• Title/Summary/Keyword: cytochrome $c_3$

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Photoinduced Electron Tansfer of Microperoxidase-8

  • Su, Chien-Hua;Lin, Ren-Jay;Chang, I-Jy
    • Journal of Photoscience
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    • v.6 no.3
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    • pp.129-133
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    • 1999
  • Microperoxidase 8 (MP8) has been prepared by sequential hydrolysis of cytochrome C by pepsin and trypsin. This five-coordinated heme-octapetide fragment provides a uniques structure to evaluate the electronic coupling efficiency to the iron through axial position and porphyrin edge. At alkali pH, Ru(bpy)2(im)22+ is completely quenched in AcMP8Ru complex . Transient kinetics measurement showed the decay rate to be ~1 $\times$1012S-1. Ruthenium bipyridine complex with a carboxyl group substituted bipyridine has been prepared adn reacted with MR 8 to yield N-terminus bound RuMP8 complex. The luminescence decay rate has been measured as 1 $\times$109S-1. By using semiclassical electron transfer theory, we found the electron transfer efficiency through axial positioin of iron prophyrin is as good through prophyrin edge.

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First Reliable Record of the Keeltail Pomfret Taractes rubescens (Bramidae: Perciformes) from Korea (한국산 새다래과(Bramidae) 어류 1미기록종, Taractes rubescens (Jordan and Evermann, 1887))

  • Lee, Jae-Hwan;Lee, Woo-Jun;Kim, Jin-Koo
    • Korean Journal of Fisheries and Aquatic Sciences
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    • v.52 no.3
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    • pp.283-287
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    • 2019
  • Two specimens of Taractes rubescens (family Bramidea) were collected from Busan and Pohang, Korea, between 2015 and 2016. Taractes rubescens was very similar to Taractes asper, but the two species are clearly distinguishable by the number of anal fin rays (21-23 in T. rubescens vs. 25-27 in T. asper) and the presence of a lateral caudal keel (present in T. rubescens vs. absent in T. asper). The mitochondrial DNA cytochrome c oxidase subunit I sequences of the specimens described in this paper perfectly matched those of T. rubescens, but differed from those of T. asper (genetic distance: 6.9%). We use the Korean common name proposed by Kim and Ryu (2017) for T. rubescens, "Geom-eun-sae-da-rae".

DNA Barcoding for the Hydrothermal Vent Crab Austinograea Species (Crustacea: Bythograeidae) from the North Fiji Basin, Southwestern Pacific Ocean

  • Lee, Won-Kyung;Ju, Se-Jong;Hou, Bo Kyeng;Kim, Se-Joo
    • Animal Systematics, Evolution and Diversity
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    • v.35 no.1
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    • pp.30-32
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    • 2019
  • The brachyuran crab Bythograeidae Williams, 1980 is common in hydrothermal vent fields worldwide and has recorded to sixteen species of six genera. In this study, we firstly determined the cytochrome c oxidase subunit 1 (CO1) DNA barcodes for the fifth species of Austinograea, A. hourdezi, from hydrothermal vent regions of the North Fiji Basin in southwestern Pacific Ocean. All CO1 DNA barcodes of A. hourdezi were identical. The interspecies variations of three bythograeid genera were 10.9-13.3% for Austinograea, 6.6-15.7% for Bythograea, and 9.7% for Gandalfus. These results would be helpful to understand taxonomy of brachyuran crabs living in hydrothermal vent fields using CO1 DNA barcodes.

Induction of Apoptosis by N-nitrosocarbofuran, via Cytochrome c-Mediated Activation of Caspase-3 protease

  • Lee, Bang-Wool;Oh, Seon-Hee;Lee, Byung-Hoon
    • Proceedings of the PSK Conference
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    • 2003.04a
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    • pp.158.1-158.1
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    • 2003
  • Carbofuran(CF) is one of the most widely used carbamate pesticides in the world applied for insect and nematode control. Due to its widespread use in agriculture and households, contamination of food, water, and air has become serious, and consequently adverse health effects are inevitable in humans, animals, wildlife and fish, it has reported that CF alone or in combination with other carbamate insecticides influences the level of reproductive and metabolic hormones such as thyroxine and corticosterone, and results in impairment of endocrine, immune behavioral functions. (omitted)

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DNA Barcoding of Aegista chejuensis and Plectotropis quelpartensis (Gastropoda: Stylommatophora: Camaenidae)

