• Title/Summary/Keyword: structure elucidation

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Analysis of Structure and Expression of Grapevine 2-oxoglutarate Oxygenase Genes in Response to Low Temperature

  • Kim, Seon Ae;Ahn, Soon Young;Yun, Hae Keun
    • Horticultural Science & Technology
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    • v.34 no.1
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    • pp.46-54
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    • 2016
  • 2-Oxoglutarate (2OG) acts as a signaling molecule and plays a critical role in secondary metabolism in a variety of organisms, including plants. Six 2-oxoglutarate (2OG) and Fe(II) oxygenase (2OGO) genes, VlCE2OGO1 [Vitis labruscana 2-oxoglutarate (2OG) and Fe(II) oxygenase 1], VlCE2OGO2, VlCE2OGO3, VlCE2OGO4, VlCE2OGO5, and VlCE2OGO6, which show different expression patterns upon transcriptome analysis of 'Campbell Early' grapevine exposed to low temperature for 4 weeks, were analyzed for their structure and expression. Comparison of the deduced amino acid sequences of the 2OGO genes from the V. labruscana transcripts revealed sequence similarities of 38.6% (VlCE2OGO1 and VlCE2OGO2) to 19.2% (VlCE2OGO2 and VlCE2OGO3). The lengths of these genes ranged from 1053 to 2298 bp, and they encoded 316 to 380 amino acids. The prediction of the secondary structure of the encoded proteins by Self-Optimized Prediction Method with Alignment (SOPMA) indicated that all the genes contained alpha helix (23.95 to 41.71%), extended strand (16 to 22.34%), beta turn (6.65 to 9.22%), and random coil (32.97 to 51.58%) in the analysis. Specific primers from unique regions in each gene obtained by alignment of nucleotide sequences were used in real time PCR for analysis of gene expression. All tested genes showed differential expression in grapevines exposed to low temperature. Of the six transcripts, VlCE2OGO1, VlCE2OGO2, and VlCE2OGO3 were up-regulated and VlCE2OGO4, VlCE2OGO5, and VlCE2OGO6 were down-regulated in response to cold treatments at all tested time points. The 2OG genes can be used for elucidation of mechanisms of tolerance to cold and as valuable molecular genetic resources for selection in breeding programs for cold-hardy grapevines.

Gene Expression Analysis of Lung Injury in Rats Induced by Exposure to MMA-SS Welding Fume for 30 Days

  • Oh, Jung-Hwa;Park, Han-Jin;Heo, Sun-Hee;Yang, Mi-Jin;Yang, Young-Su;Song, Chang-Woo;Yoon, Seok-Joo
    • Molecular & Cellular Toxicology
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    • v.3 no.4
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    • pp.306-313
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    • 2007
  • The welding fume has been implicated as a causal agent in respiratory disease such as pneumoconiosis. The molecular mechanism by which welding fume induces toxicity in the lung is still unknown, but studies have focused on histological structure and indirect approach measuring the pulmonary damage markers. In the present study, gene expression profiles were analyzed in the lung of rats exposed by manual metal-arc stainless-steel (MMA-SS) welding fume for 30 days using Affymetrix GeneChip$^{(R)}$. Totally, 379 genes were identified as being either up- or down-regulated over 2-fold changes (P<0.01) in the lung of low- or high-dose group and were analyzed by using hierarchical clustering. We focused on genes involved in immune/inflammation responses were differentially regulated during lung injury induced by welding fume exposure. The information of these deregulated genes may contribute in elucidation of the inflammation mechanism during lung injury such as lung fibrosis.

Prediction of Genes Lacking in an Ammonia Oxidizing Archaeon for Independent Growth (암모니아 산화 고세균의 독립성장에 필요한 결손 유전자 예측)

  • Han, Sang-Soo;Lee, Jin-Young;Rhee, Sung-Keun;Kim, Geun-Joong
    • KSBB Journal
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    • v.26 no.3
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    • pp.237-242
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    • 2011
  • As a number of archaea are ubiquitously found in non-extreme habitats, elucidation of their functional roles becomes currently an emerging issue. However, most of them are unable to grow in pure culture and so it remains to be established. In order to find genes lacking in the genome of an ammonia-oxidizing archaeon (AOA), we here report on the comparative analyses of an AOA genome with those of experimentally or theoretically established minimal genomes for independent growth. We assessed the genes lacking in AOA using logic of clusters of orthologous groups (COG), remote homology, consensus sequence weight matrix, function-based motif or domain, and then further excluded genes encoding hypothetical orarchaea-specific proteins. The results of these combination analyses revealed 19 candidate genes lacking in the genome of an AOA. Thus, our results provide a possibility of inducing independent growth of AOA when supplemented with product (s) of the lacking gene (s), and also give a chance for finding new proteins with novel sequence or structure space even if the predicted lacking-genes will be found using another algorithms or biochemical studies.

