• Title/Summary/Keyword: Functional characterization

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Structural and Functional Characterization of CRAMP-18 Derived from a Cathelicidin-Related Antimicrobial Peptide CRAMP

  • Park, Kyong-Soo;Shin, Song-Yub;Hahm, Kyung-Soo;Kim, Yang-Mee
    • Bulletin of the Korean Chemical Society
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    • v.24 no.10
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    • pp.1478-1484
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    • 2003
  • CRAMP was identified from a cDNA clone derived from a mouse femoral marrow cells as a member of cathelicidin-derived antimicrobial peptide. Tertiary structure of CRAMP in TFE/$H_2O$ (1 : 1, v/v) solution has been determined by NMR spectroscopy previously and consists of two amphipathic $\alpha-helices$ from Leu4 to Lys10 and from Gly16 to Leu33. These two helices are connected by a flexible region from Gly11 to Gly16. Analysis of series of fragments composed of various portion of CRAMP revealed that an 18-residue fragment with the sequence from Gly16 to Leu33 (CRAMP-18) was found to retain antibacterial activity without cytotoxicity. The effects of two Phe residues at positions 14 and 15 of CRAMP-18 on structure, antibacterial activity, and interaction with lipid membranes were investigated by $Phe^{14,15}$ ${\rightarrow}$ Ala substitution (CRAMP-18-A) in the present study. Substitution of Phe with Ala in CRAMP-18 caused a significant reduction on antibacterial and membrane-disrupting activities. Tertiary structures of CRAMP-18 in 50% TFE/$H_2O$ (1 : 1, v : v) solution shows amphipathic ${\alpha}$-helix, from $Glu^2{\;}to{\;}Leu^{18}$, while CRAMP-18-A has relatively short amphipathic ${\alpha}$-helix from $Leu^4{\;}to{\;}Ala^{15}$. These results suggest that the hydrophobic property of $Phe^{14}{\;}and{\;}Phe^15$ in CRAMP-18 is essential for its antibacterial activity, ${\alpha}$-helical structure, and interactions with phospholipid membranes.

Functional Characterization and Regional Expression of Dopamine Transporter (도파민 수송체의 기능적 특성 및 발현에 관한 연구)

  • 이상훈;이송득;성기욱;이동섭;이용성;고재경
    • YAKHAK HOEJI
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    • v.39 no.2
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    • pp.161-168
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    • 1995
  • Brain dopamine systems play a central role in the control of movement, hormone release, and many complex behavior. The action of dopamine at its synapse is terminated predominately by high affinity reuptake into presynaptic terminals by dopamine transporter (DAT). The dopamine transporter(DAT) is membrane protein localized to dopamine-containing nerve terminals and closely related with cocaine abuse, Parkinsonism, and schizophrenia. In present study, the recombinant plasmid pRc/CMV-DAT, constructed by subcloning of a cDNA encoding a bovine DAT into eukaryotic expression vector pRc/CMV, was stably transfected into CV-1 cells(monkey kidney cell line). The DAT activities in the cell lines selected by Geneticin$^{R}$ were determined by measuring the uptake of $[^3H]$-dopamine. The transfected cell lines showed 30-50 fold higher activities than untransfected CV-1 cell line, and this result implies that DAT is well expressed and localized in transfected cells. The transfected cells accumulated $[^3H]$-dopamine in a dose-dependent manner with a $K_{m}$ of 991.6nM. Even though high doses of norepinephrine, epinephrine, serotonin, and choline neurotransmitters inhibited the uptake of $[^3H]$-dopamine, DAT in transfected cell line was proven to be much more specific to dopamine. The psychotropic drugs such as GBR12909, CFT, normifensine, clomipramine, desipramine, and imipramine inhibited significantly the dopamine uptake in tissue culture cells stably transfected with DAT cDNA. Radioactive in situ hybridization was done to map the cellular localization of DAT mRNA-containing cells in the adult rat central nervous system. The strong hybridization signals were detected only in the substantia nigra pars compacta and ventral tegmental area. The restricted anatomical localization of DAT mRNA-containing cells confirms the DAT as a presynaptic marker of dopamine-containing cells in the rat brain.

