• Title/Summary/Keyword: cell wall protein

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The protective effect of Prunella vulgaris ethanol extract against vascular inflammation in TNF-α-stimulated human aortic smooth muscle cells

  • Park, Sun Haeng;Koo, Hyun Jung;Sung, Yoon Young;Kim, Ho Kyoung
    • BMB Reports
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    • v.46 no.7
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    • pp.352-357
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    • 2013
  • Atherosclerosis, which manifests as acute coronary syndrome, stroke, and peripheral arterial diseases, is a chronic inflammatory disease of the arterial wall. Prunella vulgaris, a perennial herb with a worldwide distribution, has been used as a traditional medicine in inflammatory disease. Here, we investigated the effects of P. vulgaris ethanol extract on TNF-${\alpha}$-induced inflammatory responses in human aortic smooth muscle cells (HASMCs). We found that P. vulgaris ethanol extract inhibited adhesion of monocyte/macrophage-like THP-1 cells to activated HASMCs. It also decreased expression of intercellular adhesion molecule-1, vascular cell adhesion molecule-1, E-selectin and ROS, No production in TNF-${\alpha}$-induced HASMCs and reduced NF-${\kappa}B$ activation. Furthermore, P. vulgaris extract suppressed TNF-${\alpha}$-induced phosphorylation of p38 mitogen-activated protein kinase (MAPK) and extracellular signal-regulated kinase (ERK). These results demonstrate that P. vulgaris possesses anti-inflammatory properties and can regulate TNF-${\alpha}$-induced expression of adhesion molecules by inhibiting the p38 MAPK/ERK signaling pathway.

Protective Effect of Right Ventricular Mitochondrial Damage by Cyclosporine A in Monocrotaline-induced Pulmonary Hypertension

  • Lee, Dong Seok;Jung, Yong Wook
    • Korean Circulation Journal
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    • v.48 no.12
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    • pp.1135-1144
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    • 2018
  • Background and Objectives: Mitochondria play a key role in the pathophysiology of heart failure and mitochondrial permeability transition pore (MPTP) play a critical role in cell death and a critical target for cardioprotection. The aim of this study was to evaluate the protective effects of cyclosporine A (CsA), one of MPTP blockers, and morphological changes of mitochondria and MPTP related proteins in monocrotaline (MCT) induced pulmonary arterial hypertension (PAH). Methods: Eight weeks old Sprague-Dawley rats were randomized to control, MCT (60 mg/kg) and MCT plus CsA (10 mg/kg/day) treatment groups. Four weeks later, right ventricular hypertrophy (RVH) and morphological changes of right ventricle (RV) were done. Western blot and reverse transcription polymerase chain reaction (RT-PCR) for MPTP related protein were performed. Results: In electron microscopy, CsA treatment prevented MCT-induced mitochondrial disruption of RV. RVH was significantly increased in MCT group compared to that of the controls but RVH was more increased with CsA treatment. Thickened medial wall thickness of pulmonary arteriole in PAH was not changed after CsA treatment. In western blot, caspase-3 was significantly increased in MCT group, and was attenuated in CsA treatment. There were no significant differences in voltage-dependent anion channel, adenine nucleotide translocator 1 and cyclophilin D expression in western blot and RT-PCR between the 3 groups. Conclusions: CsA reduces MCT induced RV mitochondrial damage. Although, MPTP blocking does not reverse pulmonary pathology, it may reduce RV dysfunction in PAH. The results suggest that it could serve as an adjunctive therapy to PAH treatment.

Comparing Protein Expression in Erwinia amylovora Strain TS3128 Cultured under Three Sets of Environmental Conditions

  • Lee, Jongchan;Choi, Junhyeok;Lee, Jeongwook;Cho, Yongmin;Kang, In-Jeong;Han, Sang-Wook
    • The Plant Pathology Journal
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    • v.38 no.4
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    • pp.410-416
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    • 2022
  • Erwinia amylovora, the causal agent of fire-blight disease in apple and pear trees, was first isolated in South Korea in 2015. Although numerous studies, including omics analyses, have been conducted on other strains of E. amylovora, studies on South Korean isolates remain limited. In this study, we conducted a comparative proteomic analysis of the strain TS3128, cultured in three media representing different growth conditions. Proteins related to virulence, type III secretion system, and amylovoran production, were more abundant under minimal conditions than in rich conditions. Additionally, various proteins associated with energy production, carbohydrate metabolism, cell wall/membrane/envelope biogenesis, and ion uptake were identified under minimal conditions. The strain TS3128 expresses these proteins to survive in harsh environments. These findings contribute to understanding the cellular mechanisms driving its adaptations to different environmental conditions and provide proteome profiles as reference for future studies on the virulence and adaptation mechanisms of South Korean strains.

