• Title/Summary/Keyword: major tail protein

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Spontaneous Release of Bacteriophage Particles by Lactobacillus rhamnosus Pen

  • Jarocki, Piotr;Podlesny, Marcin;Pawelec, Jaroslaw;Malinowska, Agata;Kowalczyk, Sylwia;Targonski, Zdzislaw
    • Journal of Microbiology and Biotechnology
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    • v.23 no.3
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    • pp.357-363
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    • 2013
  • The identification of bacteriophage proteins on the surface of Lactobacillus rhamnosus Pen was performed by LC-MS/MS analysis. Among the identified proteins, we found a phage-derived major tail protein, two major head proteins, a portal protein, and a host specificity protein. Electron microscopy of a cell surface extract revealed the presence of phage particles in the analyzed samples. The partial sequence of genes encoding the major tail protein for all tested L. rhamnosus strains was determined with specific primers designed in this study. Next, RT-PCR analysis allowed detection of the expression of the major tail protein gene in L. rhamnosus strain Pen at all stages of bacterial growth. The transcription of genes encoding the major tail protein was also proved for other L. rhamnosus strains used in this study. The present work demonstrates the spontanous release of prophage-encoded particles by a commercial probiotic L. rhamnosus strain, which did not significantly affect the bacterial growth of the analyzed strain.

In Vivo Protein Transduction: Delivery of PEP-1-SOD1 Fusion Protein into Myocardium Efficiently Protects against Ischemic Insult

  • Zhang, You-en;Wang, Jia-ning;Tang, Jun-ming;Guo, Ling-yun;Yang, Jian-ye;Huang, Yong-zhang;Tan, Yan;Fu, Shou-zhi;Kong, Xia;Zheng, Fei
    • Molecules and Cells
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    • v.27 no.2
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    • pp.159-166
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    • 2009
  • Myocardial ischemia-reperfusion injury is a medical problem occurring as damage to the myocardium following blood flow restoration after a critical period of coronary occlusion. Oxygen free radicals (OFR) are implicated in reperfusion injury after myocardial ischemia. The antioxidant enzyme, Cu, Zn-superoxide dismutase (Cu, Zn-SOD, also called SOD1) is one of the major means by which cells counteract the deleterious effects of OFR after ischemia. Recently, we reported that a PEP-1-SOD1 fusion protein was efficiently delivered into cultured cells and isolated rat hearts with ischemia-reperfusion injury. In the present study, we investigated the protective effects of the PEP-1-SOD1 fusion protein after ischemic insult. Immunofluorescecnce analysis revealed that the expressed and purified PEP-1-SOD1 fusion protein injected into rat tail veins was efficiently transduced into the myocardium with its native protein structure intact. When injected into Sprague-Dawley rat tail veins, the PEP-1-SOD1 fusion protein significantly attenuated myocardial ischemia-reperfusion damage; characterized by improving cardiac function of the left ventricle, decreasing infarct size, reducing the level of malondialdehyde (MDA), decreasing the release of creatine kinase (CK) and lactate dehydrogenase (LDH), and relieving cardiomyocyte apoptosis. These results suggest that the biologically active intact forms of PEP-1-SOD1 fusion protein will provide an efficient strategy for therapeutic delivery in various diseases related to SOD1 or to OFR.

Factors Influencing on the Drop of in vitro Protein Digestibility in Dried Fish Meat (건어육(乾魚肉) 저장(貯藏)중의 단백질(蛋白質) 소화율(消化率) 저하요인(低下要因))