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Sonocatalytic Degradation of Rhodamine B in the Presence of TiO2 Nanoparticles by Loading WO3

  • Meng, Ze-Da;Sarkar, Sourav;Zhu, Lei;Ullah, Kefayat;Ye, Shu;Oh, Won-Chun
    • Korean Journal of Materials Research
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    • v.24 no.1
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    • pp.6-12
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    • 2014
  • In the present work, $WO_3$ and $WO_3-TiO_2$ were prepared by the chemical deposition method. Structural variations, surface state and elemental compositions were investigated for preparation of $WO_3-TiO_2$ sonocatalyst. X-ray diffraction (XRD), scanning electron microscopy (SEM), energy dispersive X-ray (EDX) and transmission electron microscopy (TEM) were employed for characterization of these new photocatalysts. A rhodamine B (Rh.B) solution under ultrasonic irradiation was used to determine the catalytic activity. Excellent catalytic degradation of an Rh.B solution was observed using the $WO_3-TiO_2$ composites under ultrasonic irradiation. Sonocatalytic degradation is a novel technology of treating wastewater. During the ultrasonic treatment of aqueous solutions sonoluminescence, cavitaties and "hot spot" occurred, leading to the dissociation of water molecules. In case of a $WO_3$ coupled system, a semiconductor coupled with two components has a beneficial role in improving charge separation and enhancing $TiO_2$ response to ultrasonic radiations. In case of the addition of $WO_3$ as new matter, the excited electrons from the $WO_3$ particles are quickly transferred to $TiO_2$ particle, as the conduction band of $WO_3$ is 0.74 eV which is -0.5 eV more than that of $TiO_2$. This transfer of charge should enhance the oxidation of the adsorbed organic substrate. The result shows that the photocatalytic performance of $TiO_2$ nanoparticles was improved by loading $WO_3$.

Improvement of Wettability and Removal of Skin Layer on Ar-Plasma-Treated Polypropylene Blend Surface (폴리프로필렌 복합소재의 아르곤 플라즈마 처리로 표면층 제거와 젖음성 향상)

  • Weon, Jong-Il;Lee, Sun-Yong
    • Polymer(Korea)
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    • v.36 no.4
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    • pp.461-469
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    • 2012
  • The surface modification and characterization of Ar-plasma treated polypropylene (PP) blend are investigated using x-ray photoelectron spectroscopy (XPS), Fourier transform infrared spectroscopy (FTIR), scanning electron microscopy (SEM) and contact angle measurement. An increase in Ar-plasma treatment time leads to an increase in wettability, oxygen containing polar functional groups, the amount of talc, and surface roughness on the PP blend surface. A careful observation using SEM indicates that there exists a skin layer consisting of only PP component. The difference in viscosity between PP and rubber particles facilities the formation of skin layer. However, it is found that an increase in Ar-plasma treatment time helps to decrease the thickness of skin layer. Additional methodologies for the elimination of skin layer during injection molding are also discussed. The surface modification and morphological alteration induced by Ar-plasma treatment provides a hydrophilic state, followed by the improvement in wettability, on the PP blend surface.

Quantitative Evaluation of Scratch Behavior for Polymeric Materials (고분자 소재의 스크래치 거동의 정량적 평가)

  • Baek, Ki-Wan;Lee, Sung-Goo;Lee, Jae-Heung;Choi, Kil-Yeong;Weon, Jong-Il
    • Polymer(Korea)
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    • v.33 no.4
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    • pp.273-283
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    • 2009
  • Recent research issues on the scratch behaviors of polymeric materials has been investigated. In this study, the scratch characterization of polymeric materials with respect of experimental parameters, such as nature of the material, temperature, applied load, test speed, surface treatment, scratch number of times, polymer structure/functional groups, degree of cross-linking, and crystallinity, are reviewed. In addition, the testing standards and methodologies which could quantify the scratch behaviors are introduced and the current international standards are compared and summarized. The latest technical approaches for evaluating the scratch behaviors and improving the scratch resistance of polymers are also discussed.

