응용약물학회 1994년도 춘계학술대회 and 제3회 신약개발 연구발표회
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    • pp.205-205
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    • 1994
  • 동물, 곤충, 해조류 등으로 부터 얻어진 독소는 해로운 것으로 알려져 있지만 의학적으로 유용한 화합물을 내포하고 있다. 본 연구는 이러한 점에 착안하여 독사, 또는 곤충의 독 30여종을 대상으로 위암 세포인 SNU-1에 대해서 MTT assay에 의한 세포 독성 시험을 실시하였다. 이 중 세포 독성 활성이 가장 놓은 코브라 계통인 Ophiphagus hannah의 독으로 부터 세포 독성 물질의 정제를 시도하였다. HPLC-GPC와 HPLC-lEX 또는 RP-HPLC를 이용하여 정제하였고 각 분획들을 암세포주에 대해서 시험하였다. TSK-2000SW로 부터 2개의 분획을 얻어 분획 I에서 $IC_{50}$/ 의 값이 0.5 - 3 $\mu\textrm{g}$/ml의 범위 내에서 확인되었다. 분자량이 작은 분획 II에서는 세포독성이 작게 나타났지만 농축하여 성분 분석을 시도하였다. 분획 I을 RP-HPLC를 이용하여 TFA와 acetonitrile의 linear gradient에 의해 더욱 분리를 하였고, phopholipase $A_2$ 활성의 가능성도 측정하였다. 분획 II는 Mono S 칼람을 이용하여 ammonium acetate농도에 따른 pH gradient에 의한 분리를 시도하여 순도를 SDS-PAGE에 의해서 확인하였다. 각각의 크로마토그리피로 부터 얻은 분획에 대해서 세포 독성을 실시중에 있으며 성질도 동정중에 있다.

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The dynamic instability of FG orthotropic conical shells within the SDT

  • Sofiyev, Abdullah H.;Zerin, Zihni;Allahverdiev, Bilender P.;Hui, David;Turan, Ferruh;Erdem, Hakan
    • Steel and Composite Structures
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    • 제25권5호
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    • pp.581-591
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    • 2017
  • The dynamic instability of truncated conical shells subjected to dynamic axial load within first order shear deformation theory (FSDT) is examined. The conical shell is made from functionally graded (FG) orthotropic material. In the formulation of problem a dynamic version of Donnell's shell theory is used. The equations are converted to a Mathieu-Hill type differential equation employing Galerkin's method. The boundaries of main instability zones are found applying the method proposed by Bolotin. To verify these results, the results of other studies in the literature were compared. The influences of material gradient, orthotropy, as well as changing the geometric dimensions on the borders of the main areas of the instability are investigated.

계 골격근에서 15S ATPase의 순수분리 및 특성연구 (Purification and Characterization of a 15S Arpase from Chick Skeletal Muscle)

  • 심규석;채광수
    • 한국동물학회지
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    • 제37권4호
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    • pp.524-530
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    • 1994
  • An Arpase complex has been purified to apparent homogeniety from the extract of chick skeletal muscle using conventional column chromatographies and glycerol density gradient centrifugation. This eWe has a sedimentation coefficient of 15S as determined by the gradient centrifugation and therefore is referred to as the 15S ATPase. It behaves as a 600-kOa molecule upon gel filtration analysis using a Superose-6 column. However, the ATPase runs as a 95-kDa polvpeptide when analyzed by polvacrvlamide gel electrophoresis in the presence of sodium dodecyl sulfate. Thus, the Arpase is likely to consist of six identical subunits of 95 KDa. It has a Km value of 0.6 mM for ATP and is maximally active at pH 9.

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Jn vitro에서 cortisol이 개 말초혈액 단핵구세포의 증식에 미치는 영향 (Ju vitro Effect of Cortisol on the Proliferation of Canine Peripheral Blood Mononuclear Cells)

  • 나기정;양만표
    • 한국임상수의학회지
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    • 제14권2호
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    • pp.230-234
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    • 1997
  • In vitro effect of cortisol on the proliferation of canine peripheral blood mononuclear cells (MNC) was examined. The MNC was isolated from peripheral blood by a gradient centrifugation with Picoll-Hypaque. The cell proliferation assayed using a noneradioactive 5-Bromo-2'-deoxy-uridine (BrdU) kit. The MNC proliferated well in response to either phrtobeRagg$]$utinin-p (PHA-P) or culture supernatant from MNC stimulated with PHA-p. However, these proliferative responses of MNC were not affected by addition of coitisol of 1 to 1,OOfl ng/ml. The addition of cortisol in MNC culture with either PHA-P or corture supernatBnt from MNC stimulated with PHA-P far 4 days wag not also influenced on the viabilities of cultured MNC. In conclusions it was able to assay the cell proliferation with BrdU instead of radioactive isotope e.g. tritiated thymidine (3H-TdR). These results suggested that cortisol does not at least influence on MNC proliferation in vitro.

