• Title/Summary/Keyword: accessory protein

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Functional Characterization and Proteomic Analysis of Porcine Deltacoronavirus Accessory Protein NS7

  • Choi, Subin;Lee, Changhee
    • Journal of Microbiology and Biotechnology
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    • v.29 no.11
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    • pp.1817-1829
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    • 2019
  • Porcine deltacoronavirus (PDCoV) is an emerging swine enteric coronavirus that causes diarrhea in neonatal piglets. Like other coronaviruses, PDCoV encodes at least three accessory or species-specific proteins; however, the biological roles of these proteins in PDCoV replication remain undetermined. As a first step toward understanding the biology of the PDCoV accessory proteins, we established a stable porcine cell line constitutively expressing the PDCoV NS7 protein in order to investigate the functional characteristics of NS7 for viral replication. Confocal microscopy and subcellular fractionation revealed that the NS7 protein was extensively distributed in the mitochondria. Proteomic analysis was then conducted to assess the expression dynamics of the host proteins in the PDCoV NS7-expressing cells. High-resolution two-dimensional gel electrophoresis initially identified 48 protein spots which were differentially expressed in the presence of NS7. Seven of these spots, including two up-regulated and five down-regulated protein spots, showed statistically significant alterations, and were selected for subsequent protein identification. The affected cellular proteins identified in this study were classified into functional groups involved in various cellular processes such as cytoskeleton networks and cell communication, metabolism, and protein biosynthesis. A substantial down-regulation of α-actinin-4 was confirmed in NS7-expressing and PDCoV-infected cells. These proteomic data will provide insights into the understanding of specific cellular responses to the accessory protein during PDCoV infection.

Expession of the Recombinant Klebsiella aerognes UreF Protein as a MalE Fusion

  • Kim, Keun-Young;Yang, Chae-Ha;Lee, Mann-Hyung
    • Archives of Pharmacal Research
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    • v.22 no.3
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    • pp.274-278
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    • 1999
  • Expression of the active urease of the enterobacterium, Klebsiella aerogens, requires the presence of the accessory genes (ureD, ureE, ureF, and ureG) in addition to the three structural genes (ureA, ureB, and ureC). These accessory genes are involved in functional assembly of the nickel-metallocenter for the enzyme. Characterization of ureF gene has been hindered, however, since the UreF protein is produced in only minute amount compared to other urease gene products. In order to overexpress the ureF gene, a recombinant pMAL-UreF plasmid was constructed from which the UreF was produced as a fusion with maltose-binding protein. The MBP-UreF fusion protein was purified by using an amylose-affinity column chromatography followed by an anion exchange column chromatography. Polyclonal antibodies raised against the fusion protein were purified and shown to specifically recognize both MBP and UreF peptides. The UreF protein was shown to be unstable when separated from MBP by digestion with factor Xa.

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Deletion Mutageneses of the Helicobacter pylori Urease Accessory Genes

  • Lee, Mann-Hyong;Sung, Jae-Young
    • Journal of Life Science
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    • v.9 no.1
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    • pp.5-8
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    • 1999
  • Helicobacter pylori is the etiologic agent of human gastritis and peptic ulceration and produces urease as the major protein component on its surface. H. pylori urease is known to serve as a major virulence factor and a potent immunogen. Deletion mutageneses were performed in the H. pylori urease accessory genes by using combinations of restriction enzymes and other DNA modifying enzymes in order to assess the function of these accessory gene products in the expression of the active urease. Selective disruptions in the accessory gene regions resulted in complete abolishment of the urease activity, which is consistent with other bacterial ureases. Interestingly, deletions in ureE-containing regions caused reduced expression of the structural enzyme subunits.

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Effects of Environmental Factors Such as Temperature and Salinity on Expression of Interleukin-1 Receptor Accessory Protein in the Red Seabream (Pagrus major) (온도 및 염분 등의 환경요인이 참돔(Pagrus major)의 Interleukin-1 Receptor Accessory Protein 발현에 미치는 영향)

  • Kang, Han Seung;Min, Byung Hwa
    • Journal of Marine Life Science
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    • v.2 no.2
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    • pp.70-74
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    • 2017
  • Interleukin-1 (IL-1) is one of the proinflammatory cytokines, after IL-1 binds to IL-1RI, IL-1RacP (interleukin-1 receptor accessory protein) joins with IL-1/IL-1RI to form a complex, and leading to cell activation. IL-1RAcP is involved in immune response, stress and apoptosis. The purpose of this study was to investigate the gene expression of IL-1RAcP in red seabream (Pagrus major) exposure to low water temperature (8℃, 33 psu) and low salinity (20℃, 10 psu). Results showed that, the expression of IL-1RAcP was significantly increased in the experiment groups, such as low water temperature (8℃, 33 psu), and low salinity (20℃, 10 psu). These results suggest that IL-1RAcP was played roles in biomarker gene on the environmental stress such as low water temperature and low salinity.

Constitutive Exocytosis of Seminal Fluid Proteins in Male Accessory Glands and Ejaculatory Duct of Drosophila melanogaster

  • Jo, Gyeong-Sang;Kim, Seong-Yun;Im, Jeom-Hui;Jeong, Gi-Hwa;Lee, Cheong-Ju
    • Animal cells and systems
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    • v.1 no.3
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    • pp.515-520
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    • 1997
  • Male accessory glands and ejaculatory duct of Drosophila melanogaster are reproductive organs which synthesize secretory seminal proteins. Several products of these organs involved in egg laying, receptivity, and sperm stability or storage were isolated from their lumens. Despite their secretory process play an important role, exocytosis pathway in these organs is not well known. In the present study, we characterized secretory protein profiles and determined their secretory mechanisms. Eight accessory gland secretory proteins and two ejaculatory duct secretory proteins were detected in their lumens. All these proteins were constitutively synthesized in these organs and secreted to their lumens. Secretion of newly synthesized proteins initiated at about 1 h after synthesis, and reached the peak at 4 h after synthesis. It seems that secretion of the proteins may occur via constitutive exocytosis pathway.

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Cytochemical and Immunocytochemical Study on the Cellulase Activity in the Accessory Glands of the Digestive System of the Oriental Land Snail, Nesiohelix

  • Jeong, Kye-Heon;Lee, Young-Seok;Shim, Yun-Bo
    • The Korean Journal of Malacology
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    • v.15 no.2
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    • pp.81-92
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    • 1999
  • The histochemical, cytochemical, and immunocytochemical investigations were conducted to find out the cellulase activity in the accessory glands of the digestive system of the oriental land snail Nesiohelix samarangae under the LM, SEM, and TEM. The cellulase activity was shown in the epithelium of th digestive gland by labelling with the immunogold (protein-A gold) particles. The epithelial cells showing the cellulase activity were Type 1 and Type 3 cells out of five types of the epithelial cells of the digestive gland. None of epithelial cells of the mucus gland and the salivary gland and the salivary gland were not labeled with the immunogold particles.

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