• Title/Summary/Keyword: Functional characterization

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Characterization of Functional Domains in NME1L Regulation of NF-κB Signaling

  • You, Dong-Joo;Park, Cho Rong;Mander, Sunam;Ahn, Curie;Seong, Jae Young;Hwang, Jong-Ik
    • Molecules and Cells
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    • v.39 no.5
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    • pp.403-409
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    • 2016
  • NME1 is a well-known metastasis suppressor which has been reported to be downregulated in some highly aggressive cancer cells. Although most studies have focused on NME1, the NME1 gene also encodes the protein (NME1L) containing N-terminal 25 extra amino acids by alternative splicing. According to previous studies, NME1L has potent anti-metastatic activity, in comparison with NME1, by interacting with $IKK{\beta}$ and regulating its activity. In the present study, we tried to define the role of the N-terminal 25 amino acids of NME1L in $NF-{\kappa}B$ activation signaling. Unfortunately, the sequence itself did not interact with $IKK{\beta}$, suggesting that it may be not enough to constitute the functional structure. Further construction of NME1L fragments and biochemical analysis revealed that N-terminal 84 residues constitute minimal structure for homodimerization, $IKK{\beta}$ interaction and regulation of $NF-{\kappa}B$ signaling. The inhibitory effect of the fragment on cancer cell migration and $NF-{\kappa}B$-stimulated gene expression was equivalent to that of whole NME1L. The data suggest that the N-terminal 84 residues may be a core region for the anti-metastatic activity of NME1L. Based on this result, further structural analysis of the binding between NME1L and $IKK{\beta}$ may help in understanding the anti-metastatic activity of NME1L and provide direction to NME1L and $IKK{\beta}$-related anti-cancer drug design.

Stage and Tissue Specific Expression of Four TCR Subunits in Olive Flounder (Paralichthys olivaceus)

  • Lee, Young Mee;Lee, Jeong-Ho;Noh, Jae Koo;Kim, Hyun Chul;Park, Choul-Ji;Park, Jong-Won;Hwang, In Joon;Kim, Sung Yeon
    • Development and Reproduction
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    • v.17 no.4
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    • pp.329-335
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    • 2013
  • TCR subunits are members of membrane-bound receptors which allow the fast and efficient elimination of the specific fish pathogens have regulated function in adaptive immunity. Sequence structure of TCR subunits have been reported for various teleosts, but the information of each TCR subunit functional characterization through expression analysis in fish was unknown. In this study, we examined the gene expression of TCR subunits in the early developmental stages and observed transcript levels in various tissues from healthy adult olive flounder by RT-PCR. The mRNA expression of alpha subunit was already detected in the previous hatching step. But the transcripts of another TCR subunit were not observed during embryo development and increased after hatching and maintained until metamorphosis at the same level. It was found that all TCR subunits mRNAs are commonly expressed in the immune-related organ such as spleen, kidney and gill, also weak expressed in fin and eye. TCR alpha and beta subunit were expressed in brain, whereas gamma and delta were not expressed same tissue. The sequence alignment analysis shows that there are more than 80% sequence homology between TCR subunits. Because it has a high similarity of amino acid sequence to expect similar in function, but expression analysis show that will have may functional diversity due to different time and place of expression.

Proteomic Functional Characterization of Bovine Stromal Vascular Cells from Omental, Subcutaneous and Intramuscular Adipose Depots

  • Rajesh, Ramanna Valmiki;Kim, Seong-Kon;Park, Mi-Rim;Nam, Jin-Seon;Kim, Nam-Kuk;Kwon, Seulemina;Yoon, Du-Hak;Kim, Tae-Hun;Lee, Hyun-Jeong
    • Asian-Australasian Journal of Animal Sciences
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    • v.24 no.1
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    • pp.110-124
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    • 2011
  • Anatomically separate fat depots differ in size, function, and contribution to pathological states such as the metabolic syndrome. We isolated pre-adipocytes from different adipose depots, omental, subcutaneous and intramuscular, of beef cattle, and cultured in vitro to determine the basis for the variations and attribute these variations to the inherent properties of adipocyte progenitors. The proliferating cells from all depots before the confluence were harvested and the proteome was analyzed by a functional proteomic approach, involving 2-DE and MALDI-TOF/TOF. More than 252 protein spots were identified, selected and analyzed by Image Master (ver 7.0) and MALDI-TOF/TOF. Further, our analysis showed that there were specific differences in proteome expression patterns among proliferating precursor cells from the three depots. Sixteen proteins were found to be differentially expressed and these were identified as proteins involved in cellular processes, heat shock/chaperones, redox proteins, cytoskeletal proteins and metabolic enzymes. The results also enabled us to understand the basic roles of these proteins in different inherent properties exhibited by adipose tissue depots.