  • Kim, Sang-Ae;Lee, Kang-Ho;Ryu, Hong-Soo
    • Journal of the Korean Society of Food Science and Nutrition
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    • v.15 no.1
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    • pp.45-55
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    • 1986
  • This paper aims to study the reactions of lipid or oxidized lipid with protein during drying and storing hair tail fish(Trichurus lepturus) and flounder(Kanakius kitaharai) being generally consumed as dried seafood products in Korea and their influence on the drop of in vitro protein digestibility of these fish meat. The results of the study are as follows: The digestibility of the raw materials of flounder and hair tail fish was 87.63% and 86.08% respectively, and that of sundried and hot air dried materials went down $1{\sim}2$ percent with drying process. But in case of defatted and sundried materials, the rate increased 85.15% and 87.15% respectivley. After 30 days of storage, the digestibility decreased in all materials, and hot air dried meat showed a significant decrease. Trypsin indigestible substrate (TIS) contents of flounder and hair tail fish, in case of raw materials were 0.88 and 0.96mg/g. solid repectiveiy and in case of defatted and sundried materials, TIS contents showed a low increase and digestibility showed a high increase. Brown pigment formation had a wide range of increase in case of the sundried and hot air dried materials and it was increased with duration of storage and temperature. The major fatty acids in the fats of hair tail fish and flounder were $C_{18:1},\;C_{16:0},\;C_{22:6}\;and\;C_{16:1}$ and rate of unsaturated to saturated fatty acids was 79.2:20.8 for flounder, 67.8:32.2 for hair tail fish. After 30 days of storage at room temperature. saturated fatty acids increased compared with the raw materials while unsaturated fatty acids showed a tendency to decrease. Avaialble lysine of hair tail fish was higher than that of flounder and both of them lost about 8.23% of that in raw materials after 30 days of storage.

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A Newly Isolated Bacteriophage, PBES 02, Infecting Cronobacter sakazakii

  • Lee, Hyung Ju;Kim, Wan Il;Kwon, Young Chan;Cha, Kyung Eun;Kim, Minjin;Myung, Heejoon
    • Journal of Microbiology and Biotechnology
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    • v.26 no.9
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    • pp.1629-1635
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    • 2016
  • A novel bacteriophage, PBES 02, infecting Cronobacter sakazakii was isolated and characterized. It has a spherical head of 90 nm in diameter and a tail of 130 nm in length, and belongs to Myoviridae as observed under a transmission electron microscope. The major virion protein appears to be 38 kilodaltons (kDa) in size. The latent period of PBES 02 is 30 min and the burst size is 250. Infectivity of the phage remained intact after exposure to temperatures ranging from 4℃ to 55℃ for 1 h. It was also stable after exposure to pHs ranging from 6 to 10 for 1 h. The phage effectively removed contaminating Cronobacter sakazakii from broth infant formula. PBES 02 has a double-stranded DNA genome of 149,732 bases. Its GC ratio is 50.7%. Sequence analysis revealed that PBES 02 has 299 open reading frames (ORFs) and 14 tRNA genes. Thirty-nine ORFs were annotated, including 24 related to replication and regulation functions, 10 related to structural proteins, and 5 related to DNA packaging. The genome of PBES 02 is closely related to that of two other C. sakazakii phages, CR3 and CR8. Comparison of DNA sequences of genes encoding the major capsid protein revealed a wide geographical distribution of related phages over Asia, Europe, and America.

Antidepressant effects of ginsenoside Rf on behavioral change in the glial degeneration model of depression by reversing glial loss

  • Kim, Yunna;Lee, Hwa-Young;Choi, Yu-Jin;Cho, Seung-Hun
    • Journal of Ginseng Research
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    • v.44 no.4
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    • pp.603-610
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    • 2020
  • Background: Depression is a common neuropsychiatric disease that shows astrocyte pathology. Ginsenoside Rf (G-Rf) is a saponin found in Panax ginseng which has been used to treat neuropsychiatric diseases. We aimed to investigate antidepressant properties of G-Rf when introduced into the L-alphaaminoadipic acid (L-AAA)-infused mice model which is representative of a major depressive disorder that features diminished astrocytes in the brain. Methods: L-AAA was infused into the prefrontal cortex (PFC) of mice to induce decrease of astrocytes. Mice were orally administered G-Rf (20 mg/kg) as well as vehicle only or imipramine (20 mg/kg) as controls. Depression-like behavior of mice was evaluated using forced swimming test (FST) and tail suspension test (TST). We observed recovery of astroglial impairment and increased proliferative cells in the PFC and its accompanied change in the hippocampus by Western blot and immunohistochemistry to assess the effect of G-Rf. Results: After injection of L-AAA into the PFC, mice showed increased immobility time in FST and TST and loss of astrocytes without significant neuronal change in the PFC. G-Rf-treated mice displayed significantly more decreased immobility time in FST and TST than did vehicle-treated mice, and their immobility time almost recovered to those of the sham mice and imipramine-treated mice. G-Rf upregulated glial fibrillary acidic protein (GFAP) expression and Ki-67 expression in the PFC reduced by L-AAA and also alleviated astroglial change in the hippocampus. Conclusion: G-Rf markedly reversed depression-like behavioral changes and exhibited protective effect against the astrocyte ablation in the PFC induced by L-AAA. These protective properties suggest that G-Rf might be a therapeutic agent for major depressive disorders.