Proteomic and Phenotypic Analyses of a Putative YggS Family Pyridoxal Phosphate-Dependent Enzyme in Acidovorax citrulli

  • Lynn Heo;Yongmin Cho;Junhyeok Choi;Jeongwook Lee;Yoobin Han;Sang-Wook Han
    • The Plant Pathology Journal
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    • v.39 no.3
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    • pp.235-244
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    • 2023
  • Acidovorax citrulli (Ac) is a phytopathogenic bacterium that causes bacterial fruit blotch (BFB) in cucurbit crops, including watermelon. However, there are no effective methods to control this disease. YggS family pyridoxal phosphate-dependent enzyme acts as a coenzyme in all transamination reactions, but its function in Ac is poorly understood. Therefore, this study uses proteomic and phenotypic analyses to characterize the functions. The Ac strain lacking the YggS family pyridoxal phosphate-dependent enzyme, AcΔyppAc(EV), virulence was wholly eradicated in geminated seed inoculation and leaf infiltration. AcΔyppAc(EV) propagation was inhibited when exposed to L-homoserine but not pyridoxine. Wild-type and mutant growth were comparable in the liquid media but not in the solid media in the minimal condition. The comparative proteomic analysis revealed that YppAc is primarily involved in cell motility and wall/membrane/envelop biogenesis. In addition, AcΔyppAc(EV) reduced biofilm formation and twitching halo production, indicating that YppAc is involved in various cellular mechanisms and possesses pleiotropic effects. Therefore, this identified protein is a potential target for developing an efficient anti-virulence reagent to control BFB.

The Role of Nitric Oxide on the Growth Regulation of Chinese Cabbage (Brassica campestris L.) Primary Leaves (배추 (Brassica campestris L.) 제 1엽의 생장조절에 대한 Nitric Oxide의 역할)

  • Ham Jeong-Hun;Jin Chang-Duck
    • Journal of Plant Biotechnology
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    • v.32 no.4
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    • pp.293-300
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    • 2005
  • The possible role of nitric oxide (NO)-induced cell division was investigated to explain the physiologycal effects of a NO donor, sodium nitroprusside (SNP) on the growth of primary leaves in chinese cabbage seedling plants. Exogenous treatment of SNP to chinese cabbage plants for 8 days at different concentrations (0, 200, 500 and 1000 ${\mu}M$) affected the leaf growth in a concentration-dependent manner, showing a maximum growth at $200\;{\mu}M$. In accordance with leaf growth responses, the chlorophyll and soluble protein contents increased strongly to 142% and 134% of control at $200\;{\mu}M$ SNP, respectively. However, a very little decrease in chlorophyll and a 13%> decrease in protein were observed at $1000\;{\mu}M$ SNP. In addition, the content of DNA and RNA also increased maximumly to 142% and 139% of the control at $200\;{\mu}M$ SNP, respectively, whereas they decreased to 80% and 84% of the control at $1000\;{\mu}M$ SNP. With respect to the development of enzymes related to cell wall synthesis, $200\;{\mu}M$ SNP led to the maximum activities in both phenylalanine ammonia-lyase (212% of the control) and guaiacol peroxidase (134% of the control). However, the activities of both enzymes were not modified significantly at $1000\;{\mu}M$ SNP. In conclusion, these results suggest that the enhancement of leaf growth in chinese cabbage plants by SNP at the effective concentration was probably due to the NO ability in the induction of cell division.

