• Title/Summary/Keyword: Second cleavage

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Mini-review: oomycete RXLR genes as effector-triggered immunity

  • Arif, Saima;Jang, Hyun A;Kim, Mi-Reu;Oh, Sang-Keun
    • Korean Journal of Agricultural Science
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    • 제45권4호
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    • pp.561-573
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    • 2018
  • Oomycetes are known to secrete a vast arsenal of effectors that modulate the host defense system as well as facilitate establishing a parasitic infection in plants. In recent years, tremendous progress has been made in the field of effectromics based on studies of oomycetes, especially the cytoplasmic family of RXLR effectors. Yet, the biology of the RXLR effector family is still poorly understood. There has been a consensus regarding the structure of the RXLR motif in the mycologist community. However, the function of the RXLR motif is still unclear. First, different models have suggested that the role of the RXLR motif is either in translocation to a target destination inside a host cell or in the cleavage of itself followed by secretion. Second, recent studies have suggested different functional models for the RXLR motif. According to a widely accepted model, the RXLR motif is directly involved in the translocation of effectors to target sites. In contrast, a new study has proposed that the RXLR motif is involved in secretion rather than translocation. Thus, this review is an attempt to summarize the recent advances made in the functional analysis of the N-terminal domain of RXLR effectors.

Flavonoids Biotransformation by Human Gut Bacterium Dorea sp. MRG-IFC3 Cell-Free Extract

  • Huynh Thi Ngoc Mi;Heji Kim;Jong Suk Lee;Bekir Engin Eser;Jaehong Han
    • Journal of Microbiology and Biotechnology
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    • 제34권6호
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    • pp.1270-1275
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    • 2024
  • Human gut bacterium Dorea sp. MRG-IFC3 is unique in that it is capable of metabolizing puerarin, an isoflavone C-glycoside, whereas it shows broad substrate glycosidase activity for the various flavonoid O-glycosides. To address the question on the substrate specificity, as well as biochemical characteristics, cell-free biotransformation of flavonoid glycosides was performed under various conditions. The results showed that there are two different enzyme systems responsible for the metabolism of flavonoid C-glycosides and O-glycosides in the MRG-IFC3 strain. The system responsible for the conversion of puerarin was inducible and comprised of two enzymes. One enzyme oxidizes puerarin to 3"-oxo-puerarin and the other enzyme converts 3"-oxo-puearin to daidzein. The second enzyme was only active toward 3"-oxo-puerarin. The activity of puerarin conversion to daidzein was enhanced in the presence of Mn2+ and NAD+. It was concluded that the puerarin C-deglycosylation by Dorea sp. MRG-IFC3 possibly adopts the same biochemical mechanism as the strain PUE, a species of Dorea longicatena.

Early Life History of Black Bullhead, Pseudobagrus koreanus(Pisces, Bagridae), from Kum River, Korea (금강에 서식하는 눈동자개 Pseudobagrus koreanus(Pisces, Bagridae)의 초기 생활사)

  • Kang, Eon-Jong
    • Korean Journal of Ichthyology
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    • 제10권2호
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    • pp.184-190
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    • 1998
  • The early life history of black bullhead, Pseudobagrus koreanus, endemic to Korea was investigated to get biological information needed in artificial production of seedlings and in recovering natural resources. The fertilized eggs showed some characteristics in having heavy sticky material and minute folds which is formed radical pattern on the egg membrane. The shape of egg was spherical and $2.59{\pm}0.08$(2.45~2.70, n=10)mm in diameter. The yolk had not oil globule. The first cleavage was observed 2 hrs after insemination at $21{\sim}23^{\circ}C$, and the progressive cleavage were done about 30 min. interval. The characteristic changing of the yolk surface started at morula stage and continued to the end of gastrula. Hatching was started 72 hrs and completed 90 hrs after fertilization. The size of the larvae were 5.41~5.81mm in total length and 2.76~2.94mm in preanal length, and the number of so mites was 15-16+33~34(48~50). The barbels and swimbladder were completed and all the fins except second dorsal were appeared 1 week after hatching. The larvae attained 9.67~10.52mm in total length and 5.20~5.65mm in preanal length. All the fin sets and color pattern were completed 2 weeks after hatching and body mucus was secreted at that stage. The juvenile attained 14.59~16.02mm in standard length, 3.31~4.16mm in head length and 8.07~9.31mm in prenal length.

