• Title, Summary, Keyword: Proteins

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Induction of Immune Responses by Two Recombinant Proteins of Brucella abortus, Outer Membrane Proteins 2b Porin and Cu/Zn Superoxide Dismutase, in Mouse Model

  • Sung, Kyung Yong;Jung, Myunghwan;Shin, Min-Kyoung;Park, Hyun-Eui;Lee, Jin Ju;Kim, Suk;Yoo, Han Sang
    • Journal of Microbiology and Biotechnology
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    • v.24 no.6
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    • pp.854-861
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    • 2014
  • The diagnosis of Brucella abortus is mainly based on serological methods using antibody against LPS, which has diagnostic problems. Therefore, to solve this problem, we evaluated two proteins of B. abortus, Cu/Zn superoxide dismutase (SodC) and outer membrane proteins 2b porin (Omp2b). The genes were cloned and expressed in a pMAL system, and the recombinant proteins, rOmp2b and rSodC, were purified as fusion forms with maltose-binding protein. The identity of the proteins was confirmed by SDS-PAGE and Western blot analysis with sera of mice infected with B. abortus. Production of cytokines and nitric oxide (NO) was investigated in RAW 264.7 cells and mouse splenocytes after stimulation with the proteins. Moreover, cellular and humoral immune responses were investigated in BALB/c mice after immunization with the proteins. TNF-${\alpha}$, IL-6, and NO were significantly inducible in RAW 264.7 cells. Splenocytes of naive mice produced IFN-${\gamma}$ and IL-4 significantly by stimulation. Moreover, number of IgG, IFN-${\gamma}$, and IL-4 producing cells were increased in immunized mice with the two proteins. Production of IgG and IgM with rOmp2b was higher than those with rSodC in immunized mice. These results suggest that the two recombinant proteins of B. abortus may be potential LPS-free proteins for diagnosis.

PEGYLATION: Novel Technology to Enhance Therapeutic Efficacy of Proteins and Peptides (PEG 접합: 단백질 및 펩타이드 치료제의 약효를 증가시키는 새로운 기술)

  • Park, Myung-Ok;Lee, Kang-Choon
    • Journal of Pharmaceutical Investigation
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    • v.30 no.2
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    • pp.73-83
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    • 2000
  • Polyethylene glycol (PEG) is a water soluble, biocompatible, non-toxic polymer and PEGylation is a well established technique for the modification of therapeutic proteins and peptides. PEG-protein drugs have been extensively studies in relation to therapies for various diseases: cancer, inflammation and others. The covalent attachment of PEG to proteins and peptides prolonged plasma half-life, reduced antigenicity and immunogenicity, increased thermal and mechanical stability, and prevented degradation by enzymes. Several chemical groups for general and site specific conjugation have been exploited to activate PEG for amino group, carboxyl group, and cysteine groups. PEGylation of many proteins and peptides have been studied to enhance their properties for the potential uses. Also, the different positional isomers in several PEG-proteins have shown the difference in vivo stability and biological indicating that the site of PEG molecule attachment is one of the important factor to develop PEG-proteins as potential therapeutic agents.

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The Effect of Animal Protein and Vegetable Protein Diet on Cholesterol Metabolism of Rats (동물성단백(動物性蛋白) 및 식물성단백(植物性蛋白)이 Cholesterol 대사(代謝)에 미치는 영향(影響))

  • Ahn, J.Y.
    • Journal of Nutrition and Health
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    • v.2 no.4
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    • pp.127-134
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    • 1969
  • Total and esterified cholesterol content was determined in the rat administered animal and vegetable proteins for 16 weeks. The cholesterol biosynthetic activity of the liver was also measured in these rats by the $acetate-C^{14}$ incorporation rate. The results obtained were as follows. (1) Serum total cholesterol content was increased by the administration of animal proteins and decreased by that of vegetable proteins. (2) Liver cholesterol content was increased by animal proteins and decreased by at of vegetable proteins. (3) Cholesterol biosynthetic activity of the liver was increased by the animal proteins and decreased by the vegetable proteins.

