• 제목/요약/키워드: polypeptides

검색결과 232건 처리시간 0.031초

Formation of Cross-Linked Products of The Reaction Center D1 Protein in Photosystem II under Light Stress

  • Uchida, Suguru;Kato, Yoji;Yamamoto, Yasusi
    • Journal of Photoscience
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    • 제9권2호
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    • pp.382-384
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    • 2002
  • When illuminated with strong visible light, the reaction center Dl protein of photo system II is photodamage and degraded. Reactive oxygen species and endogenous cationic radicals generated by photochemical reactions are the cause of the damage to the Dl protein. Recently we found that the photodamaged Dl protein cross-links with the surrounding polypeptides such as D2 and CP43 in photosystem II. As the cross-linking reaction is dependent on the presence of oxygen, reactive oxygen species are suggested to be involved. Among the reactive oxygen species examined, ? OH was most effective in the formation of the cross-linked products. These results indicate that the cross-linking is mostly due to ? OH generated at photosystem II. The cross-linking site of the Dl protein is not known. As several tyrosine residues exist at the D­E loop of the Dl protein, there is a possibility that di-Tyr is formed between the D­E loop of the Dl protein and surrounding polypeptides during the strong illumination. Therefore, we examined the formation of di-Tyr using the monoclonal antibody against di-Tyr under excess illumination of the photosystem II membranes. The results obtained here suggest that no di-Tyr is formed during the excess illumination of photosystem II.

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Isolation and characterization of fowl adenovirus serotype 4 from chickens with hydropericardium syndrome in Korea

  • Park, Hong-Su;Lim, Il-Soo;Kim, Sang-Kyu;Kim, Toh-Kyung;Yeo, Sang-Geon
    • 대한수의학회지
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    • 제51권3호
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    • pp.209-216
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    • 2011
  • Four strains of fowl adenovirus (FAdV) were isolated from 4 flocks of broiler or layer chickens affected by hydropericardium syndrome in Korea. These FAdVs were classified as serotype 4 by restriction fragment length polymorphism patterns of hexon genes and whole genomes. The virus exhibited cytopathic effects consisting of rounding, ballooning and clustering in primary chicken embryo liver cell cultures. In transmission electron microscopy, virus particles in hexagonal shape aggregated exclusively in the nuclei of hepatocytes of the chickens as the typical appearances in adenovirus infections. Buoyant density of the virus in cesium chloride (CsCl) was 1.34 g/mL. The virus was stable to chloroform, ether, 50~70% ethanol, acidic condition at pH 3, 0.25% trypsin (1 : 250), heat at $50^{\circ}C$ for 30 min, but labile to 100% ethanol, heat at $52{\sim}60^{\circ}C$ for 30 min, 1 M $MgCl_2$ at $50^{\circ}C$ for 1 h, 1 : 2,000 formalin (37%). All of the physicochemical properties pertained to the characteristics of adenoviruses. Eight viral polypeptides were determined in CsCl-purified virus by sodium dodecyl sulfate-polyacrylamide gel electrophoresis.

Regulation of Chlorophyll-Protein Complex Formation and Assembly in Wheat Thylakoid Membrane

  • Guseinova, I.M.;Suleimanov, S.Y.;Aliev, J.A.
    • BMB Reports
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    • 제34권6호
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    • pp.496-501
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    • 2001
  • Lincomycin, an inhibitor of plastid protein synthesis, was found to block the synthesis of apoprotein P700 with a molecular mass of 72 kDa and the assembly of the Chl a-protein of PS I. Synthesis of the polypeptides of 48, 43.5, and 32 kDa of the PS II complex is also suppressed. This process is accompanied by the disappearance of the PS Two reaction center Chl a at 683 nm, and of the PS One reaction center Chl a at 690, 696, and 705 nm on the fourth derivative of the absorption spectra at 77K. Lincomycin does not affect the synthesis of LHC subunits. It increases the content of the two main Chl forms of LHC at 648 nm (Chl b) and 676 nm (Chl a). The low-temperature fluorescence ratio F736/F685 is also increased. However, the effect of cycloheximide (an inhibitor of cytoplasmic protein synthesis) leads to the reduction of polypeptides of the light-harvesting Chl a/b-protein complex in the range of 29.5-22 kDa. Under these conditions, the relative amount of Chl b and the F736/ F685 fluorescence ratio decrease significantly. This is obviously the result of blocking the LHC I and LHC II synthesis. At the same time rifampicin and actinomycin D (inhibitors which block transcription in chloroplast and nuclear genome, respectively) inessentially affect the characteristics of these complexes.