Comparative Reverse Screening Approach to Identify Potential Anti-neoplastic Targets of Saffron Functional Components and Binding Mode

  • Bhattacharjee, Biplab;Vijayasarathy, Sandhya;Karunakar, Prashantha;Chatterjee, Jhinuk
    • Asian Pacific Journal of Cancer Prevention
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    • v.13 no.11
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    • pp.5605-5611
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    • 2012
  • Background: In the last two decades, pioneering research on anti-tumour activity of saffron has shed light on the role of crocetin, picrocrocin and safranal, as broad spectrum anti-neoplastic agents. However, the exact mechanisms have yet to be elucidated. Identification and characterization of the targets of bioactive constituents will play an imperative role in demystifying the complex anti-neoplastic machinery. Methods: In the quest of potential target identification, a dual virtual screening approach utilizing two inverse screening systems, one predicated on idTarget and the other on PharmMapper was here employed. A set of target proteins associated with multiple forms of cancer and ranked by Fit Score and Binding energy were obtained from the two independent inverse screening platforms. The validity of the results was checked by meticulously analyzing the post-docking binding pose of the picrocrocin with Hsp90 alpha in AutoDock. Results: The docking pose reveals that electrostatic and hydrogen bonds play the key role in inter-molecular interactions in ligand binding. Picrocrocin binds to the Hsp90 alpha with a definite orientation appropriate for nucleophilic attacks by several electrical residues inside the Hsp90-alpha ATPase catalytic site. Conclusion: This study reveals functional information about the anti-tumor mechanism of saffron bioactive constituents. Also, a tractable set of anti-neoplastic targets for saffron has been generated in this study which can be further authenticated by in vivo and in vitro experiments.

Synthesis and Photopolymerization Characterization of Propenyl Ether Monomers (프로페닐 에테르 단량체들의 합성과 광중합 특성)

  • Kim, Ki-Sang;Shim, Sang-Yeon
    • Journal of the Korean Applied Science and Technology
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    • v.34 no.2
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    • pp.203-209
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    • 2017
  • The propenyl ether-type monomers which are applicable for cationic photo-polymerization were synthesized by the condensation reaction of mono and di-functional alcohol with allyl bromide. To examine photo-curable reactivity, these monomers were combined with cationic photoinitiator to prepare coating composition. As a result, the initial rate of polymerization of POMB in mono propenyl ether type was 10.2, which was relatively lower than BPOB in di-propenyl ethers type. However, POMB containing 1.5mol% photoinitiator almost quantitatively reacted within 90 seconds. In addition, Sulfonium salt type photo-initiators containing long-alkyl group showed good solubility with monomers and apperaed to have comparatively higher rate of polymerization and conversion ratio when applying DPSA and DPST which have high acidity on all monomers.

Characterization of Porous Asphalt Concrete Coated with MMA Resin (아크릴 수지로 코팅한 배수성 아스팔트 콘크리트의 특성 평가)

  • Choi, Tae-Jun;Lee, Hyun-Jong;Kim, Tae-Woo;Song, Jae-Hyok
    • International Journal of Highway Engineering
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    • v.11 no.2
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    • pp.111-120
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    • 2009
  • This paper presents a fundamental findings of the functional and structural performance of the porous asphalt concrete coated with MMA resin. To evaluate the structural performance, cantabro, wheel tracking, moisture sensitivity and indirect tensile fatigue tests are performed. The tests results show that the cantabro loss is reduced three times and fatigue resistance is significantly increased after the specimens are coated with MMA resin. However there are little changes in the rutting and moisture damage resistances before and after the coating. Air voids, permeability and BPT(British Pendulum Test) tests are conducted to study the functional performance. It is observed form the tests that the air voids and permeability are slightly decreased after the coating. However, the changes in the air voids and permeability are negligible. The skid resistance of the coated specimens is lower than reference specimens. However, the skid resistance is maintained beyond the level of the reference specimens when silica sands are chipped on top of the coated surface.

