• Title/Summary/Keyword: inducible proteins

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Anti-inflammatory Effects of Aster yomena Extracts by the Suppression of Inducible Nitric Oxide Synthase Expression

  • Kim, Ah-Yeon;Shin, Hyeon-Myeong;Kim, Ji-Soo;Shim, Hyun-Jin;Nam, Kung-Woo;Hwang, Kyung-A;Youn, Hyung-Sun
    • Biomedical Science Letters
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    • v.23 no.2
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    • pp.104-110
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    • 2017
  • Inflammation is a pathophysiological process that is known to be involved in numerous diseases. Microbial infection or tissue injury activates inflammatory responses, resulting in the induction of proinflammatory proteins including inducible nitric oxide synthase (iNOS). Aster yomena is used in traditional Korean remedies. Here, we investigated the effects of ethanol extracts of Aster yomena (EAY) on the expression of iNOS induced by ovalbumin (OVA), one of the major egg allergens, or lipopolysaccharide (LPS), a Toll-like receptor 4 agonist. EAY inhibited OVA- or LPS-induced $NF-{\kappa}B$ activation. EAY also suppressed OVA- or LPS-induced iNOS expression and nitrite production. These results suggest that EAY has the specific mechanism for anti-inflammatory responses and the potential to be developed as a potent anti-inflammatory and anti-allergic drug.

Functional Role of a Conserved Sequence Motif in the Oxygen-dependent Degradation Domain of Hypoxia-inducible Factor 1α in the Recognition of p53

  • Chi, Seung-Wook
    • Genomics & Informatics
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    • v.6 no.2
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    • pp.72-76
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    • 2008
  • Hypoxia-inducible factor $1{\alpha}\;(HIF1{\alpha})$ is a transcription factor that plays a key role in the adaptation of cells to low oxygen stress and oxygen homeostasis. The oxygen-dependent degradation (ODD) domain of $HIF1{\alpha}$ is responsible for the negative regulation of $HIF1{\alpha}$ in normoxia. The interactions of the $HIF1{\alpha}$ ODD domain with partner proteins such as von Hippel-Lindau tumor suppressor (pVHL) and p53 are mediated by two sequence motifs, the N- and C-terminal ODD(NODD and CODD). Multiple sequence alignment with $HIF1{\alpha}$ homologs from human, monkey, pig, rat, mouse, chicken, frog, and zebrafish has demonstrated that the NODD and CODD motifs have noticeably high conservation of the primary sequence across different species and isoforms. In this study, we carried out molecular dynamics simulation of the structure of the $HIF1{\alpha}$ CODD motif in complex with the p53 DNA-binding domain (DBD). The structure reveals specific functional roles of highly conserved residues in the CODD sequence motif of $HIF1{\alpha}$ for the recognition of p53.

Search for Novel Stress-responsive Protein Components Using a Yeast Mutant Lacking Two Cytosolic Hsp70 Genes, SSA1 and SSA2

  • Matsumoto, Rena;Rakwal, Randeep;Agrawal, Ganesh Kumar;Jung, Young-Ho;Jwa, Nam-Soo;Yonekura, Masami;Iwahashi, Hitoshi;Akama, Kuniko
    • Molecules and Cells
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    • v.21 no.3
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    • pp.381-388
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    • 2006
  • Heat shock proteins (Hsp) 70 are a ubiquitous family of molecular chaperones involved in many cellular processes. A yeast strain, ssa1/2, with two functionally redundant cytosolic Hsp70s (SSA1 and SSA2) deleted shows thermotolerance comparable to mildly heatshocked wild type yeast, as well as increased protein synthesis and ubiquitin-proteasome protein degradation. Since mRNA abundance does not always correlate well with protein expression levels it is essential to study proteins directly. We used a gel-based approach to identify stress-responsive proteins in the ssa1/2 mutant and identified 43 differentially expressed spots. These were trypsin-digested and analyzed by nano electrospray ionization liquid chromatography tandem mass spectrometry (nESI-LC-MS/MS). A total of 22 non-redundant proteins were identified, 11 of which were confirmed by N-terminal sequencing. Nine proteins, most of which were up-regulated (2-fold or more) in the ssa1/2 mutant, proved to be stress-inducible proteins such as molecular chaperones and anti-oxidant proteins, or proteins related to carbohydrate metabolism. Interestingly, a translational factor Hyp2p up-regulated in the mutant was also found to be highly phosphorylated. These results indicate that the cytosolic Hsp70s, Ssa1p and Ssa2p, regulate an abundance of proteins mainly involved in stress responses and protein synthesis.