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Characterization of Fibrinolytic Proteases from Gloydius blomhoffii siniticus Venom

  • Choi, Suk-Ho
    • 대한약침학회지
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    • 제14권3호
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    • pp.71-79
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    • 2011
  • Objectives : This study was undertaken to identify fibrinolytic proteases from Gloydius blomhoffii siniticus venom and to characterize a major fibrinolytic protease purified from the venom. Methods : The venom was subjected to chromatography using columns of Q-Sepharose and Sephadex G-75. The molecular weights of fibrinolytic proteases showing fibrinolytic zone in fibrin plate assay were determined in SDS-PAGE (Sodium dodecyl sulfate-polyacrylamide gel electrophoresis) The effects of inhibitors and metal ions on fibrinolytic protease and the proteolysis patterns of fibrinogen, gelatin, and bovine serum albumin were investigated. Results : 1) The fibrinolytic fractions of the three peaks isolated from Gloydius blomhoffii siniticus venom contained two polypeptides of 46 and 59 kDa and three polypeptides of 32, 18, and 15 kDa and a major polypeptide of 54 kDa, respectively. 2) The fibrinolytic activity of the purified protease of 54 kDA was inhibited by metal chelators, such as EDTA, EGTA, and 1,10-phenanthroline, and disulfhydryl-reducing compounds, such as dithiothreitol and cysteine. 3) Calcium chloride promoted the fibrinolytic activity of the protease, but mercuric chloride and cobalt(II) chloride inhibited it. 4) The fibrinolytic protease cleaved preferentially A${\alpha}$-chain and slowly B${\beta}$-chain of fibrinogen. It also hydrolyzed gelatin but not bovine serum albumin. Conclusions : The Gloydius blomhoffii siniticus venom contained more than three fibrinolytic proteases. The major fibrinolytic protease was a metalloprotease which hydrolyzed both fibrinogen and gelatin, but not bovine serum albumin.

야생 흰 제비꽃(Viola patrinii DC.) 배양세포의 생성과정 중 단백질 및 아미노산의 함량변화 (Alterations in Protein and Amino acid Contents During Growth on Culture Cells of Wild Viola (Viola patrinii DC.))

  • 정용모;임현희;조영수;정정한;이재헌;서정해;권오창
    • 식물조직배양학회지
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    • 제27권3호
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    • pp.181-184
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    • 2000
  • Friable callus와 compact callus를 배양 단계별로 단백질을 정량한 결과 friable callus가 가장 낮았고 배양 0주째부터 단백질 함량이 서서히 증가하여 배양 3주째 최대함량이 되었으며, 4주째부터는 감소하기 시작했다. 전기영동법으로 단백질의 패턴을 조사한 결과 compact callus와 friable callus간에 특징적인 차이를 보이는 단백질 밴드가 관찰되었다. Friable callus는 약 28 KD, 31 KD과 35 KD에서 특이적으로 나타났으며. compact callus에서는 30 KD부근의 밴드에서 많은 변화가 일어났다. 전체 단백질의 구성 아미노산은 배앙 2∼3주에 가장 높은 함량을 보였고, friable callus에서 그 함량이 가장 낮았다.

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A quantitative method for detecting meat contamination based on specific polypeptides

  • Feng, Chaoyan;Xu, Daokun;Liu, Zhen;Hu, Wenyan;Yang, Jun;Li, Chunbao
    • Animal Bioscience
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    • 제34권9호
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    • pp.1532-1543
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    • 2021
  • Objective: This study was aimed to establish a quantitative detection method for meat contamination based on specific polypeptides. Methods: Thermally stable peptides with good responses were screened by high resolution liquid chromatography tandem mass spectrometry. Standard curves of specific polypeptide were established by triple quadrupole mass spectrometry. Finally, the adulteration of commercial samples was detected according to the standard curve. Results: Fifteen thermally stable peptides with good responses were screened. The selected specific peptides can be detected stably in raw meat and deep processed meat with the detection limit up to 1% and have a good linear relationship with the corresponding muscle composition. Conclusion: This method can be effectively used for quantitative analysis of commercial samples.