Interaction of GAT1 with Ubiquitin-Specific Protease Usp14 in Synaptic Terminal (GAT1과 ubiquitin-specific protease Usp14의 결합)

  • Seog, Dae-Hyun;Kim, Sang-Jin;Joung, Young-Ju;Yea, Sung-Su;Park, Yeong-Hong;Kim, Moo-Seong;Moon, Il-Soo;Jang, Won-Hee
    • Journal of Life Science
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    • v.20 no.7
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    • pp.1005-1011
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    • 2010
  • $\gamma$-aminobutyric acid (GABA) is the major inhibitory neurotransmitter in the central nervous system. GABA transporters (GATs) control extracellular GABA levels by reuptake of released GABA from the synaptic cleft. However, how GATs are regulated has not yet been elucidated. Here, we used the yeast two-hybrid system to identify the specific binding protein(s) that interacts with the carboxyl (C)-terminal region of GAT1, the major isoform in the brain and find a specific interaction with the ubiquitin-specific protease 14 (Usp14), a deubiquitinating enzyme. Usp14 protein bound to the tail region of GAT1 and GAT2 but not to other GAT members in the yeast two-hybrid assay. The C-terminal region of Usp14 is essential for interaction with GAT1. In addition, these proteins showed specific interactions in the glutathione S-transferase (GST) pull-down assay. An antibody to GAT1 specifically co-immunoprecipitated Usp14 from mouse brain extracts. These results suggest that Usp14 may regulate the number of GAT1 at the cell surface.

First Isolation and Molecular Characterization of Bacteriophages Infecting Acidovorax citrulli, the Causal Agent of Bacterial Fruit Blotch

  • Rahimi-Midani, Aryan;Lee, Yong Seok;Kang, Se-Won;Kim, Mi-Kyeong;Choi, Tae-Jin
    • The Plant Pathology Journal
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    • v.34 no.1
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    • pp.59-64
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    • 2018
  • Bacteriophages of Acidovorax citrulli, the causal agent of bacterial fruit blotch, were isolated from 39 watermelon, pumpkin, and cucumber leaf samples collected from various regions of Korea and tested against 18 A. citrulli strains. Among the six phages isolated, ACP17 forms the largest plaque, and exhibits the morphology of phages in the Myoviridae family with a head diameter of $100{\pm}5nm$ and tail length of $150{\pm}5nm$. ACP17 has eclipse and latent periods of $25{\pm}5min$ and $50{\pm}5min$, respectively, and a burst size of 120. The genome of ACP17 is 156,281 base pairs with a G + C content of 58.7%, 263 open reading frames, and 4 transfer RNA genes. Blast search and phylogenetic analysis of the major capsid protein showed that ACP17 has limited homology to two Stentrophomonas phages, suggesting that ACP17 is a new type of Myoviridae isolated from A. citrulli.

Isolation and Characterization of a Bacteriophage Preying an Antifungal Bacterium

  • Rahimi-Midani, Aryan;Kim, Kyoung-Ho;Lee, Seon-Woo;Jung, Sang Bong;Choi, Tae-Jin
    • The Plant Pathology Journal
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    • v.32 no.6
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    • pp.584-588
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    • 2016
  • Several Bacillus species were isolated from rice field soils, and 16S rRNA gene sequence analysis showed that Bacillus cereus was the most abundant. A strain named BC1 showed antifungal activity against Rhizoctonia solani. Bacteriophages infecting strain BC1 were isolated from the same soil sample. The isolated phage PK16 had an icosahedral head of $100{\pm}5nm$ and tail of $200{\pm}5nm$, indicating that it belonged to the family Myoviridae. Analysis of the complete linear dsDNA genome revealed a 158,127-bp genome with G + C content of 39.9% comprising 235 open reading frames as well as 19 tRNA genes (including 1 pseudogene). Blastp analysis showed that the proteins encoded by the PK16 genome had the closest hits to proteins of seven different bacteriophages. A neighbor-joining phylogenetic tree based on the major capsid protein showed a robust clustering of phage PK16 with phage JBP901 and BCP8-2 isolated from Korean fermented food.