  • Kang-San Kim;Jun-Sang Lee
    • Animal Systematics, Evolution and Diversity
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    • v.39 no.4
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    • pp.295-299
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    • 2023
  • Two land snails, Aegista chejuensis (Pilsbry and Hirase, 1908) and Plectotropis quelpartensis (Pilsbry and Hirase, 1908), are endemic to Korea and were collected from Hataedo and Jodo Islands in the Yellow Sea of South Korea, respectively. Many terrestrial snail habitats have been confirmed in Korea; however, their genetic sequences have rarely been reported. This study describes the mitochondrial cytochrome c oxidase subunit I gene (COI) sequences of two species, followed by an analysis of the genetic distance between these two species and their congeners. As a result, there was no intra-species variation in both species A. chejuensis or P. quelpartensis. However, the inter-species variation was clear (10.3-31.5%). We provide photographs and a brief diagnosis for morphological verification.

Induction of Apoptosis by (-)-epigallocatechin-3-gallate in HL-60 Cells (인체 혈액암세포주(HL-60)에서 (-)-epigallocatechin-3-gallate에 의한 Aapoptosis 유도)

  • 이해미;김연정;박태선
    • Journal of Nutrition and Health
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    • v.36 no.4
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    • pp.382-388
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    • 2003
  • (-)-Epigallocatechin-3-gallate (EGCG) is a polyphenolic compound found in peen tea leaves, and has been known to be one of the most potent catechin species which inhibits cell growth most possibly through an apoptotic cell death. We investigated the apoptotic activity of (-)-EGCG on the human myeloid leukemia cell line, HL-60. Our results of MTT test indicated that (-)-EGCG had a significant antiproliferation effect in HL-60 cells with $IC_{50}$/ (50% inhibition concentration) value of 65 $\mu$M. Giemsa statining of HL-60 cells treated with (-)-EGCG (100 $\mu$M) for 6hrs showed a typical apoptosis-specific morphological change including shrinkage of the cytoplasm, membrane blobbing and compaction of the nuclear chromatin. The DNA fragmentation was observed from the agarose gel electrophoresis of cells treated with (-)-EGCG for 3hrs or longer, and was progressed to a greater degree as treatment time increases. Treatment of the cells with (-)-EGCG (100 $\mu$M) resulted in a rapid release of mitochondrial cytochrome c into the cytosol, and a subsequent cleavage of caspase-3 to an active form in a treatment-time dependent manner. (-)-EGCG (100 $\mu$M) also stimulated proteolytic cleavage of poly-(ADP-ribose) polymerase (PARP) to an active form in HL-60 cells. Tlken together, (-)-EGCG appears to induce the apoptosis in human myeloid leukemia cells via a caspase-dependent pathway. These results suggest the possible application of (-)-EGCG, the major active compound in green tea, as an antiproliferative agent for cancer prevention.

Application for Identification of Food Raw Materials by PCR using Universal Primer (일반 프라이머를 이용한 PCR의 식품원료 진위 판별에 적용)

  • Park, Yong-Chjun;Jin, Sang-Ook;Lim, Ji-Young;Kim, Kyu-Heon;Lee, Jae-Hwang;Cho, Tae-Yong;Lee, Hwa-Jung;Han, Sang-Bae;Lee, Sang-Jae;Lee, Kwang-Ho;Yoon, Hae-Seong
    • Journal of Food Hygiene and Safety
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    • v.27 no.3
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    • pp.317-324
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    • 2012
  • In order to determine an authenticity of food ingredient, we used DNA barcode method by universal primers. For identification of animal food ingredients, LCO1490/HCO2198 and VF2/FISH R2 designed for amplifying cytochrome c oxidase subunit1 (CO1) region and L14724/H15915 for cytochrome b (cyt b) region on mitochondrial DNA were used. Livestock (cow, pig, goat, sheep, a horse and deer) was amplified by LCO1490/HCO 2198, VF2/FISH R2 and L14724/H15915 primers. Poultry (chicken, duck, turkey and ostrich) was amplified by LCO1490/HCO 2198 and VF2/FISH R2 primers. But, Fishes (walleye pollack, herring, codfish, blue codfish, trout, tuna and rockfish) were only amplified by VF2/FISH R2 primers. For plant food ingredients, 3 types of primers (trnH/psbA, rpoB 1F/4R and rbcL 1F/724R) have been used an intergenic spacer, a RNA polymerase beta subunit and a ribulose bisphosphate carboxylase region on plastid, respectively. Garlic, onion, radish, green tea and spinach were amplified by trnH/psbA, rpoB 1F/4R and rbcL 1F/724R. The PCR product sizes were same by rpoB 1F/4R and rbcL 1F/724R but, the PCR product size using trnH/psbA primer was different with others for plants each. We established PCR condition and universal primer selection for 17 item's raw materials for foods and determine base sequences aim to PCR products in this study. This study can apply to determine an authenticity of foods through making an comparison between databases and base sequences in gene bank. Therefore, DNA barcode method using universal primers can be a useful for species identification techniques not only raw materials but also processed foods that are difficult to analyze by chemical analysis.