The Use of Lupins in Feeding Systems - Review -

  • Petterson, D.S.
    • Asian-Australasian Journal of Animal Sciences
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    • v.13 no.6
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    • pp.861-882
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    • 2000
  • The seed, or grain, of modern cultivars of Lupinus angustifolius, commonly known as Australian sweet lupins (ASL), is an established feed resource for the intensive animal industries of Australia, Japan, Korea and several other countries in Asia and Europe. Since the introduction of ASL to the world marketplace about 25 years ago, researchers in many countries have found them to be a valuable component of the diet of beef and dairy cattle, sheep, pigs, poultry, finfish and crustaceans. The seed of ASL contains ~32% crude protein (CP) (~35% DM basis) and 5% oil. The main storage carbohydrates in the seed are the ${\beta}$-galactans that comprise most of the cell-wall material of the kernel and the cellulose and hemicellulose of the thick seed coats. ASL seeds contain about 40% non-starch polysaccharides (NSP) and a negligible amount of starch. This makes them an excellent ingredient for ruminant diets, as the risk of acidosis is very low. The seed of modern cultivars of domesticated Lupinus species contain negligible amounts of lectins and trypsin inhibitors so they do not require preheating before being used as an ingredient in feeds for monogastric species. They have a high digestibility coefficient for protein, >90% for most species, but a low energy digestibility, ~60%, which is mostly due to the high content of NSP. The low content of methionine (0.22%) and of lysine (1.46%) is typical of the legumes. The lysine availability for pigs is >70%. Lupin kernels contain ~39% CP (~42% DM basis), 6% oil and 30% NSP. They have a higher digestible energy for pigs and finfish and a higher metabolisable energy for poultry than whole seed. Commercial operations rarely achieve complete separation of kernel from hull and it is more likely that the kernel fraction, called splits or meats, will contain ~36% CP. The replacement of soybean meal or peas with ASL in cereal-based diets for most intensively reared animals, birds and fish is possible provided lysine, methionine and digestible energy levels are kept constant. This makes ASL economically competitive in many, but not all, circumstances.

Functional Characterization and Application of the HpOCH2 Gene, Encoding an Initiating $\alpha$l,6-Mannosyltransferase, for N-glycan Engineering in the Methylotrophic Yeast Hansenula polymorpha

  • Kim, Moo-Woong;Kim, Eun-Jung;Kim, Jeong-Yoon;Rhee, Sang-Ki;Kang, Hyun-Ah
    • Proceedings of the Korean Society for Applied Microbiology Conference
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    • 2004.06a
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    • pp.278-281
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    • 2004
  • The $\alpha$1,6-mannosyltransferase encoded by Saccharomyces cerevisiae OCH1 plays a key role for the outer chain initiation of the N-linked oligosaccharides. A search for Hansenula polymorpha genes homologous to S. cerevisiae OCHI (ScOCH1) has revealed seven open reading frames (ORF100, ORF142, ORF168, ORF288, ORF379, ORF576, ORF580). All of the seven ORFs are predicted to be a type II integral membrane protein containing a transmembrane domain near the amino-terminal region and has a DXD motif, which has been found in the active site of many glycosyltransferases. Among this seven-membered OCH1 gene family of H. polymorpha, we have carried out a functional analysis of H. polymorpha ORF168 (HpOCH2) showing the highest identity to ScOCH1. Inactivation of this protein by disruption of corresponding gene resulted in several phenotypes suggestive of cell wall defects, including hypersensitivity to hygromycin B and sodium deoxycholate. The structural analysis of N-glycans synthesized in HpOCH2-disrupted strain (Hpoch2Δ) and the in vitro $\alpha$1,6-mannosyltransferase activity assay strongly indicate that HpOch2p is a key enzyme adding the first $\alpha$1,6-mannose residue on the core glycan Man$_{8}$GlcNAc$_2$. The Hpoch2Δ was further genetically engineered to synthesize a recombinant glycoprotein with the human compatible N-linked oligosaccharide, Man$_{5}$GlcNAc$_2$, by overexpression of the Aspergillus saitoi $\alpha$1,2-mannosidase with the 'HDEL” ER retention signal.gnal.

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Tissue-dependent variation of protease expression phenotype in mouse peritoneal mast cells (마우스복강비만세포에서 프로테아제 발현 표현형의 조직 의존적 변화)

