• Title/Summary/Keyword: cleavage products

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Trichomonas vaginalis Metalloproteinase Induces mTOR Cleavage of SiHa Cells

  • Quan, Juan-Hua;Choi, In-Wook;Yang, Jung-Bo;Zhou, Wei;Cha, Guang-Ho;Zhou, Yu;Ryu, Jae-Sook;Lee, Young-Ha
    • Parasites, Hosts and Diseases
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    • v.52 no.6
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    • pp.595-603
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    • 2014
  • Trichomonas vaginalis secretes a number of proteases which are suspected to be the cause of pathogenesis; however, little is understood how they manipulate host cells. The mammalian target of rapamycin (mTOR) regulates cell growth, cell proliferation, cell motility, cell survival, protein synthesis, and transcription. We detected various types of metalloproteinases including GP63 protein from T. vaginalis trophozoites, and T. vaginalis GP63 metalloproteinase was confirmed by sequencing and western blot. When SiHa cells were stimulated with live T. vaginalis, T. vaginalis excretory-secretory products (ESP) or T. vaginalis lysate, live T. vaginalis and T. vaginalis ESP induced the mTOR cleavage in both time-and parasite load-dependent manner, but T. vaginalis lysate did not. Pretreatment of T. vaginalis with a metalloproteinase inhibitor, 1,10-phenanthroline, completely disappeared the mTOR cleavage in SiHa cells. Collectively, T. vaginalis metallopeptidase induces host cell mTOR cleavage, which may be related to survival of the parasite.

Caspase-3-facilitated Stoichiometric Cleavage of a Large Recombinant Polyprotein (카스파제-3 효소를 이용한 폴리-단백질의 정량적 프로세싱 분석)

  • Kim, Moonil
    • Journal of Life Science
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    • v.25 no.4
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    • pp.385-389
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    • 2015
  • In this study, it is reported that a large polyprotein can be stoichiometrically cleaved by the use of caspase-3-dependent proteolysis. Previously, it has been shown that the proteolytic IETD motif was partially processed when treated with caspase-3, while the DEVD motif was completely cleaved. The cleavage efficiency of the DEVD-based substrate was approximately 2.0 times higher than that of the IETD substrate, in response to caspase-3. Based on this, 3 protein genes of interest were genetically linked to each other by adding two proteolytic cleavage sequences, DEVD and IETD, for caspase-3. Particularly, glutathione-S transferase (GST), maltose binding protein (MBP), and red fluorescent protein (RFP) were chosen as model proteins due to the variation in their size. The expressed polyprotein was purified by immobilized metal ion affinity chromatography (IMAC) via a hexa-histidine tag at the C-terminal end, showing 93 kDa of a chimeric GST:MBP:RFP fusion protein. In response to caspase-3, cleavage products, such as MBP:RFP (68 kDa), MBP (42 kDa), RFP (26 kDa), and GST (25 kDa), were separated from a large precursor GST:MBP:RFP (93 kDa) via SDS-PAGE. The results obtained from this study indicate that a multi-protein can be stoichiometrically produced from a large poly-protein by using proteolytic recognition motifs, such as DEVD and IETD tetra-peptides, for caspase-3.

A Study on the Character of Okchun Feldspar (옥천 장석의 특성에 관한 연구)

  • 최용식;이종근;이경희;이병하;김영호
    • Journal of the Korean Ceramic Society
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    • v.27 no.2
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    • pp.149-154
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    • 1990
  • Before benefication of Okchun feldspar, all kinds of Fe-compounds and mafic minerals which were included in feldspar were studied. Also to produce 30/140mesh products as glass materials, optimum grinding condition and degree of liberation were studied. Fe-compounds were Biotite and Limonite, and these were existed along the cleavage. Optimum grinding time was 2min. and at this time weight percent of 30/140mesh products was 72.7% and degree of liberation at this particle size was 91.5%.

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Identification of Homoserine Lactone Derivatives Using the Methionine Functionalized Solid Phase Synthesis by Gas Chromatography/Mass Spectrometry

  • Moon, Hong-Sik
    • Archives of Pharmacal Research
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    • v.27 no.1
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    • pp.25-30
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    • 2004
  • Combinatorial homoserine lactone mixtures and individual products were obtained from the methionine-functionalized resin in solid-phase synthesis. The four-step process consisting of a coupling step of an N-Fmoc-L-methionine, deprotection of N-Fmoc group, N-coupling with a carboxylic acid, and cleavage reaction through a polymer supported strategy is described. Gas chromatography-mass selective detector (GC-MSD) techniques provide the most powerful methods for identifying both the combinatorial mixtures and individual products.