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Purification and Characterization of Crystalins by Aqueous Two-Phase Extraction

  • Bermudez, Ondrea;Forciniti, Daniel
    • Biotechnology and Bioprocess Engineering:BBE
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    • v.6 no.6
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    • pp.395-401
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    • 2001
  • Crystallins are a family of water-soluble proteins that constitute up to 90% of the wa-ter-soluble proteins in mammalian eye lenses, We present in this paper an alternative purification method for these proteins using polyethylene glycol/dextran aqueous two-phase extraction. Un-der the appropriate conditions, we were able to recover the γ-crystallin fraction essentially free of the remaining proteins. High concentrations of salt at a neutral pH maximize the recovery of γ-crystallins in the top phase and minimize the contamination by the other proteins present in the lenses. The proposed protocol decreases the separation time by about 50%. The complex partition behavior observed for these proteins reflects a delicate balance between protein/phase-forming species(various polymers and salts) and protein interactions. This is evidenced, in part, by the role played by the largest proteins in this group as a "pseudo"phase-forming species.

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Effects of proteins modified by enzymically oxidized caffic acid on yhe concentration of serum cholestrol of rats, part II (효소적 갈변 반응에 의하여 생성된 갈변 물질이 휜쥐 혈청콜레스테롤 농도에 미치는 영향)

  • 조영수;정순재
    • Journal of Life Science
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    • v.5 no.2
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    • pp.57-62
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    • 1995
  • Casein or soybean protein was subjected to there action with caffeic acidtyrosinase system at 30-35$\circ$C, pH 6.8 with aeration for 5hr. The resulting brown proteins were washed with acetone until the washings were on longer colored. However, modified protein still retained a light brown. The effects of the modified proteins and brown compounds on male Wistar strain rats were studied by pair-feeding of a cholesterol-free diet for 14days. Significant decrease in protein digestibility for the rats fed with the modified proteins were observed. Weight gain and protein digestibility were not influenced by feeding brown compounds, but the feeding of brown compound from casein caused an enlargement of caecum. The concentrations of serum cholesterol and triglyceride in the rats fed with modified proteins and brown compounds were mostly unchanged against the rats fed with untreated proteins. These results suggest that the decrease in protein digestibility induced by enzymic browning-reaction did not cause the decrease in concentration of serum cholesterol.

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Stability of Retroviral Vectors Against Ultracentrifugation Is Determined by the Viral Internal Core and Envelope Proteins Used for Pseudotyping

  • Kim, Soo-hyun;Lim, Kwang-il
    • Molecules and Cells
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    • v.40 no.5
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    • pp.339-345
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    • 2017
  • Retroviral and lentiviral vectors are mostly pseudotyped and often purified and concentrated via ultracentrifugation. In this study, we quantified and compared the stabilities of retroviral [murine leukemia virus (MLV)-based] and lentiviral [human immunodeficiency virus (HIV)-1-based] vectors pseudotyped with relatively mechanically stable envelope proteins, vesicular stomatitis virus glycoproteins (VSVGs), and the influenza virus WSN strain envelope proteins against ultracentrifugation. Lentiviral genomic and functional particles were more stable than the corresponding retroviral particles against ultracentrifugation when pseudotyped with VSVGs. However, both retroviral and lentiviral particles were unstable when pseudotyped with the influenza virus WSN strain envelope proteins. Therefore, the stabilities of pseudotyped retroviral and lentiviral vectors against ultracentrifugation process are a function of not only the type of envelope proteins, but also the type of viral internal core (MLV or HIV-1 core). In addition, the fraction of functional viral particles among genomic viral particles greatly varied at times during packaging, depending on the type of envelope proteins used for pseudotyping and the viral internal core.