Apoptosis Suppressor에 관련된 유전자 스크린 방법과 동정된 유전자 특성 규명

  • 황규찬;옥도원;권득남;신혜경;김진회
    • Proceedings of the KSAR Conference
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    • 2001.03a
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    • pp.16-16
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    • 2001
  • Apoptosis로 일컬어지는 예정된 세포사멸(programmed cell death)은 개별 세포의 입장에서는 곧바로 사멸을 의미하지만, 정상적인 고등 생물의 입장에서는 개체의 발생과 분화하는데 프로그램된 과정이다. 자발적 세포사멸은 다른 조직에 비해 생식 조직인 난소나 정소에서 복잡한 apoptosis 기작들을 가지리라 사료된다. 본 연구는 Bcl-2 family중 apoptotic protein인 Bax에 대해 suppression하는 유전자를 yeast system을 활용하여 돼지 정소와 난소로부터 각각 cDNA library를 구축한 후 탐색하였다. 탐색에 활용된 cDNA library는 돼지의 정소와 난소로부터 mRNA를 분리하여 yeast vector인 pAD-GAL4-2.1에 구축하였고, 마우스 bax 유전자는 gal 1 promoter의 조절 하에 glucose 배지에서는 유도되지 않고, galactose 배지에서만 선택적으로 Bax를 발현할 수 있는 효모 vector(pL19-bax)를 구축하였다. Bax에 의한 apoptosis suppressor를 탐색하기 위해 우선 효모 W303에 pL19-bax를 transform하여 glucose 배지에서 Bax의 발현을 억제하였다. pL19-bax를 가진 효모에 정소와 난소로부터 구축된 cDNA library를 transform 시키고, transform된 효모는 각각 Bax에 의한 toxicity를 저해하는 유전자를 찾기 위해 스크린되었다. 이러한 방법으로 정소 cDNA library 탐색에서는 5 $\times$ $10^{6}$ transformant중 39개, 난소cDNA library 탐색에서는 2 $\times$ $10^{6}$ transformant중 26개의 콜로니가 생존하였다. 이들 콜로니로부터 유전자를 분리하여 분석해 본 결과 여러 그룹으로 분류할 수 있었다. 각 그룹의 관련 유전자는 protein synthesis/degradation 12종, oxidation/reductation 5종, detoxin/ cell cycle promoter 3종, signal transduction/growth factor 5종, 그리고 알려지지 않은 유전자 9종이었다. 그 중, bax-toxicity inhibition에 강력한 survival phenotype을 가지는 유전자(pSEDL)를 동정하였다. 이것은 T3-4-1 콜로니로부터 분리하였는데 140개 아미노산으로 이루어진 인간 SEDL(GenBank, XM_013096) 유전자와 매우 유사한 homology를 가지며, bax와 관련된 기능은 밝혀져 있지 않다. 이외에도 분리된 유전자에는 NADH, thioreduction, 그리고 cytochrome oxidase와 같은 positive 유전자 군이 크로닝되어, Bax를 이용한 효모에서 apoptosis suppressor에 관련된 유전자를 손쉽게 스크린하는 것이 가능하고, 분리된 유전자의 기능을 예측할 수 있어 지금까지 보고된 유전자 크로닝법 보다는 강력한 수단으로 활용될 수 있다는 사실을 시사하였다. 그러나, ORF에 관계없이 Bax 발현에 저항하는 유전자군이 선발된다든지 하는 문제점은 금후 검토가 필요하리라 사료된다.

