• Title/Summary/Keyword: polypeptides

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Identification and Partial Purification of Two Hydrogenase Isoenzymes from Escherichia coli (대장균으로부터 두 종류의 수소발생 동위효소의 확인과 부분정제)

  • 최석정;양철학
    • Korean Journal of Microbiology
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    • v.29 no.5
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    • pp.296-300
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    • 1991
  • The membrane-bound Escherichia coli hydrogenases were purified partially by the solubilization with detergents. the E. coli crude extract was solubilized with sodium deoxycholate and dialyzed against the buffer containing Triton X-100. Two different hydrogenases were obtained by the DEAE-cellulose, hydroxyapatite and Sephedex G-200 column chromatography. The one was unstable during purification and contained 70- and 47-kDa polypeptides as major proteins. The other showed high H2-evolving activity and had major polypeptides of Mr 31 and 27. Those polypeptides were detected by the two-dimensional electrophoresis.

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Studies on Inactivation and Reactivation of Isolated Photosystem II Complexes in Spinach (시금치에서 분리한 광계 2 복합체의 불활성화와 재활성화에 대한 연구)

  • 전현식
    • Journal of Plant Biology
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    • v.33 no.4
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    • pp.277-283
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    • 1990
  • Inactivation and reactivation of photosynthetic oxygen evolving complex were studied with isolated spinach (Spinacia oleraceda. L.) photosystem II particles by the activity of oxygen evolution and chlorophyll fluorescence. When the particles were treated with Tris and urea, the oxygen evolution was inactivated and three polypeptides having molecular weights of 33 kDa, 24 kDa and 18 kDa were simultaneously released. But in NaCl-treated particles, two polypeptides of 24 kDa and 18 kDa were removed from PS II particles. The oxygen evolution activities of Tris and urea-treated particles were not restored by adding cation ions (Mg2+, Mn2+ and Ca2+), but the NaCl-treated particles were restored by exogenously added Ca2+. The removal of these extrinsic polypeptides, especially 33 kDa, markedly showed the decrease of the variable fluorescence (Fv). These results are likely to be due to dissipate thermal energy by antenna of photosystem II complexes.

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SDS-polyacrylamide Gel Electrophoresis on Partially Purified Ginseng Proteins (인삼단백질분획에 대한 SDS-폴리 아크릴아미드 전기영동)

  • 김춘미
    • YAKHAK HOEJI
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    • v.27 no.4
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    • pp.315-320
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    • 1983
  • Ginseng proteins have been extracted and partially purified by the methods of ammonium sulfate fractionation, heat inactivation and Sephadex G-75 column chromatography. The final three fractions obtained (GI, GII and GIII) were subjected to paper chromatography and SDS-polyacrylamide gel electrophoresis. Molecular weights of polypeptide chains from each fraction were est-mated by the standard line obtained from the plot of electrophoretic mobilities against the logarithm of molecular weights of standard proteins. Results are as follows: 1) GI showed three protein spots and four polypeptides of which M.W. were 63,000, 60,000, 56,000, 51,000 and 34,200. 2) GII showed four protein spots and five polypeptides with their M.W. of 64,600, 56,000, 45,400, 35,800, end 25,200, 3) GIII showed three protein spots and four polypeptides with their M.W. of 66,000, 63,000, 56,000 and 22,000.

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Comparative study on the properties of polypeptides induced by NaCl, drought and temperature treatments in rice seedlings (NaCl, 한발 및 온도 처리에 따른 유묘기 수도의 폴리펩티드 속성의 비교분석)

  • Lim, Gum-Chun;Jung, Yeoung-Sang;Shin, Jeong-Sheop
    • Applied Biological Chemistry
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    • v.35 no.6
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    • pp.485-489
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    • 1992
  • Plants are altered not only in the outward appearance but also in their physiological and biochemical properties with reaction to the environmental stresses; particularly, the biosynthetic system of protein in situ rapidly responds to this. In order to investigate the change of properties of polypeptides in rice plants induced by several stresses, the seedlings were subjected to exposure to NaCl, drought, and low and high temperatures, respectively, and then some aspects of polypeptide variations were compared. Without exception, the rice plant, which is somewhat tolerant to environmental change, shows the alteration in several polypeptides. Moreover, newly synthesized polypeptides were observed in response to stresses. The existing proteins for the primary metabolic pathways were markedly decreased as each treatment progressed.