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Characterization of Double-layer Optical Sensing Membranes for Dual Sensing of Dissolved Oxygen Concentrations and pH and Their Application to the On-line Monitoring of Fermentation Processes (용존산소농도와 pH의 동시 검출용 이중층 광학 센서막의 특성 및 발효공정의 온라인 모니터링에의 응용)

  • Kim, Chun-Kwang;Lee, Jong-Il
    • Applied Chemistry for Engineering
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    • v.20 no.4
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    • pp.370-374
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    • 2009
  • A double-layer optical sensing membrane has been fabricated to measure the concentration of dissolved oxygen (DO) and pH value simultaneously. (tris(4,7-diphenyl-1,10-phenanthroline (Rudpp) ruthenium(II)) as a DO sensitive dye has been mixed in the methyl trimethoxy silane (MTMS) sol-gel solution and coated onto one well in a 24-well microtiter plate. On the DO-sensing layer the GA (3-glycidoxypropyl trimethoxy silane (GPTMS), 3-aminopropyl trimethoxy silane (APTMS)) sol-gel solution mixed with 8-hydroxypyrene-1,3,6-trisulfonic acid trisodium salt (HPTS) has been coated and used to measure pH values. The double-layer sensing membrane was affected by ionic strength and temperature. The double-layer sensing membrane for DO and pH has been applied to online monitor in microorganism cultivation processes and showed a good performance.

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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Characterization of Shelf Life Extension Packaging Material for Food and Fresh Cut Agricultural Product: A Review (식품, 신선편이 농산물용 저장기간 연장 포장 소재 특성 및 평가)

  • Rhee, Jin-Kyu;Yu, Ji Ye;Kim, Mi-Kyung;You, Young-Sun
    • KOREAN JOURNAL OF PACKAGING SCIENCE & TECHNOLOGY
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    • v.22 no.3
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    • pp.119-125
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    • 2016
  • Fresh-cut agricultural products provide convenience to consumers. However, quality changes or microbial growth can easily occur due to physical changes such as cutting and peeling etc. during processing. Therefore, efforts have been made to develop a functional packaging for extension of shelf life of fresh-cut agricultural products, food etc., and researches on prolongation of storage period have been actively developed. The shelf life is extended by antimicrobial, far infrared rays, air permeability, anti-fogging, weak current, ethylene gas adsorption or decomposition, gas composition changes such as MA (Modified Atmosphere) or CA (Controlled Atmosphere). The method of extending the shelf life by various complex factors. This paper based on the published literature to extend the shelf life of fresh-cut agricultural products, food etc., The paper has summarized the storage period extension packing method, packaging material for shelf life extension and comprehensive evaluation method.

Cloning and Characterization of cDNA Encoding Potentially Functional Mouse Glandular Kallikrein

  • Kim, Hwa-Seon;Kim, Won-Sin
    • BMB Reports
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    • v.30 no.5
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    • pp.356-361
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    • 1997
  • We cloned a cDNA (pPRC-1) which was comprised of 841 nucleotides from the cDNA library of a male ICR mouse submandibular gland ($SMG^+$). The nucleotide sequences of pPRC-1 were identical to those of exons 2 and 3 of the mGK-21 gene, a potentially functional glandular kallikrein identified in a Balb/c mouse, except for one nucleotide residue. Although this substitution changes Ile (ATT) in pPRC-1 to Val (GTT) in mGK-21, this difference has been explained by strain polymorphism. From the amino acid sequences predicted from its cDNA, we speculated that mGK-21 gene products/pGK21 consist of 261 amino acids including the $NH_2$-terminal signal peptide (residues 1~17), the short propeptide (residues 17~24), and the active peptide (residues 25~261). Although we did not demonstrate the enzyme activity of pGK21, it was assumed that pGK 21 was involved in the maturation of certain bioactive polypeptide(s) in mouse SMG for the following reasons : (a) mGK-21 gene was apparently expressed in a male ICR mouse SMG: (b) the proposed active site $His^{65}$, $Asp^{120}$, and $Ser^{213}$ residues were completely conserved in pGK21 just like other glandular kallikreins; (c) the cloned cDNA was translated to a predicted 27 kDa polypeptide chain in vitro: (d) the 27 kDa polypeptide chain produced by CHO cells was produced to a putative active form by trypsin.

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