Research on the Allergic Potential of Insecticidal CrylAc Proteins of Genetically Modified Rice

  • Son, Dae-Yeul
    • Food Science and Biotechnology
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    • v.15 no.3
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    • pp.385-391
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    • 2006
  • In Korea, different kinds of genetically modified (GM) crops are under development, including GM-rice expressing insecticidal crystal (Cry) proteins of Bacillus thuringiensis (Bt) modified to change a single amino acid. In this study, amino acid (aa) sequences of modified Cry proteins were compared to that of known allergens, and Cry proteins expressed in GM-rice were identified by using Cry protein specific polyclonal antibody. The antigen-antibody reactions were compared between GM and commercial rice to assess the allergic risk of Cry proteins. This analysis showed no known allergen to have more than 35% aa sequence homology with modified Cry proteins in Bt rice over an 80 aa window or to have more than 8 consecutive identical aa. Sera from allergic patients showed some IgE reactivity via immunoblotting and enzyme-linked immunosorbent assay (ELISA), although no differences were seen between GM and commercial rice. Based on these results we conclude that GM rice with modified Cry proteins has no differences in its protein composition or allergenicity relative to commercial rice.

Comparison of structure, function and regulation of plant cold shock domain proteins to bacterial and animal cold shock domain proteins

  • Chaikam, Vijay;Karlson, Dale T.
    • BMB Reports
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    • v.43 no.1
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    • pp.1-8
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    • 2010
  • The cold shock domain (CSD) is among the most ancient and well conserved nucleic acid binding domains from bacteria to higher animals and plants. The CSD facilitates binding to RNA, ssDNA and dsDNA and most functions attributed to cold shock domain proteins are mediated by this nucleic acid binding activity. In prokaryotes, cold shock domain proteins only contain a single CSD and are termed cold shock proteins (Csps). In animal model systems, various auxiliary domains are present in addition to the CSD and are commonly named Y-box proteins. Similar to animal CSPs, plant CSPs contain auxiliary C-terminal domains in addition to their N-terminal CSD. Cold shock domain proteins have been shown to play important roles in development and stress adaptation in wide variety of organisms. In this review, the structure, function and regulation of plant CSPs are compared and contrasted to the characteristics of bacterial and animal CSPs.

Amphetamine-induced ERM Proteins Phosphorylation Is through $PKC{\beta}$ Activation in PC12 Cells

  • Jeong, Ha-Jin;Kim, Jeong-Hoon;Jeon, Song-Hee
    • The Korean Journal of Physiology and Pharmacology
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    • v.15 no.4
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    • pp.245-249
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    • 2011
  • Amphetamine, a synthetic psychostimulant, is transported by the dopamine transporter (DAT) to the cytosol and increases the exchange of extracellular amphetamine by intracellular dopamine. Recently, we reported that the phosphorylation levels of ezrin-radixin-moesin (ERM) proteins are regulated by psychostimulant drugs in the nucleus accumbens, a brain area important for drug addiction. However, the significance of ERM proteins phosphorylation in response to drugs of abuse has not been fully investigated. In this study, using PC12 cells as an in vitro cell model, we showed that amphetamine increases ERM proteins phosphorylation and protein kinase C (PKC) ${\beta}$ inhibitor, but not extracellular signal-regulated kinase (ERK) or phosphatidylinositol 3-kinases (PI3K) inhibitors, abolished this effect. Further, we observed that DAT inhibitor suppressed amphetamine-induced ERM proteins phosphorylation in PC12 cells. These results suggest that $PKC{\beta}$-induced DAT regulation may be involved in amphetmaine-induced ERM proteins phosphorylation.

도축 폐혈액 단백질을 이용한 유산균체의 생산

  • 현창기;신현길
    • Microbiology and Biotechnology Letters
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    • v.25 no.2
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    • pp.218-223
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    • 1997
  • For the utilization of animal blood produced in slaughter for the cultivation of lactic acid bacteria, the nitrogen sources in a complex(MRS) medium were replaced by blood plasma proteins. Focusing the purpose on the industrial production of a probiotics, the hydrolytic activities of three industrially applicable proteases were compared for the effective digestion of the proteins, and Alcalase(the product of Novo Nordisk) was selected with comparatively high activity. The growth of Streptococcus thermophilus KCCM12020 was best among the four strains of lactic acid bacteria tested. With Alcalase-digested proteins in the medium, the growth rates and the final cell concentrations were higher than those with non-digested proteins. The cell mass produced in the medium containing blood proteins as nitrogen sources, $2.5{\times}10^9$ CFU/ml, was significantly high and about 70% of that in MRS medium, showing a great possibility for the utilization of animal blood proteins as economic nitrogen sources in the production of cell mass of lactic acid bacteria.

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