• Title/Summary/Keyword: Protein bodies

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Characteristics of Water-soluble Polysaccharide, Showing Inhibiting Activity on ${\alpha}-Glucosidase$, in Cordyceps militaris

  • Chung, Ha-Yull;Yoo, Mi-Kyong;Kawagishi, Hirokazu
    • Food Science and Biotechnology
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    • v.18 no.3
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    • pp.667-671
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    • 2009
  • Extract of water-soluble polysaccharide (CFWx), showing inhibiting activity on ${\alpha}-glucosidase$, was prepared from the fruiting bodies of Cordyceps militaris by hot-water extraction, and ethanol precipitation. Chemical characteristics of CFWx were as follows: carbohydrate content 30% including 16% of uronic acid; 51% protein content; monosaccharide composition, Man:Glu:Gal (30:43:27); molecular weight $3-5{\times}10^4$. CFWx was further purified by ion-exchange, gel-permeation, and affinity chromatography and $CFWx-AH-{\alpha}$ fraction was isolated. Fundamental structure of $CFWx-AH-{\alpha}$ was deduced as ${\alpha}-(1{\to}4$)-D-glucan with ${\alpha}-(1{\to}3$)- and/or ${\alpha}-(1{\to}6$)-D-glycosidic side chains based on methylation analysis.

Effect of Juvenile Hormone Analogue(JHA) on the Adult Development of the Silkworm, Bombyx mori (유약호르몬류 투여가 누에 성충화 발육에 미치는 영향)

  • Seong, Ju-Il;Im, Bong-Hak;Gang, Hyeon-A
    • Journal of Sericultural and Entomological Science
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    • v.35 no.1
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    • pp.28-35
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    • 1993
  • Treatment of juvenile hormone analogue(JHA) at the 5th instar larvae prolonged the duration of adult development one to one and half days as well as the elongation of feeding time with the increasing of larval body weight. Morphological observation and protein analysis in hemolymphs, integuements, alimentary canals, fat bodies and ovaries also revealed that the development of these tissues and organs for adultation are affected by the JHA treatment.

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Binding Characteristics to Mosquito-larval Midgut Proteins of the Cloned Domain II-III Fragment from the Bacillus thuringiensis Cry4Ba Toxin

  • Moonsom, Seangdeun;Chaisri, Urai;Kasinrerk, Watchara;Angsuthanasombat, Chanan
    • BMB Reports
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    • v.40 no.5
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    • pp.783-790
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    • 2007
  • Receptor binding plays an important role in determining host specificity of the Bacillus thuringiensis Cry $\delta$-endotoxins. Mutations in domains II and III have suggested the participation of certain residues in receptor recognition and insect specificity. In the present study, we expressed the cloned domain II-III fragment of Cry4Ba and examined its binding characteristics to mosquito-larval midgut proteins. The 43-kDa Cry4Ba-domain II-III protein over-expressed in Escherichia coli as inclusion bodies was only soluble when carbonate buffer, pH 10.0 was supplemented with 4M urea. After renaturation via stepwise dialysis and subsequent purification, the refolded domain II-III protein, which specifically reacts with anti Cry4Ba-domain III monoclonal antibody, predominantly exists as a $\beta$-sheet structure determined by circular dichroism spectroscopy. In vitro binding analysis to both histological midgut tissue sections and brush border membrane proteins prepared from susceptible Aedes aegypti mosquito-larvae revealed that the isolated Cry4Ba-domain II-III protein showed binding functionality comparable to the 65-kDa full-length active toxin. Altogether, the data present the 43-kDa Cry4Ba fragment comprising domains II and III that was produced in isolation was able to retain its receptor-binding characteristics to the target larval midgut proteins.

Soluble Expression of Recombinant Human Smp30 for Detecting Serum Smp30 Antibody Levels in Hepatocellular Carcinoma Patients

  • Zhang, Sheng-Chang;Huang, Peng;Zhao, Yong-Xiang;Liu, Shu-Yan;He, Shu-Jia;Xie, Xiao-Xun;Luo, Gou-Rong;Zhou, Su-Fang
    • Asian Pacific Journal of Cancer Prevention
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    • v.14 no.4
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    • pp.2383-2386
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    • 2013
  • Senescence marker protein 30 (SMP30), a hepatocellular carcinoma (HCC) associated antigen, was earlier shown by our research group to be highly expressed in HCC paracancerous tissues, but have low levels in HCC tissues. In order to detect anti-SMP30 antibody in serum of HCC patients, we established pET30a-SMP30 and pColdIII-SMP30 expression systems in Escherichia coli. However, the expression product was mainly in the form of inclusion bodies. In this research, we used several combinations of chaperones, four molecular chaperone plasmids with pET30a-SMP30 and five molecular chaperone plasmids with pColdIII-SMP30 to increase the amount of soluble protein. Results showed that co-expression of HIS-SMP30 with pTf16, combined with the addition of osmosis-regulator, and a two-step expression resulted in the highest enhancement of solubility. A total of 175 cases of HCC serum were studied by ELISA to detect anti-SMP30 antibody with recombinant SMP30 protein. Some 22 were positive and x2 two-sided tests all showed P>0.05, although it remained unclear whether there was a relationship between positive cases and clinical diagnostic data.

