• Title/Summary/Keyword: Cytochrome $c_L$

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Apoptotic Effect of Rubia cordifolia Dichloromethane Extracts on Human Acute Jurkat T Cells (천초근 dichloromethane 추출물의 Jurkat T 세포에서 세포사멸 효과)

  • Kim, Ji-Hye;Lee, Jong-Hwan;Kim, Young-Ho;Kim, Kwang-Hyeon
    • Journal of Life Science
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    • v.19 no.2
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    • pp.163-168
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    • 2009
  • To understand cytotoxic activity of Rubia cordifolia L. (Rubiaceae), which has been used as a traditional oriental medicine, the mechanism underlying cytotoxic effect of its extract on human acute Jurkat T cells was investigated. The methanol extract of roots (3 kg) of R. codifolia was evaporated, dissolved in water, and then extracted by dichloromethane. The substances in the chloroform extract showing the most cytotoxic activity were further purified by a series of preparative HPLC. The extracted active substance (65 mg) was designated as CCH1. When Jurkat T cells were treated with CCH1 at concentration ranging from 0.5 to 2.0 ${\mu}g$/ml, apoptotic phenomena of cells companying several subsequent biochemical reactions such as mitochondria cytochrome c release, activation of casapase-8, -9, and caspase- 3, degradation of PARP and DNA fragmentation occurred via mitochondria-dependent pathway. However, abrogation of apoptosis was observed in an ectopic expression of Bcl-xL, which is a suppressor for mitochondrial cytochrome c release. These results demonstrate that the cytotoxicity of CCH1 against Jurkat T cells is attributable to apoptosis mediated by mitochodria-dependent death-signaling regulated by Bcl-xL. In addition, the CCH1 is more potent to leukemia Jurkat T cell than to human peripheral blood monocyte cells (PBMC).

Effect of Low Temperature upon the Fatty Acid Composition Plasma Membrane of Canola (저온 환경이 Canola 원형질막의 Fatty Acid 구성에 미치는 영향)

  • Kwon, Sung-Hwan;Plank, D.W.;Jeon, Hee;Kim, Jae-Chul
    • Journal of Bio-Environment Control
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    • v.4 no.2
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    • pp.136-143
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    • 1995
  • Using a PEG- dextran two phase partition method, plasma and intracellular membrane separated from microsomal membrane of canola (Brassica napus) leaves have been fractionated by centrifugation. $K^{+}$- ATPase specific activity in the plasma membrane (U$_2$ phase) of plants grown at $25^{\circ}C$ and 1$0^{\circ}C$ were 6.6 and 4.6 times, respectively that of the microsomal membrane. Plasma membrane had a lower cytochrome- c- oxidase specific activity than the microsomal membrane or intracellular membrane, while intracellular membrane (L$_2$ phase) had a high cytochrome-c- oxidase but little $K^{+}$- ATPase specific activity. The plasma membrane of canola grown at 1$0^{\circ}C$ had higher 18:3 to 18:2 (linolenic to linoleic acid) ratio (29.2% ) and higher degree of unsaturation than that grown at $25^{\circ}C$ The double bond index of plasma membrane from canola grown at 1$0^{\circ}C$ increased by 8.9% relative to canola grown at $25^{\circ}C$. Similar, intracellular membrane increased by 19.7% at 1$0^{\circ}C$. Canola grown at 1$0^{\circ}C$ was lower in chlorophyll contents (17.3%) than that grown at $25^{\circ}C$. These changes in fatty acid unsaturation were attributable largely to change in Cl8 fatty acid, with major changes occurring in linolenic acid (18 :3) which might have a physiological role of membrane to adaptation on low temperature.ure.

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Apoptotic Activity of Insect Pathogenic Fungus Paecilomycesc japonica Toward Human Acute Leukemia Jurkat T Cells is Associated with Mitochondria-Dependent Caspase-3 Activation Regulated by Bcl-2

  • Park, Hye-Won;Jen, Do-Youn;Kim, Young-Ho
    • Journal of Microbiology and Biotechnology
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    • v.12 no.6
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    • pp.950-956
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    • 2002
  • The antitumor activity of the insect pathogenic fungus Paecilomyces japonica has been attributed to apoptotic cell death. However, the mechanism underlying the induced apoptosis has not yet been elucidated. In this study, we for the first time show that mitochondria-dependent caspase-3 activation were associated with the apoptotic activity of P. japonica in human acute leukemia Jurkat T cells. When Jurkat T cells were treated with the ethyl acetate extract of P japonica at concentrations ranging from $2-6{\mu}g/ml$, apoptotic cell death. accompanied by several biochemical events such as caspase-9 activation, caspase-3 activation, degradation of poly (ADP-ribose) polymerase (PARP), and apoptotic DNA fragmentation, was induced in a dose-dependent manner. In addition, the release of cytochrome c from mitochondria was detected. Under these conditions, the expression of Fas and Fas-ligand (FasL) remained unchanged. Ethyl acetate extract-induced mitochondrial cytochrome c release, caspase-3 activation, PARP cleavage, and apoptotic DNA fragmentation were suppressed by the ectopic expression of Bcl-2, which is known to block mitochondrial cytochrorme c release. Accordingly, these results demonstrate that P. japonica-induced apoptotic cell death is mediated by a cytochrome c-dependent caspase-3 activation pathway that can be interrupted by Bcl-2.