Biodegradation of Kraft Lignins by White-Rot Fungi(I) -Lignin from Pitch Pine- (백색부후균에 의한 크라프트 리그닌의 분해(I) -리기다소나무 리그닌-)

  • 김명길;안원영
    • Journal of Korea Foresty Energy
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    • v.17 no.1
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    • pp.56-70
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    • 1998
  • This study was carried out to investigate the structural characteristics of kraft lignin and the wood degrading characteristics, the productivity of ligninolytic enzymes and the enzymatic degradation of kraft lignin by white-rot fungi. To purify kraft lignin, precipitation of kraft pulping black liquors of pitch pine meal was done by titration with lN $H_{2}SO_{4}$ reaching to pH 2, and isolation of the precipitates done by centrifugation. The isolated precipitates from pitch pine were redissloved in lN NaOH, reprecipitated by titration with lN $H_{2}SO_{4}$, washed with deionized water, and kept ofr analysis after freeze drying. Fractionation of the precipitates in solution by successive extraction with $CH_{2}Cl_{2}$ and MeOH, and the fractionates were named SwKL, SwKL I, SwKL II, and SwKL III for pitch pine kraft lignin. The more molecular weights of kraft lignin increased, the less phenolic hydroxyl groups and the more aliphatic hydroxyl groups. Because as the molecular weights increased, the ratio of etherified guaiayl/syringyl(G/S ratio) and the percentage were increased. The spectra obtained by 13C NMR and FTIR assigned by comparing the chemical shifts of various signals with shifts of signals from autherized ones reported. The optimal growth temperature and pH of white-rot fungi in medium were $28^{\circ}C$ and 4.5-5.0, respectively. Especially, in temperature and pH range, and mycelial growth, the best white-rot fungus selected was Phanerochaete chrysosporium for biodegradation. For the degradation pathways, the ligninolytic fungus jcultivated with stationary culture using medium of 1% kraft lignin as a substrate for 3 weeks at $28^{\circ}C$. The weight loss of pitch pine kraft lignin was 15.8%. The degraded products extracted successively methoanol, 90% dioxane and diethyl ether. The ether solubles were analyzed by HPLC. Kraft lignin degradation was initiated in $\beta$-O-4 bonds of lignin by the laccase from Phanerochaete chrysosporium and the degraded compounds were produced from the cleavage of $C\alpha$-$C\beta$ linkages at the side chains by oxidation process. After $C\alpha$-$C\beta$ cleavage, $C\alpha$-Carbon was oxidized and changed into aldehyde and acidic compounds such as syringic acid, syringic aldehyde and vanilline. And the other compound as quinonemethide, coumarin, was analyzed. The structural characteristics of kraft lignin were composed of guaiacyl group substituted functional OHs, methoxyl, and carbonyl at C-3, -4, and -5 and these groups were combinated with $\alpha$ aryl ether, $\beta$ aryl ether and biphenyl. Kraft lignin degradation pathways by Phanerochaete chrysosporium were initially accomplished cleavage of $C\alpha$-$C\beta$ linkages and $C\alpha$ oxidation at the propyl side chains and finally cleavage of aromatic ring and oxidation of OHs.

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Substituent Effect in Photochemistry of Carbonyl Compounds: α-Halovalerophenones

  • Cho, Sung-Su;Park, Bong-Ser
    • Bulletin of the Korean Chemical Society
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    • v.25 no.1
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    • pp.42-44
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    • 2004
  • Valerophenones containing a substituent at alpha position to the carbonyl group show the remarkable substituent effects on their photochemical reactions. ${\alpha}$-Bromovalerophenone gives only the C-Br bond cleavage products, but the ${\alpha}$-chlorovalerophenone follows the classical Norrish/Yang reaction pathway predominantly.

Reaction Pattern of Bacillus cereus D-11 Chitosanase on Chitooligosaccharide Alcohols