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Effects of Fructose in a Chemically Defined Maturation Medium on Oocyte Maturation and Parthenogenetic Embryo Development in Pigs (돼지 난자의 체외성숙에서 합성배양액에 첨가된 과당이 난자의 성숙 및 단위발생 배아의 체외발육에 미치는 영향)

  • Shin, Hyeji;Kim, Minji;Lee, Joohyeong;Lee, Seung Tae;Park, Choon-Keun;Hyun, Sang-Hwan;Lee, Eunsong
    • Journal of Embryo Transfer
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    • 제32권3호
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    • pp.139-146
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    • 2017
  • The objective of this study was to determine the effect of fructose that was supplemented to a chemically defined in vitro maturation (IVM) medium on oocyte maturation and embryonic development after parthenogenesis in pigs. The base medium for in vitro maturation (IVM) was porcine zygote medium (PZM) that was supplemented with 0.05% (w/v) polyvinyl alcohol (PVA) or 10% (v/v) porcine follicular fluid (pFF). In the first experiment, when immature pig oocytes were matured in a chemically defined medium that was supplemented with 5.5 mM glucose or with 1.5, 3.0 and 5.5 mM fructose, 3.0 mM fructose resulted in a higher nuclear maturation (91.5%) than 1.5 and 5.5 mM fructose (81.9 and 81.9%, respectively) but showed a similar result with 5.5 mM glucose (94.2%). However, there was no significant differences among groups in the embryo cleavage (89.4-92.4%), blastocyst formation (37.5-41.1%), and mean cell number of blastocyst (30.8-34.2 cells). Fructose at the concentration of 3.0 mM (1.08 pixels/oocyte) resulted in a higher intra-oocyte glutathione (GSH) content than 1.5 and 5.5 mM fructose (1.00 and 0.87 pixels/oocytes, respectively) while the cumulus cell expansion was not influenced. In the second experiment, effect of individual and combined supplementation of a chemically defined maturation medium with 5.5 mM glucose or 3.0 mM fructose was examined. No significant effect was found in the nuclear maturation (86.3-92.6%). Embryo cleavage was significantly increased by the combined supplementation with glucose and fructose (95.2%) compared to that with 3.0 mM fructose only (85.7%) while blastocyst formation (37.3-42.8%) and embryonic cell number (33.3-34.1 cells) were not altered. Effect of supplementation of pFF-containing medium with glucose and fructose + glucose was examined in the third experiment. No significant effect by the supplementation with glucose and fructose or glucose alone was observed in the nuclear maturation of oocytes (90.7-94.1%) and blastocyst formation (51.0-56.5%). Our results demonstrate that 3.0 mM fructose was comparable to 5.5 mM glucose in supporting in vitro oocyte maturation and embryonic development after parthenogenesis and could be used as an alternative energy source to glucose for in vitro maturation of pig oocytes.

Mechanism of Growth Inhibition by BCH in HEp2 Human Head and Neck Squamous Cell Carcinoma (사람 두경부 편평세포암종 HEp2 세포에서 BCH에 의한 세포성장 억제기전)

  • Choi, Bong-Kyu;Jung, Kyu-Yong;Cho, Seon-Ho;Kim, Chun-Sung;Kim, Do-Kyung
    • Journal of the Korean Society of Food Science and Nutrition
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    • 제37권5호
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    • pp.555-560
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    • 2008
  • Amino acid transporters are essential for the growth and proliferation in all living cells. Among the amino acid transporters, the system L amino acid transporters are the major nutrient transport system responsible for the $Na^+$-independent transport of neutral amino acids including several essential amino acids. The L-type amino acid transporter 1 (LAT1), an isoform of system L amino acid transporter, is highly expressed in cancer cells to support their continuous growth and proliferation. 2-Aminobicyclo-(2,2,1)-heptane-2-carboxylic acid (BCH) is a model compound for the study of amino acid transporter as a system L selective inhibitor. We have examined the effect and mechanism of BCH on cell growth suppression in HEp2 human head and neck squamous cell carcinoma. The BCH inhibited the L-leucine transport in a concentration-dependent manner with a $IC_{50}$ value of $51.2{\pm}3.8{\mu}M$ in HEp2 cells. The growth of HEp2 cells was inhibited by BCH in the timeand concentration-dependent manners. The formation of DNA ladder was not observed with BCH treatment in the cells. Furthermore, the proteolytic processing of caspase-3 and caspase-7 in the cells were not detected by BCH treatment. These results suggest that the BCH inhibits the growth of HEp2 human head and neck squamous cell carcinoma through the intracellular depletion of neutral amino acids for cell growth without apoptotic processing.