Effect of Mutations of Five Conserved Histidine Residues in the Catalytic Subunit of the cbb3 Cytochrome c Oxidase on its Function

  • Oh Jeong-Il
    • Journal of Microbiology
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    • v.44 no.3
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    • pp.284-292
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    • 2006
  • The cbb3 cytochrome c oxidase has the dual function as a terminal oxidase and oxygen sensor in the photosynthetic bacterium, Rhodobacter sphaeroides. The cbb3 oxidase forms a signal transduction pathway together with the PrrBA two-component system that controls photosynthesis gene expression in response to changes in oxygen tension in the environment. Under aerobic conditions the cbb3 oxidase generates an inhibitory signal, which shifts the equilibrium of PrrB kinase/phosphatase activities towards the phosphatase mode. Photosynthesis genes are thereby turned off under aerobic conditions. The catalytic subunit (CcoN) of the R. sphaeroides cbb3 oxidase contains five histidine residues (H2l4, B233, H303, H320, and H444) that are conserved in all CcoN subunits of the cbb3 oxidase, but not in the catalytic subunits of other members of copper-heme superfamily oxidases. H214A mutation of CcoN affected neither catalytic activity nor sensory (signaling) function of the cbb3 oxidase, whereas H320A mutation led to almost complete loss of both catalytic activity and sensory function of the cbb3 oxidase. H233V and H444A mutations brought about the partial loss of catalytic activity and sensory function of the cbb3 oxidase. Interestingly, the H303A mutant form of the cbb3 oxidase retains the catalytic function as a cytochrome c oxidase as compared to the wild-type oxidase, while it is defective in signaling function as an oxygen sensor. H303 appears to be implicated in either signal sensing or generation of the inhibitory signal to the PrrBA two-component system.

In vitro inhibitory effects of Yangguksanhwa-tang and Taeumjowi-tang on CYP450 isozymes (양격산화탕과 태음조위탕 추출물의 in vitro CYP450 효소 활성 억제 효과)

  • Ha, Hye-Kyung;Jin, Seong-Eun;Shin, Hyeun-Kyoo
    • Journal of Society of Preventive Korean Medicine
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    • v.16 no.3
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    • pp.129-137
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    • 2012
  • Objective : Herb-drug interactions have become an important issue because of the consumption of herbal remedies has increased in the world. Yangguksanhaw-tang (Liang ge san huo-tang) and Taeumjowi-tang (Tai yin tiao wei-tang) are typical herbal formulas on Sasang constitution medicine (four-constitution medicine). This study was aimed at evaluating the effects of Yangguksanhaw-tang and Taeumjowi-tang on drug metabolizing enzymes, cytochrome P450 (CYP450) isozymes. Methods : Vivid$^{(R)}$ CYP450 Screening Kits were used to measure of CYP3A4, CYP2C19, CYP2D6 and CYP2E1 activities. This method is based on the use of fluorescent CYP450 substrates that are efficiently metabolized by specific CYP450 isozymes to yield a product with altered fluorescent properties. The percent inhibitions of CYP450s by herbal formulas were calculated. Results : Yangguksanhaw-tang inhibited CYP2C19 and CYP2E1 activities higher than that other CYP450 isozymes. The $IC_{50}$ values of CYP2C19 and CYP2E1 were 159.83 ${\mu}g/mL$ and 261.40 ${\mu}g/mL$, respectively. The CYP2E1 activity was inhibited ($IC_{50}=215.17{\mu}g/mL$) higher than that other CYP450 isozymes by Taeumjowi-tang. Conclusions : These results suggest that Yangguksanhaw-tang may inhibit the metabolism of co-administered drugs whose primary route of metabolism is via CYP2C19 or CYP2E1. Taeumjowi-tang could inhibit the metabolism of co-administered drugs, which are substrates for CYP2E1. Therefore, co-administration of the herbal formulas and other conventional drugs should be undertaken with care.