Characteristics of Cucumber mosaic virus Infecting Zucchini in Korea

  • Kim, Mi-Kyeong;Kwak, Hae-Ryun;Jeong, Seon-Gi;Ko, Sug-Ju;Lee, Su-Heon;Kim, Jeong-Soo;Kim, Kook-Hyung;Choi, Jang-Kyung;Choi, Hong-Soo;Cha, Byeong-Jin
    • The Plant Pathology Journal
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    • v.26 no.2
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    • pp.139-148
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    • 2010
  • A virus causing stunt, yellowing, severe mosaic, malformation symptoms on leaves and uneven development and malformation on fruits of zucchini was prevalent around Goseong, Gyeongsangnam-do, Korea. A survey conducted (2004) in the Goseong area revealed about 20% virus infection rate. The disease causative identified as Cucumber mosaic virus (CMV-Z1) was further characterized. The isolate induces mosaic symptoms on Cucumis sativus, while severe mosaic, stunt and malformation on C. pepo. Thin section analyses have shown that virus inclusions are formed in the cuticle layers as well as epidermal, parenchyma and collenchymas cells in virus-infected Nicotiana tabacum. CMV-Z1 isolate induced specific cytoplasmic inclusion bodies such as irregular clumps (IC), crystal (Cr) and irregular chloroplasts (ICh). IC was made up of virus particles interspersed with a darkly stained amorphous material and found both in the cytoplasm and vacuoles, whereas ICh and Cr were rarely found in the vacuoles. The genome of CMV-Z1 RNA-1 consists of 3359 nucleotide (nt) encoding 1a protein of 993 amino acids (aa). The CMV-Z1 RNA-2 was 3050 nt in length containing 2a (857 aa) and 2b (110 aa), while RNA-3 encoding 3a movement protein (279 aa) and coat protein (218 aa) was 2215 nt in length. Phylogenetic analyses of nucleotide sequences of CMV-Z1 isolate appeared it is more closely related to subgroup IA than to subgroup IB or II.

Changes of Texture, Soluble Solids and Protein during Cooking of Soybeans (콩의 조리과정 중 텍스쳐, 고형물 및 단백질의 변화)

  • Kim, Young-Ok;Jung, Hae-Ok;Rhee, Chong-Ouk
    • Korean Journal of Food Science and Technology
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    • v.22 no.2
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    • pp.192-198
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    • 1990
  • Texture, losses of total solids and proteins o) soybeans were studied during cooking at $100-135^{\circ}C$. The textural changes were measured using the puncture probe with an Instron Universal Testing Machine, and changes in microstructure of beans were observed with scanning electron microscopy during the cooking. The major effect observed was a breakdown of the cell walls and appearance of the protein bodies with soaking process. As the cooking time at $100^{\circ}C$ is longer, the separation of cells and changes in cell shape could be seen in the sample. The greater amounts of soluble solids were leached out with longer coo king time from the beans.

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Overexpression and Refolding of BACE2 (BACE2의 대량발현 및 리폴딩)

  • Park, Sun Joo;Tai, Shuaiqi;Lee, Yeon-Ji;Jeon, You-Jin;Kim, Yong-Tae
    • Korean Journal of Fisheries and Aquatic Sciences
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    • v.47 no.4
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    • pp.370-375
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    • 2014
  • BACE2 is a membrane-bound aspartic protease that is highly homologous with BACE1. While BACE1 processes the amyloid precursor protein (APP) at a key step in generating ${\beta}$-amyloid peptide and presumably causes Alzheimer's disease (AD), BACE2 has not been demonstrated to be involved in APP processing directly, and its physiological functions are unknown. To determine its function and to develop inhibitors from marine sources, we constructed an overexpression vector for producing BACE2. The gene encoding human BACE2 protease was amplified using the polymerase chain reaction and cloned into the pET11a expression vector, resulting in pET11a/BACE2. Recombinant BACE2 protease was overexpressed successfully in E. coli as inclusion bodies, refolded using the rapid-dilution method, and purified via two-step fast protein liquid chromatography using Sephacryl S-300 gel filtration and Resource-Q column chromatography. The BACE2 protease produced was an active form. This study provides an efficient method not only for studying the basic properties of BACE2, but also for developing inhibitors from natural marine sources.