Activation of Bovine Oocytes by Combined Treatment with Ionomycin and cdc2 Kinase Inhibitor

  • Yoo, J.G.;Cho, S.R.;Lee, S.L.;Ock, S.A.;Rho, G.J.;Son, D.S.;Lee, H.J.;Choe, S.Y.
    • Journal of Embryo Transfer
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    • 제16권3호
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    • pp.223-231
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    • 2001
  • The success of nuclear transplantation with mammalian oocytes depends critically on the potential of oocytes activation, which mainly caused to prevent the re-accumulation of maturation promoting factor (MPF). This study was conducted to compare the effect of combined treatment of lonomycin with a Hl-histone kinase inhibitor (dimethylaminopurine, DMAP) or cdc2 kinase inhibitor (sodium pyrophosphate, SPP) on activation of bovine oocytes. In vitro matured bovine oocytes with the first polar body (PB) and dense cytoplasm were assigned to 3 experimental groups. For activation treatment, oocytcs were exposed to 5 $\mu$M lonomycin for 5 min (Group 1), and followed by 1.9 mM dimethylaminopurine (DMAP) for 3 h (Group 2) or followed by 2 mM sodium pyrophosphate (SPP) for 3 h (Group 3). The activation effects in the three treatments and the control group (untreated) were judged by the extrusion of the second PB and formation of a pronucleus (PN). Differences among groups were analysed using one-way ANOVA after arc-sine transformation of proportional data. All three treatments led to high activation rates (90% to 95%), with significant difference from the control. However, the extrusion of the second PB and the rate of PN formation differed remarkably among treatments. In Group I and 3, about 95% of the oocytes had extruded the second polar body, but one PN had formed in a higher proportion of oocytes in Group 3 than in Group 1 (90% vs. 5%). In experiment 2, the rates of cleavage and development into blastocysts in Group 1 were significantly lower than those of Group 2 and 3 (8.7% and 0% vs. 50.5% and 11.6%, and 44.6% and 7.2%, respectively, P<0.05). In experiment 3, ~80% of parthenotes in Group 1 were developed with haploid chromosomal sets. However, when ionomycin was followed immediately by DMAP (Group 2). only 20% of parthenotes were haploid. In Group 3, combined treatment with ionomycin and SPP, the appearance of abnormal chromosomal tracts was significantly (P〈0.05) reduced and the proportion of haploid parthenotes was increased to 85% (17/20) than in Group 2. These results demonstrate that SPP acted as a cdc2 kinase inhibitor and formed the haploidy in oocyte activation. Thus, the present study suggests that cdc2 kinase inhibitor, such as sodium pyrophosphate, may have an effective role in oocyte activation for the production of cloned embryos/animals by nuclear transplantation.

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Efficient Macrocyclization for Cyclicpeptide Using Solid-Phase Reaction

  • Kim, Joong-Hup;Hong, Il-Khee;Kim, Hyo-Jeong;Jeong, Hyeh-Jean;Choi, Moon-Jeong;Yoon, Chang-No;Jeong, Jin-Hyun
    • Archives of Pharmacal Research
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    • 제25권6호
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    • pp.801-806
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    • 2002
  • Cyclicpeptides are important targets in peptide synthesis because of their interesting biological properties. Constraining highly flexible linear peptides by cyclization is one of the mostly widely used approaches to define the bioactive conformation of peptides. Cyclic peptides often have increased receptor affinity and metabolic stability over their linear counterparts. We carried out virtual screening experiment via docking in order to understand the interaction between HLE-Human Leukocyte Elastase and ligand peptide and to identify the sequence that can be a target in various ligand peptides. We made cyclic peptides as a target base on Metlle-Phe sequence having affinity for ligand and receptor active site docking. There are three ways to cyclize certain sequences of amino acids such as Met-lie-Phe-Gly-Ile. First is head-to-tail cyclization method, linking between N-terminal and C-terminal. Second method utilizes amino acid side chain such as thiol functional group in Cys, making a thioether bond. The last one includes an application of resin-substituted amino acids in solid phase reaction. Among the three methods, solid phase reaction showed the greatest yield. Macrocyclization of Fmoc-Met-Ile-Phe-Gly-Ile-OBn after cleavage of Fmoc protection in solution phase was carried out to give macrocyclic compound 5 in about 7% yield. In the contrast with solution phase reaction, solid phase reaction for macrocyclization of Met-Ile-Phe-Gly-Ile-Asp-Tentagel in normal concentrated condition gave macrocyclic compound 7 in more than 35% yield.