  • Gao, Xing-Ai;Jung, Woo-Jin;Kuk, Ju-Hee;Park, Ro-Dong
    • Journal of Microbiology and Biotechnology
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    • v.19 no.4
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    • pp.358-361
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    • 2009
  • The purified endochitosanase(Mw 41 kDa) from bacterium Bacillus cereus D-11 hydrolyzed chitooligomers $(GlcN)_{5-7}$ into chitobiose, chitotriose, and chitotetraose as the final products. The minimal size of the oligosaccharides for enzymatic hydrolysis was a pentamer. To further investigate the cleavage pattern of this enzyme, chitooligosaccharide alcohols were prepared as substrates and the end products of hydrolysis were analyzed by TLC and HPLC. The chitosanase split $(GlcN)_4GlcNOH$ into $(GlcN)_3+(GlcN)_1GlcNOH$, and $(GlcN)_5GIcNOH$ into $(GlcN)_4+(GlcN)_1GlcNOH$ and $(GlcN)_3+(GlcN)_2GlcNOH$. The heptamer $(GlcN)_6GlcNOH$ was split into $(GlcN)_5$ [thereafter hydrolyzed again into $(GlcN_3+(GlcN)2]+(GlcN)_1GlcNOH$, $(GlcN)_4+(GlcN)_2GlcNOH$, and $(GlcN)_3+(GlcN)_3GlcNOH$, whereas $(GlcN)_{1-3}GlcNOH$ was not hydrolyzed. The monomers GlcN and GIcNOH were never detected from the enzyme reaction. These results suggest that D-11 chitosanase recognizes three glucosamine residues in the minus position and simultaneously two residues in the plus position from the cleavage point.

Density Functional Studies of Ring-Opening Reactions of Li+-(ethylene carbonate) and Li+-(vinylene carbonate)

  • Han, Young-Kyu;Lee, Sang-Uck
    • Bulletin of the Korean Chemical Society
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    • v.26 no.1
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    • pp.43-46
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    • 2005
  • Reaction energies were determined for reductive ring-opening reactions of Li$^+$-coordinated ethylene carbonate (EC) and vinylene carbonate (VC) by a density functional method. We have also explored the ring-opening of Li$^+$-EC and Li$^+$-VC by reaction with a nucleophile (CH$_3$O$^-$.) thermodynamically. Our thermodynamic calculations led us to conclude that the possible reaction products are CH$_3$OCH$_2$CH$_2$OCO$_2$Li (O$_2$-C$_3$ cleavage) for Li$^+$-EC +CH$_3$O$^-$., and CH$_3$OCHCHOCO$_2$Li (O$_2$-C$_3$ cleavage) and CH$_3$OCO$_2$CHCHOLi (C$_1$-O$_2$ cleavage) for Li$^+$-VC +CH$_3$O$^-$.. The opening of VC would occur at the C$_1$-O$_2$ side by a kinetic reason, although the opening at the O$_2$-C$_3$ side is more favorable thermodynamically.

Caspase Cleavage of Receptor Tyrosine Kinases in the Dependence Receptor Family

  • Gyu Hwan Park;Yoo Kyung Kang;Seung-Mann Paek;Chan Young Shin;Sun-Young Han
    • Biomolecules & Therapeutics
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    • v.31 no.4
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    • pp.359-369
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    • 2023
  • Dependence receptors are a group of receptor proteins with shared characteristics of transducing two different signals within cells. They can transduce a positive signal of survival and differentiation in the presence of ligands. On the other hand, dependence receptors can transduce an apoptosis signal in the absence of ligands. The function of these receptors depends on the availability of their ligands. Several receptor tyrosine kinases (RTKs) have been reported as dependence receptors. When cells undergo apoptosis by dependence receptors, the intracellular domain of some RTKs is cleaved by the caspases. Among the RTKs that belong to dependence receptors, we focused on eight RTKs (RET, HER2, MET, ALK, TrkC, EphA4, EphB3, and c-KIT) that are cleaved by caspases. In this review, we describe the features of the receptors, their cleavage sites, and the fate of the cleaved products, as well as recent implications on them being used as potential therapeutics for cancer treatment.

Wood Characteristics of Phyllastachys bambusoides, Phyllostachys nigra var. henonis, and Phyllastachys pubescens Grown in Damyang District (담양지역(潭陽地域) 왕대, 분죽 및 맹종죽의 재질특성(材質特性))

  • So, Won-Tek;Kim, Yoon-Soo;Chung, Woo-Yang;Lee, Hyoung-Woo
    • Journal of the Korean Wood Science and Technology
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    • v.27 no.2
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    • pp.7-14
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    • 1999
  • This study was performed to investigate the basic wood characteristics of Phyllostachys bambusoides, Phyllostaehys nigra var. henonis, and Phyllostaehys pubeseens grown in Damyang district in Korea. There were no significant differences in the anatomical structure of three species and they showed rediculate vessels and tylosis. The bulk density of P. bambusoides, P. nigra var. henonis, and P. pubeseens was 0.61g/$cm^3$, 0.66g/$cm^3$, and 0.58g/$cm^3$ P. respectively. P. pubescens showed the highest cleavage resistance and P. nigra var. henonis showed the lowest cleavage resistance, and therefore P. nigra var. henonis was fittest for the cleaving prosessing of bamboo.

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