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Metabolic Activation of Marijuana Constituents, Cannabinoids, in Relation to Their Toxicity for Human and Its Oxidation Mechanism

  • Ikuo, Yamamoto
    • Proceedings of the PSK Conference
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    • 2002.10a
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    • pp.194-199
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    • 2002
  • Many oxidative metabolites of tetrahydrocannabinols (THCs), active components of marijuana, were pharmacologically active, and 11-hydroxy-THCs, 11-oxo-${\Delta}^8$-THC, 7-oxo-${\Delta}^8$-THC, 8$\beta$, 9$\beta$-epoxyhexahydrocannabinol (EHHC), 9$\alpha$, l0$\alpha$-EHHC and 3'-hydroxy-${\Delta}^9$-THC were more active than THC in pharmacological effects such as catalepsy, hypothermia and barbiturate synergism in mice. Cannabidiol (CBD), another major component, was biotransfomred to two novel metabolites, 6-hydroxymethyl-${\Delta}^9$-THC and 3-pentyl-6, 7, 7a, 8, 9, lla-hexahydro-I, 7-dihydroxy-7, 1O-dimethyldibenzo[b, d]oxepin (PHDO) through 8R, 9-epoxy-CBD and 85, 9-epoxy-CBD, respectively. Both metabolites exhibited some pharmacological effects comparable to d9 - THe. Cannabinol (CBN), the other major component, was mainly metabolized to ll-hydroxy-CBN by hepatic microsomes of animals including humans. The pharmacological effects of the metabolite were higher than those of CBN demonstrating that II-hydroxylation of CBN is metabolic activation pathway of the cannabinoid as is the case in THCs. Tolerance and reciprocal cross-tolerance developed to pharmacological effects d8 - THC and ll-hydroxy-d8-THC , and the magnitude of tolerance development produced by the metabolite was significantly higher than that by d8-THC. The results indicate that ll-hydroxy-d8-THC has an important role not only in the pharmacological effects but also its tolerance development of d8 - THe. THCs and their metabolites competed to the specific binding of CP-55, 940, an agonist of cannabinoid receptor, to synaptic membrane from bovine cerebral cortex. The Ki value of THCs and their metabolites were closely paralleled to their pharmacological effects in mice. A novel cytochrome P450 (cyp2c29) was purified and identified as a major enzyme responsible for the metabolic activation of d8-THC at the II-position in the mouse liver. cDNA of CYP2C29 was cloned from a mouse cDNA library and its sequence was determined. The oxidation mechanism of THC by cyp2c29 was proposed.

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Preliminary Application of Molecular Monitoring of the Pacific Herring (Clupea pallasii) Based on Real-time PCR Assay Utilization on Environmental Water Samples

  • Kim, Keun-Yong;Heo, Jung Soo;Moon, Seong Yong;Kim, Keun-Sik;Choi, Jung-Hwa;Yoo, Joon-Taek
    • Korean Journal of Ecology and Environment
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    • v.54 no.3
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    • pp.209-220
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    • 2021
  • Pacific herring, Clupea pallasii, a keystone species with significant ecological and commercial importance, is declining globally throughout much of its range. While traditional fishing equipment methods remain limited, new sensitive and rapid detection methods should be developed to monitor fisheries resources. To monitor the presence and quantity of C. pallasii from environmental DNA (eDNA) extracted from seawater samples, a pair of primers and a TaqMan® probe specific to this fish based on mitochondrial cytochrome b (COB) sequences were designed for the real-time PCR (qPCR) assay. The combination of our molecular markers showed high specificity in the qPCR assay, which affirmed the success of presenting a positive signal only in the C. pallasii specimens. The markers also showed a high sensitivity for detecting C. pallasii genomic DNA in the range of 1 pg~100 ng rxn-1 and its DNA plasmid containing COB amplicon in the range of 1~100,000copies rxn-1, which produced linear standard calibration curves (r2=0.99). We performed a qPCR assay for environmental water samples obtained from 29 sampling stations in the southeastern coastal regions of South Korea using molecular markers. The assay successfully detected the C. pallasii eDNA from 14 stations (48.2%), with the highest mean concentration in Jinhae Bay with a value of 76.09±18.39 pg L-1 (246.20±58.58 copies L-1). Our preliminary application of molecular monitoring of C. pallasii will provide essential information for efficient ecological control and management of this valuable fisheries resource.