Metabolites of Marine Algae Collected from Karachi-coasts of Arabian Sea

  • Ali, Muhammad Shaiq;Jahangir, Muhammad;Saleem, Muhammad;Pervez, Muhammad Kashif;Hameed, Shaista;Ahmad, Viqar Uddin
    • Natural Product Sciences
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    • v.6 no.2
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    • pp.61-65
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    • 2000
  • The ethanolic extracts of marine green, brown and red algae collected from Karachi coasts of Arabian Sea afforded a new enol-derivative of N-acylsphingosine named as coelarthenol (1) from Coelarthrum muelleri, two new glucose-derivatives named: botryenal (2) and botryenol (3) from Botryocladia leptopoda, ${\alpha}-tocopherol$ quinone (4) from Codium iyengarii, ${\beta}-sitosterol$ and hexadecanoic acid from Stokeyia indica. The known constituents (4, ${\beta}-sitosterol$ & hexadecanoic acid) have not been reported so far from their corresponding sources and the structures were determined through spectroscopic methods, whereas, the structures of new constituents (1-3) were elucidated with the aid of selective HMBC experiments. The phytotoxicity of 4 was also monitored.

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Citrinin Hydrate Inhibit Serotonin N-Acetyltransferase Catalyzing the Conversion of Serotonin to N-Acetylserotonin

  • Lee, In-Kyoung;Yun, Bong-Sik;Kim, Kyong-Tai;Choi, Bo-Hwa;Park, Tae-Ju;Kim, Young-Ho;Yoo, Ick-Dong
    • Journal of Microbiology and Biotechnology
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    • v.11 no.6
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    • pp.1099-1101
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    • 2001
  • In an attempt to search for serotonin N-acetyltransferase (arylalkylamine N-acetyltransferasem, AA-NAT) inhibitors from microbial metabolites, we fecund the culture broth of Penicillium sp. 80722 which showed a strong inhibitory activity against AA-NNT. The active principle has been identified as citrinin hydrate through bioassay-guided fractionation of cultural broth, and structure elucidation derived by spectroscopic analyses. Citrinin hydrate inhibits AA-NAT with an $IC_50$ value of $173{\mu}M$ in a dose-dependent manner. Although citrinin hydrate was previously isolated as human rhinovirus 3C-protease inhibitor, this was recognized as the first AA-NAT inhibitor isolated from natural sources.

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Structural Characteristics of Cell Walls of Forage Grasses - Their Nutritional Evaluation for Ruminants - - Review -

  • Iiyama, Kenji;Tuyet Lam, Thi Bach
    • Asian-Australasian Journal of Animal Sciences
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    • v.14 no.6
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    • pp.862-879
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    • 2001
  • The walls of all higher plants are organized as a cellulosic, fibrillar phase embedded in a matrix phase composed of non-cellulosic polysaccharides, some proteins and, in most secondary walls, lignin. At the effective utilization of plant biomass, qualitative and quantitative analyses of plant cell walls are essential. Structural features of individual components are being clarified using newly developed equipments and techniques. However, "empirical" procedures to elucidate plant cell walls, which are not due to scientific definition of components, are still applied in some fields. These procedures may give misunderstanding for the effective utilization of plant biomass. In addition, interesting the investigation of wall organization is moving towards not only qualitatively characterisation, but also quantitation of the associations between wall components. These involve polysaccharide-polysaccharide and polysaccharide-lignin cross-links. Investigation of the associations is being done in order to understand the chemical structure, organization and biosynthesis of the cell wall and physiology of the plants. Procedures for qualitative and quantitative analyses based on the definition of cell wall components are reviewed focussing in nutritional elucidation of forage grasses by ruminant microorganisms.

Engineering Hybrid Proteins by Modular Recombination and Evolutionary Optimization (모듈성 단백질의 재설계 및 개량)

  • Lee, Seung-Goo;Rha, Eu-Gene;Ha, Jae-Seok;Lee, Jeong-Min;Kim, Sun-Hwa
    • Microbiology and Biotechnology Letters
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    • v.36 no.2
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    • pp.149-157
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    • 2008
  • Many proteins consist of distinctive domains that can act independently or cooperatively to achieve a unique function. As these domains evolve from a naturally existing repertoire of functional domains, this implies that domain organization is an intrinsic element involved in building the complex structure and function of proteins. Thus, identifying functional domains would appear to be critical to the elucidation of questions related to protein evolution, folding, and the engineering of hybrid proteins for tai- lored applications. However, the simple application of "Lego-like assembly" to the engineering of hybrid proteins is an oversimplification, as many hybrid constructs lack structural stability, usually due to unfavorable domain contacts. Thus, directed evolution, along with computational studies, may help to engineer hybrid proteins with improved physico-chemical properties. Accordingly, this paper introduces several approaches to functional hybrid protein engineering that potentially can be used to create modulators of gene transcription and cell signaling, and novel biosensors to analyze biological functions in vivo.