Alachlor에 의한 귀리 근단(根端) 분열조직(分裂組織)의 단백질(蛋白質) Pattern의 변이(變異) (Changes in the Polypeptide Patterns of Oat Root Tips Exposed to Alachlor)

  • 권성환;박기인;김재철
    • 한국잡초학회지
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    • 제12권4호
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    • pp.368-373
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    • 1992
  • 귀리에 alachlor를 처리(處理)할 때 변화(變化)되는 단백질(蛋白質)을 분석(分析)한 결과요약(結果要約)하면 다음과 같다. Alachlor 처리시(處理時) 단백질(蛋白質) 합성(合成)은 $1{\times}10^{-4}$M에서 5.8%, $1{\times}10^{-3}$M에서는 86.5% 억제(抑制)되는 현상(現象)을 보였지만 $1{\times}10^{-5}$M 이하(以下)의 농도(濃度)에서는 증가(增加)되는 현상(現象)을 보였다. 귀리의 근단분열조직(根端分裂組織)에서 추출(抽出)된 단백질(蛋白質)은 100 kd 이하(以下)의 polypeptide로 구성(構成)되어 있으며, alachlor 처리시(處理時) 47kd 이상(以上)의 고분자(高分子) 단백질(蛋白質)들은 억제(抑制)되었지만 23kd 이하(以下)의 저분자(低分子) 단백질(蛋白質)들은 오히려 증가(增加)되는 현상(現象)을 보였다. 이차원적(二次元的) 전기영동(電氣泳動) 결과(結果) alachlor 처리시(處理時) 83kd의 1, 2 spot, 70kd의 3, 4 spot, 47.5kd의 5, 6 spot의 polypeptide가 억제(抑制)된 반면(反面), 20kd의 7 spot, 16kd 의 8, 9, 10 spot는 증가(增加)되는 현상(現象)을 보였다. 또한 귀리 근단(根端) 분열조직(分裂組織)은 산성(酸性) 단백질(蛋白質)로서, 주(主)로 중성(中性) 부위(部位)에 큰 polypeptide spots들이 존재(存在)하고 있으며, 작은 spots을은 산성(酸性)쪽에 분리(分離)되어 나타났다.

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Immunological Characterization of Full and Truncated Recombinant Clones of ompH(D:4) Obtained from Pasteurella multocida (D:4) in Korea

  • Kim, Young-Hwan;Cheong, Ki-Young;Shin, Woo-Seok;Hong, Sung-Youl;Woo, Hee-Jong;Kwon, Moo-Sik
    • Journal of Microbiology and Biotechnology
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    • 제16권10호
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    • pp.1529-1536
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    • 2006
  • We cloned a gene of ompH(D:4) from pigs infected with P. multocida D:4 in Korea [16]. The gene is composed of 1,026 nucleotides coding 342 amino acids (aa) with a signal peptide of 20 aa (GenBank accession number AY603962). In this study, we analyzed the ability of the ompH(D:4) to induce protective immunity against a wild-type challenge in mice. To determine appropriate epitope(s) of the gene, one full and three different types of truncated genes of the ompH(D:4) were constructed by PCR using pET32a or pRSET B as vectors. They were named ompH(D:4)-F (1,026 bp [1-1026] encoding 342 aa), ompH(D:4)-t1 (693 bp [55-747] encoding 231 aa), ompH(D:4)-t2 (561 bp [187-747] encoding 187 aa), and ompH(D:4)-t3 (540 bp [487-1026] encoding 180 aa), respectively. The genes were successfully expressed in Escherichia coli BL21(DE3). Their gene products, polypeptides, OmpH(D:4)-F, -t1, -t2, and -t3, were purified individually using nickel-nitrilotriacetic acid (Ni-NTA) affinity column chromatography. Their $M_rs$ were determined to be 54.6, 29, 24, and 23.2 kDa, respectively, using SDS-PAGE. Antisera against the four kinds of polypeptides were generated in mice for protective immunity analyses. Some $50{\mu}g$ of the four kinds of polypeptides were individually provided intraperitoneally with mice (n=20) as immunogens. The titer of post-immunized antiserum revealed that it grew remarkably compared with pre-antiserum. The lethal dose of the wild-type pathogen was determined at $10{\mu}l$ of live P. multocida D:4 through direct intraperitoneal (IP) injection, into post-immune mice (n=5, three times). Some thirty days later, the lethal dose ($10{\mu}l$) of live pathogen was challenged into the immunized mouse groups [OmpH(D:4)-F, -t1, -t2, and -t3; n=20 each, two times] as well as positive and negative control groups. As compared within samples, the OmpH(D:4)-F-immunized groups showed lower immune ability than the OmpH(D:4)-t1, -t2, and -t3. The results show that the truncated-OmpH(D:4)-t1, -t2, and -t3 can be used for an effective vaccine candidate against swine atrophic rhinitis caused by pathogenic P. multocida (D:4) isolated in Korea.