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Fused Polypeptide with DEF Induces Apoptosis of Lung Adenocarcinoma Cells

  • Liang, Ai-Ling;Zhang, Ting-Ting;Zhou, Ning;Huang, Di-Nan;Liu, Xin-Guang;Liu, Yong-Jun;Tu, Zhi-Guang
    • Asian Pacific Journal of Cancer Prevention
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    • v.14 no.12
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    • pp.7339-7344
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    • 2013
  • To analyze the effects of a new unknown peptide DEF on the growth of tumor cells, a fused polypeptide TAT-DV1-DEF was designed and synthesized. The lung adenocarcinoma cell line GLC-82 treated with TAT-DV1-DEF was analyzed with a cell counting kit 8, and the location of polypeptides in cells was observed under laser confocal microscopy. The efficiency of polypeptide transfection and changes in nuclear morphology were analyzed by flow cytometry and fluorescence microscopy, respectively. Finally, the mechanism of tumor cell growth inhibition was evaluated by Western blotting. We found that TAT-DV1-DEF could significantly inhibit the growth of the lung adenocarcinoma cell line GLC-82, but not the normal human embryonic kidney cell line HEK-293. Polypeptides were found to be mostly localized in the cytoplasm and some mitochondria. The efficiency of polypeptide transfection in the two cell types was approximately 99%. Apoptotic nuclei were observed under fluorescence microscopy upon treatment with polypeptides and DAPI staining. Western blot analyses indicated that the polypeptide inhibition of tumor cell growth was apoptosis dependent. In the present study, we demonstrated that fused polypeptides could induce apoptosis of the lung adenocarcinoma cell line GLC-82, indicating that the new unknown peptide DEF has antitumor effects.

LHCP phosphorylation and Chlorophyll-Fluorescence Quenching of PSII in Ginseng Thylakoid Members (인삼 틸라코이드에서 광계II의 LHCP 인산화와 형광 Quenching)

  • 양덕조;김명원
    • Journal of Ginseng Research
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    • v.16 no.2
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    • pp.124-128
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    • 1992
  • Vsing the phosphorylated thylakoid membrane induced by 5-35kLux of light intensities, we investigated the chlorophyll nuorescence quenching of PSII and the phosphorylation of LHCPII i in relation to the chlorophyll-bleaching of Panax ginseng C.A. Meyer. In the Presence of DCMU, both of the fluorescence yield of non-phosphorylated thylakoid and the rate of fluorescence quencing dependent on the Phosphorylation were high p. ginseng more then Glyine max L. And at the high light F intensity (above 25 kLux) the fluorescence quenching rate of p. ginseng compared with that of G. max reached nearly to 2 times. The LHCPII of P. ginseng was composed of 3 major Polypeptides (24.5, 26 and 27kD) and 3 minor polypeptides (24, 25.3 and 28.3kD) in the region of 24-29kD and differed, from G. max in both of the number and quantity of polypeptides. Among these polypeptides, the phosphorylated polypeptide dependent on the light intensity was 24kD in P. ginseng.

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Roles of growth factors, calcitonic polypeptides and neuropeptides in bone metabolism, osteoporosis and rheumatis arthritis

  • Lee, Tae-Kyun;Kim, June-ki;Kim, Kap-Sung;Chang, Jun-Hyuk;Jeong, Ji-cheon;Nam, Kyung-Soo;Kim, Cheorl-Ho
    • The Journal of Dong Guk Oriental Medicine
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    • v.9
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    • pp.1-23
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    • 2000
  • Osteoporosis is a common disorder characterized by reduced bone mineral density, deterioration of the microarchitecture of bone tissue and increased risk of fracture. The aim of treatment of osteoporosis is to maintain and, ideally, to restore bone strength safely. In recent years the role of polypeptide growth factors in bone metabolism has begun to appear. It has been proposed that alterations in the expression or production of growth factor can modulate the proliferation and activity of bone forming cells. Thus, the role of structurally diverse peptides for the management and diagnosis of osteoporosis has attracted the attention of many investigators. This paper reviews numerous findings concerning the use of polypeptides, hormones, and growth factors, for the management of osteoporosis. Many of the compounds mentioned here are experimental prototypes of new therapeutic classes. Though it is unlikely that some of the compounds may ever be used clinically, development of safe and efficacious agents in each class will define the future course of therapy for osteoporosis.