Cloning and Expression of pcbAB Genes from Pseudomonas sp. DJ-12 in Escherichia coli (Pseudomonas sp. DJ-12 pcbAB 유전자의 Escherichia coli에서의 클로닝 및 발현)

  • 한재진;성태경;김치경
    • Korean Journal of Microbiology
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    • 제31권2호
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    • pp.129-134
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    • 1993
  • The pchAB genes of Pseudomonas sp. DJ-12 produce the enzymes of 4-chlorobipheny] (4CB) dioxygenase and dihydrodiol dehydrogenase which act on the first and second steps in degradation of 4CB and biphenyl. The genes were cloned in E coli XLI-Blue. The pcbAB genes of about 2.2 kb in size were contained in the pCUlO1 hybrid plasmid in the cloned cell of CUIOI. The genes were found to have their own promoter and three restriction sites for HindlII. 2,3-dihydroxybiphenyl was detected by the resting cell assay, as the metabolite transformed from biphenyl by the cloned cell of CUIOI. This means that the pcbAB genes are well expressed in E. coli. But dechlorination was unlikely involved in the pchAB gene expression but was believed to occur by functioning on 4CBA produced after ring-cleavage of 4CB.

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Expression, Purification and NMR Studies on MC4R-TM2 Mutant

  • Oh, Dae-Seok;Yun, Ji-Hye;Lee, Weon-Tae
    • Journal of the Korean Magnetic Resonance Society
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    • 제16권1호
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    • pp.34-45
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    • 2012
  • Melanocortin-4 receptor (MC4R) subtype is associated with obese humans. Especially, in a patient with severe early-onset obesity, novel heterozygous mutation in the MC4R gene was detected, resulting in an exchange of aspartic acid to asparagine in $90^{th}$ amino acid residue located in the predicted second trans-membrane domain (TM2). Mutations in the melanocortin-4 receptor (MC4R) gene are the most frequent monogenic causes of severe obesity which have been described as heterozygous with loss of function. In order to compare structure difference between MC4R wild type (MC4R-TM2-wt) and mutant (MC4R-TM2-D90N), we designed both MC4R-TM2-wt and MC4R-TM2-D90N construct in pET 21b vector. In this study, we optimized high-yield purification procedure for recombinant TM2-D90N. Eluted recombinant protein was resolubilized under urea condition for thrombin cleavage reaction and we conducted the high-performance liquid chromatography (HPLC) with reverse phase column under 1% acetonitrile, 0.01% TFA buffer solution. The molecular size of purified target peptide was confirmed by Tricine-SDS page analysis. To characterize MC4R-TM2-D90N, we have performed $^{15}N$-isotope labeling of peptide using M9 media and purified labeled target peptide for hetero-nuclear NMR spectroscopy.

Analysis of Social Distance using Election and Marriage Preference Data (사회적 거리분석: 선거자료와 결혼선호도자료에서 나타난 지역주의를 중심으로)

  • 이명진
    • Proceedings of the Korean Association for Survey Research Conference
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    • 한국조사연구학회 2001년도 춘계학술대회
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    • pp.37-57
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    • 2001
  • Much effort has been made to measure regional difference showing the pattern of regionalism in Korea. Focusing the concept of social distance, this paper examines various election and regional preference data. The paper presets three major findings. First current election data show clear regional cleavage. Second, regional discrimination and prejudice is often found in one' s everyday life. Third, political aspect of regionalism is strongly related to social aspect of regionalism. These findings suggest that regionalism in Korea will survive for the time being. The regionalism is rooted in everyday life. To political elites, it became an attractive and economic way to maintain their power. This tendency will promote the preference for parliamentary system among political elites and the possibility of appearance of regional party.