Regulation of Class II Bacteriocin Production by Cell-Cell Signaling

  • Quadri, Luis E.N.
    • Journal of Microbiology
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    • v.41 no.3
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    • pp.175-182
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    • 2003
  • Production of ribosomally synthesized antimicrobial peptides usually referred to as bacteriocins is an inducible trait in several gram positive bacteria, particularly in those belonging to the group of lactic acid bacteria. In many of these organisms, production of bacteriocins is inducible and induction requires secretion and extracellular accumulation of peptides that act as chemical messengers and trigger bacteriocin production. These inducer peptides are often referred to as autoinducers and are believed to permit a quorum sensing-based regulation of bacteriocin production. Notably, the peptides acting as autoinducers are dedicated peptides with or without antimicrobial activity or the bacteriocins themselves. The autoinducer-dependent induction of bacteriocin production requires histidine protein kinases and response regulator proteins of two-component signal transduction systems. The current working model for the regulation of class II bacteriocin production in lactic acid bacteria and the most relevant direct and indirect pieces of evidence supporting the model are discussed in this minireview.

Differential Proteomic Analysis of Secreted Proteins from Cutinase-producing Bacillus sp. SB-007

  • Ban, Yeon-Hee;Jeon, Mi-Ri;Yoon, Ji-Hee;Park, Jae-Min;Um, Hyun-Ju;Kim, Dae-Soon;Jung, Seung-Ki;Kim, Keun-Young;Lee, Jee-Won;Min, Ji-Ho;Kim, Yang-Hoon
    • The Plant Pathology Journal
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    • v.24 no.2
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    • pp.191-201
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    • 2008
  • Bacillus sp. SB-007 was isolated from pea leaves harvested from the southwestern parts of South Korea through screening on a minimal medium containing 0.2% purified cutin for its ability to induce the cutinase production. However, no cutinase was produced when it was grown in a minimal medium containing 0.2% glucose. A proteomic approach was applied to separate and characterize these differentially secreted proteins. The expression level of 83 extracellular proteins of the cutinase-producing Bacillus sp. strain SB-007 incubated in a cutinase-induced medium increased significantly as compared with that cultured in a non cutinase-induced medium containing glucose. The extracellular proteome of Bacillus sp. SB-007 includes proteins from different functional classes, such as enzymes for the degradation of various macromolecules, proteins involved in energy metabolism, sporulation, transport/binding proteins and lipoproteins, stress inducible proteins, several cellular molecule biosynthetic pathways and catabolism, and some proteins with an as yet unknown function. In addition, the two protein spots showed little similarities with the known lipolytic enzymes in the database. These secreted proteome analysis results are expected to be useful in improving the Bacillus strains for the production of industrial cutinases.

Ovalbumin Induces Cyclooxygenase-2 and Inducible Nitric Oxide Synthase Expression (Ovalbumin에 의한 cyclooxygenase-2와 inducible nitric oxide synthase 유도)

  • Lee, A-Neum;Park, Se-Jeong;Jeong, Ae-Ri;Lee, Jae-Ran;Park, Hye-Jeong;Kim, Soo-Jung;Min, In-Soon;Youn, Hyung-Sun
    • Korean Journal of Food Science and Technology
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    • v.43 no.1
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    • pp.110-113
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    • 2011
  • Egg allergies are the most prevalent food hypersensitivity in children. The major egg allergens are proteins, such as ovalbumin (OVA) and ovomucoid (OVM), which are mainly contained in egg whites. OVA is the major protein in egg white, comprising 54% of the total protein content. OVA has been widely used in experimental inhalant and dietary allergy animal models, but its mechanism has not been clearly identified. In this study, we showed that OVA induced nuclear factor-${\kappa}B$ activation. OVA also induced the expression of cyclooxygenase-2 and inducible nitric oxide synthase. These data suggest new approaches for developing efficient anti-allergic strategies.