Alzheimer's Disease and Apoptosis

  • Kim, Young-Hoon;Kim, Hye-Sun;Park, Cheol-Hyoung;Jeong, Sung-Jin;Kim, Young-Kyung;Kim, Sun-Hee;Lee, Sang-Kyeng;Suh, Yoo-Hun;Kim, Sung Su
    • Korean Journal of Biological Psychiatry
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    • v.5 no.1
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    • pp.66-70
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    • 1998
  • Apoptosis is a form of cell death in which the cells shrink and exhibit nuclear chromatin condensation and DNA fragmentation, and yet maintain membrane integrity. Many lines of evidence have shown that brain neurons are vulnerable to degeneration by apoptosis. Also it has been suggested that apoptosis is one of the mechanism contributing neuronal loss in Alzheimer's disease(AD), since the conditions in the disease($A{\beta}$ peptide, oxidative stress, low energy metabolism) are the inducers that activate apoptosis. Indeed some neurons in vulnerable regions of the AD brain show DNA damage, chromatin condensation, and apoptic bodies. Consistently, mutations in AD causative genes(Amyloid precursor protein, Presenilin-1 and Presenilin- 2) increase $A{\beta}$ $peptide_{1-42}(A{\beta}_{1-42})$ and sensitize neuronal cell to apoposis. However, several lines of evidence have shown that the location of neuronal loss and $A{\beta}$ peptide deposition is not correlated in AD brain and transgenic mice brain over-expressing $A{\beta}_{1-42}$. Taken together, these data may indicated that $A{\beta}$ peptide(and other causative factors of AD) can interact with other cellular insults or risk factors to exacerbate pathological mechansim of AD through apoptosis. Thus, this review discusses possible role and mechanism of apoptosis in AD.

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Distribution of Lipid and Lipase in Lipid-and Starch-Rich Seeds (지질 및 전분성 종자에서 지질 및 지질가수분해효소의 분포)

  • 김우갑
    • Journal of Plant Biology
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    • v.35 no.3
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    • pp.219-227
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    • 1992
  • Formation, cellular distribution and structural changes of storage lipid, and active site and cellular localization of lipase in endosperms and cotyledons of lipid-rich seeds such as Helianthus annuus, Ricinus communis and Pinus koraiensis, and in those of starch-rich seeds such as Pisum sativum and Zea mays were investigated in relation to the seed development by cytochemical methods. In endosperms and storage cotyledons of lipid- and starch-rich seeds after seed-gathering, there were widely distributed storage material which was composed of spherical protein bodies, spherosomes, and starch granules. But cellular organelles were hardly observed in the cytoplasm. Staining pattern of vesicles released from SER, and of low electron dense membraneous granules, which were perhaps at an early stage of spherosomes, were the same as in the spherosome. Electrondense granules released from RER were observed in the vicinity of plasma membrane. As a result of lipid staining, the spherosomes were more electron dense and were uniform as compared with the protein matrix within the protein body and cytoplasmic proteinaceous granules. The major component of the spherosome was determinated to be lipid. Spherosomes and vesicles containing SER-released materials showed the same as in the electron density. Lipase activity was especially strong in the inner region and on the surface of decomposed spherosomes and near the plasma membrane.mbrane.

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Ultrastructural changes of Endosperm Cells in Ginseng (Panax ginseng C.A. Meyer) Seeds during After-Ripening (인삼(Panax ginseng C.A. Meyer) 종자의 후숙에 따른 배유세포의 미세구조 변화)

  • 유성철
    • Journal of Plant Biology
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    • v.35 no.1
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    • pp.53-60
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    • 1992
  • This study has been carried out to investigate the ultrastructural changes in the associated with the disintegration of the storage materials in endosperm cell of ginseng (Panax ginseng C.A. Meyer) seed during after-ripening with light and electron microscope. The protein body of endosperm cells near the umbiliform layer showed various degenerative patterns, and so electron density of proteinaceous matrix was gradualJy decreased during afterripening. These results indicate that the decomposition of endosperm was already initiated during after-ripening. As the degeneration of endosperm was more progressed after the dehiscence of seed, non-decomposed part of protein body appeared amorphously with high electron density. Decomposed protein bodies were vacuolized with the loss of their matrix and gradually expanded by fusion. Also, spherosomes were gradually dissolved with the lowered electron density during the degeneration of endosperm. The vesicles of dictyosomes near the cell wall are observed in endosperm contacting with umbiliform layer and are fused with plasma membrane. Umbiliform layer which was the complex of the decomposed remnants of lysis and materials has strong stainability for toluidine blue and basic fuchsin.

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