Local Climate Mediates Spatial and Temporal Variation in Carabid Beetle Communities on Hyangnobong, Korea

  • Park, Yong Hwan;Jang, Tae Woong;Jeong, Jong Cheol;Chae, Hee Mun;Kim, Jong Kuk
    • Journal of Forest and Environmental Science
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    • v.33 no.3
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    • pp.161-171
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    • 2017
  • Global environmental changes have the capacity to make dramatic alterations to floral and faunal composition, and elucidation of the mechanism is important for predicting its outcomes. Studies on global climate change have traditionally focused on statistical summaries within relatively wide scales of spatial and temporal changes, and less attention has been paid to variability in microclimates across spatial and temporal scales. Microclimate is a suite of climatic conditions measured in local areas near the earth's surface. Environmental variables in microclimatic scale can be critical for the ecology of organisms inhabiting there. Here we examine the effect of spatial and temporal changes in microclimates on those of carabid beetle communities in Hyangnobong, Korea. We found that climatic variables and the patterns of annual changes in carabid beetle communities differed among sites even within the single mountain system. Our results indicate the importance of temporal survey of communities at local scales, which is expected to reveal an additional fraction of variation in communities and underlying processes that has been overlooked in studies of global community patterns and changes.

Characterization of the active site and coenzyme binding pocket of the monomeric UDP- galactose 4'- epimerase of Aeromonas hydrophila

  • Agarwal, Shivani;Mishra, Neeraj;Agarwal, Shivangi;Dixit, Aparna
    • BMB Reports
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    • v.43 no.6
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    • pp.419-426
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    • 2010
  • Aeromonas hydrophila is a bacterial pathogen that infects a large number of eukaryotes, including humans. The UDP-galactose 4'-epimerase (GalE) catalyzes interconversion of UDP-galactose to UDP-glucose and plays a key role in lipopolysaccharide biosynthesis. This makes it an important virulence determinant, and therefore a potential drug target. Our earlier studies revealed that unlike other GalEs, GalE of A. hydrophila exists as a monomer. This uniqueness necessitated elucidation of its structure and active site. Chemical modification of the 6xHis-rGalE demonstrated the role of histidine residue in catalysis and that it did not constitute the substrate binding pocket. Loss of the 6xHis-rGalE activity and coenzyme fluorescence with thiol modifying reagents established the role of two distinct vicinal thiols in catalysis. Chemical modification studies revealed arginine to be essential for catalysis. Site-directed mutagenesis indicated Tyr149 and Lys153 to be involved in catalysis. Use of glycerol as a cosolvent enhanced the GalE thermostability significantly.

Isolation and structure elucidation of antifungal compounds from the antarctic lichens, Stereocaulon alpinum and Sphaerophorus globosus

  • Kim, Young-Shin;Lim, Chi-Hwan
    • Korean Journal of Agricultural Science
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    • v.47 no.1
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    • pp.183-191
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    • 2020
  • Lichens are composite organisms consisting of a symbiotic association of a fungus with a photosynthetic partner (the photobiont or phycobiont), usually either a green alga or cyanobacterium. According to more recent studies, the biological activities of lichens and lichen substances include an antibiotic activity, antitumor and antimutagenic activity against human immunodeficiency virus (HIV), allergenic activity, plant growth inhibitory activity, and enzyme inhibitory activity. This study screened lichen extracts with a potent in vitro antifungal activity against plant diseases caused by phytopathogenic fungi. The compounds were isolated from Stereocaulon alpinum and Sphaerophorus globosus, and their chemical structures were identified as methyl hematommate, methyl β-orsellinate, 5-hydroxyferulic acid, sphaerophorin, and 2-heptyl-4,6-dimethoxybenzoic acid by electron ionization mass spectrometry (EI-MS) and nuclear magnetic resonance (NMR) spectral analyses. In vitro disease control against Alternaria mali, Cochliobolus miyabeanus, Colletotrium gloeosporioides, and Verticillum dahliae was evaluated. And among the five compounds, only methyl hematommate was effective against A. mali, C. miyabeanus, and C. gloeosporioides. The compounds were isolated from these lichens, which have a similar biosynthetic pathway, respectively. This is the first report of these compounds being isolated from these lichens.