Homology Modeling and In Vitro Analysis for Characterization of Streptomyces peucetius CYP157C4

  • Rimal, Hemraj;Yu, Sang-Cheol;Jang, Jong Hwa;Oh, Tae-Jin
    • Journal of Microbiology and Biotechnology
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    • v.25 no.9
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    • pp.1417-1424
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    • 2015
  • In this study, we tried to characterize Streptomyces peucetius CYP157C4 with homology modeling using three cytochrome P450 (CYP) structures (CYP157C1, CYP164A2, and CYP107L1), having discovered that CYP157C4 lacks the ExxR motif that was considered invariant in all CYPs. We used Discovery Studio 3.5 to build our model after first assessing the stereochemical quality and side-chain environment, and a 7-ethoxycoumarin substrate was docked into the final model. The model-substrate complex allowed us to identify functionally important residues and validate the active-site architecture. We found a distance of 4.56 Å between the 7-ethoxycoumarin and the active site of the heme, and cloning and an in vitro assay of the CYP157C4 showed the dealkylation of the substrate. Since the details regarding this group of CYP structures are still unknown, the findings of this study may provide elucidation to assist with future efforts to find a legitimate substrate.

Distribution and Bionomics of the Argentine Ant Linepithema humile (Mayr) (Hymenoptera: Formicidae: Dolichoderinae)

  • Min-Ji Lee;Young-Gyu Ban;Heejo Lee;Young Ha Kim;Dayeong Kim;Nang-Hee Kim;Dong Eon, Kim
    • Proceedings of the National Institute of Ecology of the Republic of Korea
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    • v.4 no.3
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    • pp.104-114
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    • 2023
  • In this study, the distribution, bait preference, hosts, symbionts, habitat status, and cytochrome c oxidase subunit-I sequences of the ant Linepithema humile were analyzed. This species habitat comprised approximately 1.5 km along stone crevices, flower gardens, roadsides, and container yards in the vicinity of Busan station, Korea. Feeding activity of this species was confirmed in 9 species of 7 families of plants. Which was approximately 14% of total flora (63 species of 37 families) in the studied area. Particularly, it was observed that Dendranthema indicum and Camellia japonica were the most frequently visited. Further, we verified that they interacted with four species of aphids. We identified 22 species of ants (17 genera, 4 subfamilies) inhabiting the Busan station area, including L. humile, Tetramorium tsushimae, Nylanderia flavipes and the alien species Paratrechina longicornis. In areas where L. humile was the dominant species, the habitat of local native ants was reduced by approximately 30%. Bait preference was ranked in the following order: jelly, water with glucose (25%), and yogurt, with the highest preference for baits containing sugar and protein. The 630-bp cytochrome c oxidase subunit-I sequences of the local L. humile populations were 99% identical to those of L. humile in the National Center for Biotechnology Information database, and the Argentine ants that invaded Korea had a sequence identical to that of other invasive populations from China, Japan, and New Zealand. As L. humile rapidly increases due to domestic adaptation, continuous monitoring and control strategy for eradication are needed to protect domestic biodiversity.

Inhibition of Cytochrome P450 by Propolis in Human Liver Microsomes

  • Ryu, Chang Seon;Oh, Soo Jin;Oh, Jung Min;Lee, Ji-Yoon;Lee, Sang Yoon;Chae, Jung-woo;Kwon, Kwang-il;Kim, Sang Kyum
    • Toxicological Research
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    • v.32 no.3
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    • pp.207-213
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    • 2016
  • Although propolis is one of the most popular functional foods for human health, there have been no comprehensive studies of herb-drug interactions through cytochrome P450 (CYP) inhibition. The purpose of this study was to determine the inhibitory effects of propolis on the activities of CYP1A2, 2A6, 2B6, 2C9, 2C19, 2D6, 2E1 and 3A4 using pooled human liver microsomes (HLMs). Propolis inhibited CYP1A2, CYP2E1 and CYP2C19 with an $IC_{50}$ value of 6.9, 16.8, and $43.1{\mu}g/mL$, respectively, whereas CYP2A6, 2B6, 2C9, 2D6, and 3A4 were unaffected. Based on half-maximal inhibitory concentration shifts between microsomes incubated with and without nicotinamide adenine dinucleotide phosphate, propolis-induced CYP1A2, CYP2C19, and CYP2E1 inhibition was metabolism-independent. To evaluate the interaction potential between propolis and therapeutic drugs, the effects of propolis on metabolism of duloxetine, a serotonin-norepinephrine reuptake inhibitor, were determined in HLMs. CYP1A2 and CYP2D6 are involved in hydroxylation of duloxetine to 4-hydroxy duloxetine, the major metabolite, which was decreased following propolis addition in HLMs. These results raise the possibility of interactions between propolis and therapeutic drugs metabolized by CYP1A2.