  • Lee, Young-Mi
    • Korean Journal of Veterinary Research
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    • v.41 no.4
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    • pp.543-548
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    • 2001
  • To examine the fate of the injected peritoneal mast cells (PMCs), we injected PMCs (500 or $10^5$) derived from WBB6F1-green fluorescent protein(GFP) mice into stomach wall of $WBB6F1-W/W^v$ mice. When 500 PMCs were injected, the proportion of alcian blue $(AB)^+$ mast cells to $GFP^+$ mast cells in the muscle was 25.0% on day 1, but decreased to 0.9% on day 7. Then, it increased to 98.2% on day 35. In contrast,$GFP^+$ mast cells in the mucosa were not detectable on day 1, 3, and 7 after injection. On day 35, the proportion of $AB^+$ mast cells to $GFP^+$ mast cells in the mucosa was 97.0%. When $10^5$ PMCs were injected, the proportion of $AB^+$ mast cells to $GFP^+$ mast cells in the muscle was more than 88.2%, and that in the mucosa was more than 86.3% from day 1 through 35 after injection. These results indicated that percentage of degranulation on day 1, 3, 7, 14 after injection of 500 PMCs was significantly higher than that after injection of $10^5$ PMCs. Futhermore, when 500 PMCs were injected, protease expression phenotypes of PMCs changed from day 14 after injection. When $10^5$ PMCs were injected, protease expression phenotype of PMCs did not change after injection. Such degranulated PMCs may acquire the new phenotype and adapt the new tissue.

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Isolation and Properties of a Protein, RCG-2, Having Chitinase, ${\beta}-1,3-Glucanase$ and Lysozyme Activities from Rice Leaves (Chitinase, ${\beta}-1,3-glucanase$ 및 lysozyme 효소활성을 보유한 벼잎 산성단백질 RCG-2)

  • Um, Sung-Yon;Kim, Su-Il
    • Applied Biological Chemistry
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    • v.37 no.1
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    • pp.49-55
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    • 1994
  • An acidic protein, RCG-2, containing chitinase and ${\beta}-1,3-glucanase$ activity conccurrently was purified from rice leaves by chromatofocusing and gel slicing. The purified enzyme gave a single band on polyacrylamide gel electrophoresis and its molecular weight was appeared to be 29.7 kd using SDS-PAGE. This enzyme also had lysozyme activity. The optimal temperature for both enzyme activities was $40^{\circ}C$, optimal pH were 4.0 for chitinase activity and 7.0 for ${\beta}-1,3-glucanase$ activity. $K_M$ and $V_{max}$ values for chitinase were 7.86 mM and $0.025\;{\mu}M/min.$, and those for ${\beta}-1,3-glucanase$ were 5.95 mM and $0.16\;{\mu}M/min.$ respectively. TLC analysis of the enzyme hydrolysates of chitooligosaccharides indicated that this enzyme acts as endochitinase.

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Cloning and Molecular Characterization of ${\beta}$-1,3-Glucan Synthase from Sparassis crispa

  • Yang, Yun Hui;Kang, Hyeon-Woo;Ro, Hyeon-Su
    • Mycobiology
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    • v.42 no.2
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    • pp.167-173
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    • 2014
  • A ${\beta}$-glucan synthase gene was isolated from the genomic DNA of polypore mushroom Sparassis crispa, which reportedly produces unusually high amount of soluble ${\beta}$-1,3-glucan (${\beta}$-glucan). Sequencing and subsequent open reading frame analysis of the isolated gene revealed that the gene (5,502 bp) consisted of 10 exons separated by nine introns. The predicted mRNA encoded a ${\beta}$-glucan synthase protein, consisting of 1,576 amino acid residues. Comparison of the predicted protein sequence with multiple fungal ${\beta}$-glucan synthases estimated that the isolated gene contained a complete N-terminus but was lacking approximately 70 amino acid residues in the C-terminus. Fungal ${\beta}$-glucan synthases are integral membrane proteins, containing the two catalytic and two transmembrane domains. The lacking C-terminal part of S. crispa ${\beta}$-glucan synthase was estimated to include catalytically insignificant transmembrane ${\alpha}$-helices and loops. Sequence analysis of 101 fungal ${\beta}$-glucan synthases, obtained from public databases, revealed that the ${\beta}$-glucan synthases with various fungal origins were categorized into corresponding fungal groups in the classification system. Interestingly, mushrooms belonging to the class Agaricomycetes were found to contain two distinct types (Type I and II) of ${\beta}$-glucan synthases with the type-specific sequence signatures in the loop regions. S. crispa ${\beta}$-glucan synthase in this study belonged to Type II family, meaning Type I ${\beta}$-glucan synthase is expected to be discovered in S. crispa. The high productivity of soluble ${\beta}$-glucan was not explained but detailed biochemical studies on the catalytic loop domain in the S. crispa ${\beta}$-glucan synthase will provide better explanations.