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Abundance of Polychaetes in Lake Shihwa, Korea

  • Lee, Jong-Hyeon;Koh, Chul-Hwan
    • Journal of the korean society of oceanography
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    • 제34권2호
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    • pp.122-131
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    • 1999
  • This study examined the relationship between the sediment pollution and the occurrence of polychaetes in a heavily polluted saltwater lake, Lake Shihwa on the west coast of Korea, separated from the sea by a dike in 1994. The species composition of polychaete assemblage was compared with that found off the lake in Kyeonggi Bay. Environmental variables investigated both in and off the lake were the grain size distribution, chemical oxygen demand(COD) and metal concentrations (Al, Fe, Mn, V, Co, As, Pb, Cr and Cu) in the sediment. We sampled sediments at 10 stations in the lake and 25 stations in Kyeonggi Bay using a modified van Veen grab. The levels of COD, chromium, and copper in sediments were much higher in Lake Shihwa than in Kyeonggi Bay. Differences in the species composition were found along the pollution gradient. An azoic zone was observed in the most heavily polluted area at the upper reach of the lake and the Polydora ligni zone in the center of the lake. Bottom fauna were diverse in Kyeonggi Bay; however, Heteromastus filiformis predominated in the organically enriched sediment. The density of dominant species differed along the pollution gradient. The highest density or H. filiformis was round at the COD level or around 5.8 mg/g. The COD level in the sediment where P. ligni predominated was tow-fold higher than that where H. filiformis occurred in large numbers. The chromium and copper contents at which P. ligni showed a maximum abundance were 120 mg/kg and 127 mg/kg, respectively. The density of H. filiformis was highest at concentrations of 56 mg/kg chromium and 13 mg/kg copper.

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HPLC Separation of Isoquinoline Alkaloids for Quality Control of Corydalis species

  • Kim, Eun-Kyung;Jeong, Eun-Kyung;Han, Sang-Beom;Jung, Jee-H.;Hong, Jong-Ki
    • Bulletin of the Korean Chemical Society
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    • 제32권10호
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    • pp.3597-3602
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    • 2011
  • A simple and rapid analytical method was developed for the determination of eight isoquinoline alkaloids in Corydalis species. Eight isoquinoline alkaloids, including 2 aporphine alkaloids (isocorydine and glaucine) and 6 protoberberine alkaloids (coptisine, palmatine, berberine, canadine, corydaline, and tetrahydrocoptisine) were used as chemical markers which have a various biological activity and determined for quality control of Corydalis (C.) species (C. ternata, C. yanhusuo, and C. decumbens). To evaluate the quality of these herbal medicines, LC chromatographic separation of alkaloids were preferentially investigated on reversed-phase C18 column with pH variation and composition of mobile phase. In addition, the separation of these alkaloids in herbal extracts was found to be significantly affected on mobile phase composition using gradient elution. Especially for C. yanhusuo extract, berberine was seriously interfered with other alkaloid extracted from sample matrix when mobile phase composition was not optimized. As results, these compounds were successfully separated within 28 min using 10 mM ammonium acetate containing 0.2% triethylamine (adjusted at pH 5.0) as a mobile phase with gradient elution. On the basis of optimized HPLC conditions, 23 different Corydalis species samples were analyzed for the determination of alkaloid levels. In addition, principal component analysis (PCA) combined with the chromatographic data could be successfully classified the different geographic origin samples.