Overexpression of ginseng patatin-related phospholipase pPLAIIIβ alters the polarity of cell growth and decreases lignin content in Arabidopsis

  • Jang, Jin Hoon;Lee, Ok Ran
    • Journal of Ginseng Research
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    • v.44 no.2
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    • pp.321-331
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    • 2020
  • Background: The patatin-related phospholipase AIII family (pPLAIIIs) genes alter cell elongation and cell wall composition in Arabidopsis and rice plant, suggesting diverse commercial purposes of the economically important medicinal ginseng plant. Herein, we show the functional characterization of a ginseng pPLAIII gene for the first time and discuss its potential applications. Methods: pPLAIIIs were identified from ginseng expressed sequence tag clones and further confirmed by search against ginseng database and polymerase chain reaction. A clone showing the highest homology with pPLAIIIβ was shown to be overexpressed in Arabidopsis using Agrobacterium. Quantitative polymerase chain reaction was performed to analyze ginseng pPLAIIIβ expression. Phenotypes were observed using a low-vacuum scanning electron microscope. Lignin was stained using phloroglucinol and quantified using acetyl bromide. Results: The PgpPLAIIIβ transcripts were observed in all organs of 2-year-old ginseng. Overexpression of ginseng pPLAIIIβ (PgpPLAIIIβ-OE) in Arabidopsis resulted in small and stunted plants. It shortened the trichomes and decreased trichome number, indicating defects in cell polarity. Furthermore, OE lines exhibited enlarged seeds with less number per silique. The YUCCA9 gene was downregulated in the OE lines, which is reported to be associated with lignification. Accordingly, lignin was stained less in the OE lines, and the expression of two transcription factors related to lignin biosynthesis was also decreased significantly. Conclusion: Overexpression of pPLAIIIβ retarded cell elongation in all the tested organs except seeds, which were longer and thicker than those of the controls. Shorter root length is related to auxinresponsive genes, and its stunted phenotype showed decreased lignin content.

Generation and DNA Characterization of High-lysine Mutants by Biochemical Selection from Callus Culture of 'Hwayeongbyeo'

  • Yi Gi-Hwan;Choi Jun-Ho;Kim Kyung-Min;Jeong Eung-Gi;Park Hyang-Mi;Kim Doh-Hoon;Ku Yeon Chung;Eun Moo-Young;Kim Ho-Yeong;Nam Min-Hee
    • Plant Resources
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    • v.8 no.1
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    • pp.60-66
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    • 2005
  • Lysine is the first essential amino acid for optimal nutrient quality in rice grain. For the narrow genetic diversities of lysine contents in rice, somaclonal variation was the source of mutation in our breeding program. Biochemical selection was conducted using 1 mM S-(2-aminoethyl) cysteine followed by two passages of 5 mM lysine plus threonine in the callus subculture medium. The lysine contents in endosperm of all progenies recovered from the biochemical selection were higher than those of their donor cultivar 'Hwayeongbyeo'. These elevated lysine levels of mutants were successfully transmitted to $M_4$ generation. The lysine contents in endosperm varied 3.85 to $4.80\%$ compare to their donor cultivar 'Hwayeongbyeo' was $3.85\%$. Three of high-lysine germplasms, Lys-l, Lys-2 and Lys-7 were selected by biochemical selection and rapid screening methods. DNA analysis showed that a new insertion of Tos 17 which mapped to rice chromosome 11 on the high-lysine mutant, Lys-2.