Loss of Specific Sequences in a Natural Variant of Potato Proteinase Inhibitor II Gene Results in a Loss of Wound-Inducible Gene Expression (감자의 단백질 분해효소 억제제 II 유전자의 특별한 염기서열의 자연적 제거로 인한 상처 유발성 발현의 소실)

  • Thornburg, Robert W.;Park, Sang-Gyu
    • Applied Biological Chemistry
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    • v.39 no.2
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    • pp.104-111
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    • 1996
  • We have isolated several proteinase inhibitor II genes pin2 from a Russet Burbank potato DNA library. One of these, pin2T was subcloned and a 1.8 kb Xbal/Nsil insert was sequenced. This fragment contained the complete Inhibitor II gene including 965 Up of flanking DNA upstream from the gene and 200 bp of flanking DNA downstream from the gene. The open reading frame encodes a protein that is similar to other reported proteinase Inhibitor II proteins. The DNA sequence of the 5' flanking region of pin2T from -714 to +1 is highly homologous (91% identity) with that of the previously isolated wound-inducible pin2K. There are, however, four small deletions in the pin2T promoter which are located at -221 to -200, -263 to -254, -523 to -426 and -759 to -708 relative to the transcription start site of the wound-inducible pin2K. Three of these deletions map to a portion of the promoter that controls the wound-inducibility of the proteinase inhibitor genes. Chimeric genes containing the promoter of the pin2T gene linked with the both CAT and GUS were constructed and transfered into tobacco plants. Analysis of these plants indicated that pin2T is not a wound-inducible gene but is expressed at low levels. Thus, wound-inducibility is lost with the concomitant natural deletion of three small regions of the promoter. Comparision of the sequences deleted in pin2T relative to the pin2K with Genebank sequences indicates that the deleted sequences contain a motif (consensus 5'-AGTAAA-3') that is found in many other wound-inducible genes but not easily found in the published promoter sequences of other plant genes. Nuclear proteins from unwounded and wounded potato leaves were bound to the proximal promoter region, downstream of the 5'-AGTAAA-3', of pin2T. The comparison of the pin2T gone with the pin2K gene indicates that the natural internal promoter deletions are likely responsible for loss of the wound-inducible phenotype in the pin2T gene.

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Variations in Protein Glycosylation in Hansenula polymorpha Depending on Cell Culture Stage

  • Kim, So-Young;Sohn, Jung-Hoon;Pyun, Yu-Ryang;Choi, Eui-Sung
    • Journal of Microbiology and Biotechnology
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    • v.17 no.12
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    • pp.1949-1954
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    • 2007
  • A simple way to prevent protein hyperglycosylation in Hansenula polymorpha was found. When glucose oxidase from Aspergillus niger and carboxymethyl cellulase from Bacillus subtilis were expressed under the control of an inducible methanol oxidase (MOX) promoter using methanol as a carbon source, hyperglycosylated forms occurred. In contrast, MOX-repressing carbon sources (e.g., glucose, sorbitol, and glycerol) greatly reduced the extent of hyperglycosylation. Carbon source starvation of the cells also reduced the level of glycosylation, which was reversed to hyperglycosylation by the resumption of cell growth. It was concluded that the proteins expressed under actively growing conditions are produced as hyperglycosylated forms, whereas those under slow or nongrowing conditions are as short-glycosylated forms. The prevention of hyperglycosylation in the Hansenula polymorpha expression system constitutes an additional advantage over the traditional Saccharomyces cerevisiae system in recombinant production of glycosylated proteins.

High-Level Production of Spider Silk Protein by Fed-Batch Cultivation of Recombinant Escherichia coli and Its Purification

  • Lee, Seok-Jae;Lee, Sang-Yeop
    • 한국생물공학회:학술대회논문집
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    • 2001.11a
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    • pp.719-722
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    • 2001
  • Silk proteins from Nephila clavipes are fibrous proteins containing repetitive sequences with both crystalline and amorphous domains. In order to obtain high-level production of silk protein, the synthetic genes had 16 contiguous units of the consensus repeat sequence of the silk protein were expressed in Escherichia coli BL21(DE3) under the strong inducible T7 promoter. For production of recombinant silk protein in large amounts, pH-stat fed-batch cultures were carried out. The recombinant silk protein was produced as soluble forms in E. coli, and the recombinant silk protein content was as high as 11% of the total protein. When cells were induced at $OD_{600}$ of 60, the amount of silk protein produced was 6.49 g/L. After simple purification steps, 9.2 mg of silk protein that was more than 80% pure was obtained from a 50 mL culture, and the recovery yield was 26.3%.

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