• 제목/요약/키워드: Metabolizing Enzyme

검색결과 223건 처리시간 0.023초

ZR-75-1 human breast cancer cells th study the mechanism of action of PAHs

  • Nano, Min-Kyung;Yhong, Sheen-Yhun
    • 한국환경독성학회:학술대회논문집
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    • 한국환경독성학회 2002년도 추계국제학술대회
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    • pp.171-171
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    • 2002
  • Recent industrial society has human widely exposed to PAHs that are comming from the incomplete combustion of organic material as widerspread environmetal contaminants. Biological activities of PAHs are not known although PAHs are considered as carcinogens. PAHs in the mammalian cells affect CYP1A1 gene expression as well as other phase II drug metabolizing enzymes as UDPGT, NMOR etc. The mechanism of action of PAHs has been studied extensively, however it is not clear how PAHs turn on CYP1A1 in human breast cancer. Our labolatory have been studied the effect of PAHs in the human breast cancer cell lind MCF7. In this study, we examined the ZR-75-1 human breast cancer cells as a new system to evaluate bioactivity of PAHs. ZR-75-1 human breast cancer cell line has been estabilished from the breast cnacer patient, has estrogen receptors and progesteron receptors. We have been able to estbilish long term culture system of this cells then used for the study to observe the effect of PAHs. We demonstrate that PAHs induced the transcription of an aryl hydrocarbon-responsive reporter vector containing the CYP1A1 promoter and 7-ethoxyresolufin O-deethylase(EROD) activity of CYP1A1 enzyme in a concentration-dependant manner. RT-PCR analysises indicated that PAHs significantly up-regulate the constitutive level of CYP1A1 mRNA. Apparently, ZR-75-1 cells have Aryl hydrocarbon recetors, therefore it would be good experimental tool to study the cross-talk between PAHs and steroid actions.

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Isolation and Molecular Analysis of Methanol Oxidation Genes in an Obligate Methylotrophic Bacterium, Metheylobacillus sp. Strain SK-5

  • Choi, Hack-Sun;Kim, Jin-Kwon;Ahn, Yeong-Hee;Koh, Moon-Joo;Kim, Si-Wouk
    • Journal of Microbiology and Biotechnology
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    • 제12권5호
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    • pp.819-825
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    • 2002
  • Methanol dehydrogenase (MDH) is a key enzyme in the process of methanol oxidation in methylotrophic bacteria. However, information on MDH genes from genus Methylobacillus is limited. In this study, a 6.5-kb HindIII DNA fragment of Methylobacillus sp. SK-5 chromosomal DNA was isolated from the genomic library of the strain by using a degenerate oligonucleotide probe that was designed based on JV-terminal amino acid sequence of the MDH $\alpha$ subunit purified from the strain. Molecular analysis of the fragment revealed four tightly clustered genes (mxaFJGI) involved in the methanol oxidation. The first and fourth genes were very similar to mxaF (77% identity for nucleotides an 78% identity for amino acids) and mxaF (67% Identity for nucleotides and 68% Identity for amino acids) genes, respectively, from Methylovorus sp. SSI. Genes mxaF and mxaI encode $\alpha$ and $\beta$ subunits of MDH, respectively. The two subunits were identified from purified MDH from Methylobacillus sp. SK-5. A dendrogram constructed by comparison of amino acid sequences of MDH u subunits suggests that MxaF from Methylobacillus sp. SK-5 belongs to a subfamily cluster of MDH u subunits from $\beta$-subgroup Proteobacteria. The subfamily cluster is separated from the other subfamily that consists of $\beta$- and $\gamma$-subgroup Proteobacteria. This study provided information on mn genes from a methylotrophic bacterium in $\beta$-subgroup Proteobacteria, which would aid to better develop a gene probe to detect one-carbon metabolizing bacteria.