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Isolation and Characterization of Nucleoside Diphosphate Kinase 1 of Codonopsis lanceolata (더덕에서 Nucleoside Diphosphate Kinase 1 분리 및 분석)

  • 김종학;양덕춘
    • Korean Journal of Plant Resources
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    • v.16 no.3
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    • pp.257-263
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    • 2003
  • The NDK1 is an ubiquitous enzyme that transfer phosphate groups from triphosphate nucleoside diphosphates(NDPs) in eukaryotes and prokaryotes. We isolated and characterized a cDNA encoding a nucleoside diphosphate kinase 1(CNDK 1) in Codonopsis lanceolata. The CNDK 1 is 444bp long and open reading frame of 447bp with a deduced amino acid of 148 residue. The CNDK 1 has an ATP binding site in 12­16 residue and phosphohistidine intermediate in 115 residue of amino acid sequence. Although several NDK 1 genes have been cloned in plants, but little is known about the functional significance of this enzyme during plant growth and development. The CNDK 1 shows the identities to Arabidopsis thaliana (71%), Oryza sativa(75%), Glycine max (79%), Brassica rapa (77%), Mesembryanthemum crystallinum (85 %), Spinacia oleracea (83%), Pisum sativum (82%). The CNDK 1 of C. laceolata have a closer relationship of Glycine max and Pisum sativum at the phylogenic analysis.

Synthesis and Characterization of Mono- and Diacylglycerol Enriched Functional Oil by Enzymatic Glycerolysis of Corn Oil (옥수수유로부터의 효소적 glycerolysis에 의한 monoacylglycerol과 diacylglycerol 함유 기능성 유지 합성 및 특성연구)

  • Park, Rae-Kyun;Lee, Ki-Teak
    • Korean Journal of Food Science and Technology
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    • v.36 no.2
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    • pp.211-216
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    • 2004
  • Mono-and diacylglycerol-enriched oil was produced from corn oil through enzymatic glycerolysis using 1,3-specific immobilized lipase in solvent-free system and stirred-tank batch reactor. HPLC analysis revealed enriched oil was respectively composed of: 45.05, 16.27, 23.05, and 14.98% triacylglycerol, 1,3-diacylglycerol, 1,2-diacylglycerol, and monoacylglycerol; 13.21, 0.15, 2.02, 34.36, 49.12, and 1.14 mol% palmitic, palmitoleic, stearic, oleic, linoleic, and linolenic acids; and 0.014, 0.029, 0.010 and 0.053% ${\alpha},\;{\gamma},\;{\delta}-$, and total tocopherols. Physiochemical and melting properties of enriched oil were evaluated. Oxidative stability study revealed enriched oil showed higher peroxide and p-anisidine values than corn oil. Rosemary extracts (100 to 300 ppm) reduce oxidation.

A Cytosolic Thioredoxin Acts as a Molecular Chaperone for Peroxisome Matrix Proteins as Well as Antioxidant in Peroxisome

  • Du, Hui;Kim, Sunghan;Hur, Yoon-Sun;Lee, Myung-Sok;Lee, Suk-Ha;Cheon, Choong-Ill
    • Molecules and Cells
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    • v.38 no.2
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    • pp.187-194
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    • 2015
  • Thioredoxin (TRX) is a disulfide reductase present ubiquitously in all taxa and plays an important role as a regulator of cellular redox state. Recently, a redox-independent, chaperone function has also been reported for some thioredoxins. We previously identified nodulin-35, the subunit of soybean uricase, as an interacting target of a cytosolic soybean thioredoxin, GmTRX. Here we report the further characterization of the interaction, which turns out to be independent of the disulfide reductase function and results in the co-localization of GmTRX and nodulin-35 in peroxisomes, suggesting a possible function of GmTRX in peroxisomes. In addition, the chaperone function of GmTRX was demonstrated in in vitro molecular chaperone activity assays including the thermal denaturation assay and malate dehydrogenase aggregation assay. Our results demonstrate that the target of GmTRX is not only confined to the nodulin-35, but many other peroxisomal proteins, including catalase (AtCAT), transthyretin-like protein 1 (AtTTL1), and acyl-coenzyme A oxidase 4 (AtACX4), also interact with the GmTRX. Together with an increased uricase activity of nodulin-35 and reduced ROS accumulation observed in the presence of GmTRX in our results, especially under heat shock and oxidative stress conditions, it appears that GmTRX represents a novel thioredoxin that is co-localized to the peroxisomes, possibly providing functional integrity to peroxisomal proteins.