Polymorphisms of SLC22A9 (hOAT7) in Korean Females with Osteoporosis

  • Ahn, Seong Kyu;Suh, Chang Kook;Cha, Seok Ho
    • The Korean Journal of Physiology and Pharmacology
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    • 제19권4호
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    • pp.319-325
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    • 2015
  • Among solute carrier proteins, the organic anion transporters (OATs) play an important role for the elimination or reabsorption of endogenous and exogenous negatively charged anionic compounds. Among OATs, SLC22A9 (hOAT7) transports estrone sulfate with high affinity. The net decrease of estrogen, especially in post-menopausal women induces rapid bone loss. The present study was performed to search the SNP within exon regions of SLC22A9 in Korean females with osteoporosis. Fifty healthy controls and 50 osteoporosis patients were screened for the genetic polymorphism in the coding region of SLC22A9 using GC-clamped PCR and denaturing gradient gel electrophoresis (DGGE). Six SNPs were found on the SLC22A9 gene from Korean women with/without osteoporosis. The SNPs were located as follows: two SNPs in the osteoporosis group (A645G and T1277C), three SNPs in the control group (G1449T, C1467T and C1487T) and one SNP in both the osteoporosis and control groups (G767A). The G767A, T1277C and C1487T SNPs result in an amino acid substitution, from synonymous vs nonsynonymous substitution arginine to glutamine (R256Q), phenylalanine to serine (F426S) and proline to leucine (P496L), respectively. The Km values and Vmax of the wild type, R256Q, P496L and F426S were 8.84, 8.87, 9.83 and $12.74{\mu}M$, and 1.97, 1.96, 2.06 and 1.55 pmol/oocyte/h, respectively. The present study demonstrates that the SLC22A9 variant F426S is causing inter-individual variation that is leading to the differences in transport of the steroid sulfate conjugate (estrone sulfate) and, therefore this could be used as a marker for certain disease including osteoporosis.

환경구배에 따른 바닷새 번식지 토양환경 특성 (Soil Environmental Characteristics According to the Environmental Gradient in Seabird Breeding Sites)

  • 김진석;임병선;박정원;명현호
    • 생태와환경
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    • 제46권1호
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    • pp.18-24
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    • 2013
  • 본 연구는 바다새 서식지 복원 및 식생복원을 위해 2011년 4월부터 12월까지 전라남도 신안군 칠발도와 소구굴도지역에 대한 식물군락 및 환경구배에 따른 바닷새 번식지 토양환경 요인을 분석하였다. 토양요인을 분석한 결과 칠발도는 닭의장풀군락이 수분함량, 유기물함량, 염분농도, 총질소가 높게 나타났으며, 쇠무릎 군락은 낮게 나타났다. 또한, 바닷새 서식처로 이용되는 밀사초 군락은 토양환경요인 분포범위가 넓고 유입종인 쇠무릎군락과 쑥군락은 좁게 나타났다. 소구굴도 밀사초군락 토양pH는 3.14로 가장 낮았고, 갯기름나물군락은 3.84 가장 높게 나타났다. 수분함량과 유기물함량은 갯기름나물군락에서 가장 높게 나타났으며, 쇠무릎군락이 35.9%로 가장 낮게 분석되었다. 염분농도와 총질소는 밀사초군락이 가장 높게 나타났다. 고도에 따른 토양환경특성에서는 칠발도의 경우 고도가 높아질수록 수분함량, 유기물함량, 염분농도 및 총질소가 낮아지는 유의성을 보였다. 소구굴도 토양수분함량은 평균 35.73%, 유기물함량 16.38%, pH 3.34로 전체적으로 산성토양이었다. 토양산성화는 조류분비물에 의해 영향을 받았을 것으로 판단된다. 염분농도는 평균 2.11 ppt, 총질소는 평균 $6.43mg\;g^{-1}$로 분석되었다. 소구굴도는 섬면적이 좁고, 고도가 낮아 전체적으로 토양환경요인의 분석결과는 크게 차이가 나타나지 않았으나, 수분함량은 환경구배에 대해 고도에 의한 유의성을 보였다.

Proteome Approach as a Tool for the Efficient Separation of Seed Storage Proteins from Buckwheat

  • Cho, Seong-Woo;Kwon, Soo-Jeong;Roy, Swapan Kumar;Woo, Sun-Hee
    • 한국작물학회지
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    • 제60권1호
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    • pp.29-32
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    • 2015
  • Two-dimensional electrophoresis (2-DE) was executed to separate the seed storage proteins from the buckwheat. The proteins extracted from the whole seed proteins were better separated and observed in the use of lysis buffer. Using this method, the highly reproducible isoelectric focusing (IEF) can be obtained from polyacrylamide gels, and IEF from the polyacrylamide gel at all the possible pH range (5.0-8.0) was more easily separated than IPG (immobilized pH gradient) gels. The polyacrylamide gels in the first dimension in 2-DE was used to separate and identify a number of whole seed proteins in the proteome analysis. In this new apparatus using 2-DE, 27cm in length of plate coated with polyacrylamide gel was used and the experiment was further investigated under the various conditions.