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Purification and Characterization of Vitellin from the Red Flour Beetle, Tribolium castaneum Herbst

  • Kim, Seong-Ryul;Choo, Young-Moo;Lee, Seong-Jin;Jin, Byung-Rae;Kim, Jeong-Ho;Heo, In-Bum;Shon, Hung-Dae
    • International Journal of Industrial Entomology and Biomaterials
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    • v.2 no.1
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    • pp.55-59
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    • 2001
  • The vitellin of the red flour beetled Tribolium castaneum Herbst was purified and characterized. The vitellin of T. castaneum was purified by the FPLC techniques, anion exchange chromatography and gel permeation chromatography. In native-polyacrylamide gel electrophoresis, vitellin of T. castaneum was detected as a single band. This native vitellin has molecular weight of 440 kDa. The vitellin of T. castaneum is composed of three polypeptides, designated Vnl (178 kDa), Vn2 (168 kDa) and Vn3 (52 kDa) in SDS-polyacrylamide gel electrophoresis. Three subunits of vitellin were presented in the female adult hemolymph and egg extracts, but not observed in the male. These three polypeptides gradually decreased during embryogenesis. Polyclonal antiserum raised against purified vitellin reacted with the three polypeptides, Vnl, Vn2 and Vn3. Antisera raised against Vn1 and Vn2 cross-reacted with the two large subunits, Vnl and Vn2, respectively. Another subunits Vn3, however, was not cross-reacted with these two antisera. Also, antiserum raised against Vn3 did not cross-react with the Vn1 and Vn2.

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Production of Repetitive Polypeptides for an Efficient DNA Analysis on a Microchip (Microchip상에서 효율적인 DNA 분석을 위한 반복단위 단백질의 생산)

  • Yi, Hyeon-Jin;Choi, Seok-Jin;Seo, Tae-Seok;Won, Jong-In
    • KSBB Journal
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    • v.25 no.2
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    • pp.199-204
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    • 2010
  • We generated the feasibility of DNA separation in free-solution using genetically engineered repetitive polypeptides as drag-tags. Two different-sized repetitive polypeptides were designed, expressed in E. coli, and purified. They were conjugated to a fluorescently labeled DNA (100 base), and the electrophoretic mobilities of these conjugate molecules were analyzed on a microchip. The results of these studies indicate that genetically engineered repetitive polypeptide is a prominent candidate for rapid and high-throughput genetic mutation detection, such as SNP analysis.

STRUCTURAL ANALYSIS OF THE SOLUBLE FORM OF BOVINE DOPAMINE BETA-HYDROXYLASE

  • Hwang, On-You;Joh, Tong-Hyub
    • Toxicological Research
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    • v.6 no.1
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    • pp.99-107
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    • 1990
  • Dopamine beta-hydroxylase (DBH) is a neurotransmitter biosynthetic enzyme which catalyzes the conversion of dopamine to norepinephrine. Although structural studies of the mature form of this enzyme have been extensive, the culmination of these finding had been hightly controversial and contradictory. In this study, biochemical approaches were taken to characterize the structure of mature DBH. Soluble bovine DBH was purified from aderenal medulla. Three bands of 69 kDa, 72 kDa and 75 kDa which were physically separable and similar in structure were observed by SDS-PAGE. Furthermore, gas phase sequence analysis revealed that the 72 kDa band consists of two polypeptides which are present at equimolar concentrations and differed in that one had three extra amino acids at the N-terminus. Taken together, the soluble form of DBH exist in at least four forms, three identified by SDS-PAGE, one of which consists of two polypeptides as identified by N-terminal sequence analysis. The significance of these forms and their possible biosynthetic mechanisms are discussed.

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