Alteration of Substrate Specificity by Common Variants, E158K/E308G and V257M, in Human Hepatic Drug-metabolizing Enzyme, Flavin-containing Monooxygenase 3

  • Lee, Jung-Kyu;Kang, Ju-Hee;Cha, Young-Nam;Chung, Woon-Gye;Park, Chang-Shin
    • The Korean Journal of Physiology and Pharmacology
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    • 제7권3호
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    • pp.157-162
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    • 2003
  • Our earlier studies found a significant correlation between the activities of ranitidine N-oxidation catalyzed by hepatic flavin-containing monooxygenase (FMO) and the presence of mutations in exon 4 (E158K) and exon 7 (E308G) of the FMO3 gene in Korean volunteers. However, caffeine N-1 demethylation (which is also partially catalyzed by FMO) was not significantly correlated with these FMO3 mutations. In this study, we examined another common mutation (V257M) in exon 6 of FMO3 gene. The V257M variant, which is caused by a point mutation (G769A), was commonly observed (13.21% allele frequency) in our subjects (n=159). This point mutation causes a substitution of $Val^{257}$ to $Met^{257}$, with transformation of the secondary structure. The presence of this mutant allele correlated significantly with a reduction in caffeine N-1-demethylating activity, but was not correlated with the activity of N-oxidation of ranitidine. In a family study, the low FMO activity observed in a person heterozygous for a nonsense mutation in exon 4 (G148X) and heterozygous for missense mutation in exon 6 (V257M) of FMO3 was attributed to the mutations. Our results suggest that various point mutations in the coding regions of FMO3 may influence FMO3 activity according to the probe substrates of varying chemical structure that correlate with each mutation on the FMO3 gene.

Inhibition of Carcinogen-Activating Cytochrome P450 Enzymes by Xenobiotic Chemicals in Relation to Antimutagenicity and Anticarcinogenicity

  • Shimada, Tsutomu
    • Toxicological Research
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    • 제33권2호
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    • pp.79-96
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    • 2017
  • A variety of xenobiotic chemicals, such as polycyclic aromatic hydrocarbons (PAHs), aryl- and heterocyclic amines and tobacco related nitrosamines, are ubiquitous environmental carcinogens and are required to be activated to chemically reactive metabolites by xenobiotic-metabolizing enzymes, including cytochrome P450 (P450 or CYP), in order to initiate cell transformation. Of various human P450 enzymes determined to date, CYP1A1, 1A2, 1B1, 2A13, 2A6, 2E1, and 3A4 are reported to play critical roles in the bioactivation of these carcinogenic chemicals. In vivo studies have shown that disruption of Cyp1b1 and Cyp2a5 genes in mice resulted in suppression of tumor formation caused by 7,12-dimethylbenz[a]anthracene and 4-(methylnitrosamino)-1-(3-pyridyl)-1-butanone, respectively. In addition, specific inhibitors for CYP1 and 2A enzymes are able to suppress tumor formation caused by several carcinogens in experimental animals in vivo, when these inhibitors are applied before or just after the administration of carcinogens. In this review, we describe recent progress, including our own studies done during past decade, on the nature of inhibitors of human CYP1 and CYP2A enzymes that have been shown to activate carcinogenic PAHs and tobacco-related nitrosamines, respectively, in humans. The inhibitors considered here include a variety of carcinogenic and/or non-carcinogenic PAHs and acethylenic PAHs, many flavonoid derivatives, derivatives of naphthalene, phenanthrene, biphenyl, and pyrene and chemopreventive organoselenium compounds, such as benzyl selenocyanate and benzyl selenocyanate; o-XSC, 1,2-, 1,3-, and 1,4-phenylenebis(methylene)selenocyanate.

약물대사효소 CYP2C19, CYP2D6의 다형성과 사상체질의 관련성 연구 (Association between Genetic Polymorphisms of the CYP2C19, CYP2D6 and Types of Sasang Constitutional Medicine)

  • 이상규;김현주;박혜정;이정호;권덕윤;주종천;최선미;이혜숙;김윤경
    • 대한한의학회지
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    • 제28권1호통권69호
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    • pp.51-62
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    • 2007
  • Objectives . The types of Sasang constitutional medicine (SCM) have definite effect on response to herbal drugs. The majority of human P45O dependent xenobiotic metabolism is carried out by polymorphic enzymes which can cause abolished, altered or enhanced metabolism. Therefore, we evaluated the relation of major CYP2C19, 2D6 polymorphism with Sasang types. Methods : 214 healthy subjects were recruited with informed consent; 172 among them had Sasang diagnosis by QSCC2. CYP2D6, 2C19 polymorphism were determined by PCR-RFLP method. Results : None of the Sasang types showed significant difference in CYP2D6, 2C19 polymorphism. However, the Tae-um type showed relatively low frequency of CYP2D6 $^{*}$10/$^{*}$10 polymorphisms with low activity (p=0.110). In the So-yang type, specific $^{*}$3/$^{*}$3 genotype which is a poor metabolizer of CYP2C19$^{*}$3 was detected (p=0.078).Conclusion . These results suggest that the Tae-um type which is said to have high liver function in SCM has the tendency of high drug-metabolizing enzyme activity. With further study, the CYP polymorphism could serve as a scientific tool for SCM diagnosis.

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Effects of Methyl Gallate on Arachidonic Acid Metabolizing Enzymes: Cyclooxygenase-2 and 5-Lipoxygenase in Mouse Bone Marrow-Derived Mast Cells

  • Kim, Se-Jong;Jin, Mei-Hua;Lee, Eun-Kyung;Moon, Tae-Chul;Quan, Zhe-Jiu;Yang, Ju-Hye;Son, Kun-Ho;Kim, Kil-Ung;Son, Jong-Kun;Chang, Hyeun-Wook
    • Archives of Pharmacal Research
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    • 제29권10호
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    • pp.874-878
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    • 2006
  • Methyl gallate (MG) is a medicinal herbal product that is isolated from Paeonia lactiflora that inhibits cyclooxygenase-2 (COX-2) dependent phases of prostaglandin $D_2\;(PGD_2)$ generation in bone marrow-derived mast cells (BMMC) in a concentration-dependent manner with an $IC_{50}$ values of $17.0\;{\mu}M$. This compound also found inhibited the COX-2-dependent conversion of the exogenous arachidonic acid to $PGD_2$ in a dose-dependent manner with an $IC_{50}$ values of $190\;{\mu}M$, using a COX enzyme assay kit. However, at concentrations up to $80\;{\mu}M$, MG did not inhibit COX-2 protein expression in BMMC, indicating that MG inhibits COX-2 activity directly. Furthermore, MG consistently inhibited the production of leukotriene $C_4\;(LTC_4)$ in a dose dependent manner, with an $IC_{50}$ value of $5.3\;{\mu}M$. These results demonstrate that MG has a dual cyclooxygenase-2/5-lipoxygenase inhibitory activity, which might provide the basis for novel anti-inflammatory drugs.

Benzo(a)pyrene 유도 DNA 손상에 대한 Genistein과 청국장추출물의 보호효과 (Protective Effect of Genistein and Korean Fermented Soybean (Chungkookjang) Extract against Benzo(a)pyrene Induced DNA Damage in HepG2 Cells)

  • 송은정;김현표;허문영
    • 약학회지
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    • 제52권5호
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    • pp.376-383
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    • 2008
  • Chungkookjang (CKJ) is a fermented soybean product and one of favorite traditional foods in Korea. In this study, the alcoholic extract from Korean fermented soybean (CKJ) and its one of major flavonoids, genistein were evaluated for their protective effect against B(a)P induced cytotoxicity and DNA damage in HepG2 cells. CKJ extract and genistein decreased B(a)P-induced cell cytotoxicity. CKJ extract inhibited DNA single strand breaks evaluated by single cell gel electrophoresis. From RT-PCR study, it was revealed that CKJ extract decrease DNA damage induced in HepG2 cells expressing CYP1A1 and 1A2 by B(a)P. The metabolizing activities of CYP1A1 and CYP1A2, as measured by the 7-alkoxy resorufin O-deethylation (AROD) assay, showed that CKJ extract and genistein inhibited CYP1A1 and CYP1A2 activities. Genistein may contribute to these biological effects of CKJ extract at least in part. All these results indicate that CKJ extract and genistein may be useful for protection against B(a)P-induced cytotoxicity and DNA damage. Therefore, the alcoholic extract of Korean fermented soybean (CKJ) is suggested to be promising functional food which can prevent the cellular genotoxicity of dietary and lifestyle related carcinogens.

Interaction of the Lysophospholipase PNPLA7 with Lipid Droplets through the Catalytic Region

  • Chang, Pingan;Sun, Tengteng;Heier, Christoph;Gao, Hao;Xu, Hongmei;Huang, Feifei
    • Molecules and Cells
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    • 제43권3호
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    • pp.286-297
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    • 2020
  • Mammalian patatin-like phospholipase domain containing proteins (PNPLAs) play critical roles in triglyceride hydrolysis, phospholipids metabolism, and lipid droplet (LD) homeostasis. PNPLA7 is a lysophosphatidylcholine hydrolase anchored on the endoplasmic reticulum which associates with LDs through its catalytic region (PNPLA7-C) in response to increased cyclic nucleotide levels. However, the interaction of PNPLA7 with LDs through its catalytic region is unknown. Herein, we demonstrate that PNPLA7-C localizes to the mature LDs ex vivo and also colocalizes with pre-existing LDs. Localization of PNPLA7-C with LDs induces LDs clustering via non-enzymatic intermolecular associations, while PNPLA7 alone does not induce LD clustering. Residues 742-1016 contains four putative transmembrane domains which act as a LD targeting motif and are required for the localization of PNPLA7-C to LDs. Furthermore, the N-terminal flanking region of the LD targeting motif, residues 681-741, contributes to the LD targeting, whereas the C-terminal flanking region (1169-1326) has an anti-LD targeting effect. Interestingly, the LD targeting motif does not exhibit lysophosphatidylcholine hydrolase activity even though it associates with LDs phospholipid membranes. These findings characterize the specific functional domains of PNPLA7 mediating subcellular positioning and interactions with LDs, as wells as providing critical insights into the structure of this evolutionarily conserved phospholipid-metabolizing enzyme family.

유기게르마늄(GE-132)이 Bromobenzene의 대사계에 미치는 영향 (Effect of GE-132 on the Hepatic Bromobenzene Metabolizing Enzyme System in Rats)

  • 김석환;조태현;최종원
    • 한국식품영양과학회지
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    • 제22권6호
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    • pp.702-708
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    • 1993
  • Bromobenzene투여시 GE-132의 해독기전을 추구할 목적으로 epoxide생성계와 해독계 효소 및 glutathione 관련 효소계의 활성에 GE-132가 어떤 영향을 주는가를 검토하여 다음과 같은 실험 결과를 얻었다. GE-132(100mg/kg)의 전처리로 cytochrome P-450, amino=pyrine demethylase 및 aniline hydroxylase와 해독계 효소인 epoxide hydrolase활성에는 영향을 미치지 않았으나, glutathions S-transferase활성은 증가 하였다. Bromobenzene(460mg/kg)을 주사하였을때 glutathione S-transferase의 활성이 현저히 감소되던 것이 GE-132의 투여로 대조군 수준으로 증가되었다. 조직내 glutathione의 함량변동도 bromobenzene 투여로 감소되던 것이 GE-132의 전처리로 bromobenzene단독 투여군보다 증가되었으며 ${\gamma}-glutamylcystein$ synthtase의 활성은 각 실험군에서 별다른 영향이 없는데 비해 glutathione reductase의 활성은 bromobenzene투여로 억제 되던 것이 GE-132의 전처리로 대조군 수준으로 활성을 유지하고 있었다.

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국산 양식 홍합 함유 식빵의 제조 및 생리활성 평가 (Preparation and Evaluation of Physiological Activity of White Bread Containing Korean Blue Mussel)

  • 조민지;김윤아;이승철
    • 한국식품영양과학회지
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    • 제45권11호
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    • pp.1623-1629
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    • 2016
  • 우리나라 남해안에서 많이 양식되고 있는 홍합(진주담치)은 다가 고도 불포화지방산을 비롯한 다양한 유용 물질을 함유하고 있으며 이로 인한 항산화능, 항비만능, 알코올분해 효소 촉진능 등의 생리활성이 보고되었다. 본 연구에서는 홍합의 이용성 다양화를 위하여 홍합 분말을 첨가한 식빵(0, 0.25, 0.5, 0.75, 1%, 홍합/밀가루, w/w)을 직접반죽법으로 제조하여 그 특성을 분석하였다. 홍합분말이 첨가될수록 식빵의 명도는 감소하였고, 적색도와 황색도는 증가하였다. 홍합분말의 첨가는 식빵의 비용적에 크게 영향을 주지 않았지만, 항산화능(DPPH 및 ABTS 라디칼 소거능)은 증가시켰다. 또한, 홍합분말의 함량이 증가할수록 알코올 탈수소효소와 아세트알데히드 탈수소효소의 활성도 증가하였다. 관능 검사에서는 홍합분말 1% 첨가 식빵이 맛, 향, 물성과 선호도에서 상대적으로 높은 점수를 얻었다. 이상의 결과는 홍합이 식빵의 경도와 강도에 큰 영향을 주지 않고 생리활성을 향상 할 수 있는 소재로 활용될 수 있음을 의미한다.