• Title/Summary/Keyword: P450nor

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Enzyme Activity and Gene Expression of Cytochrome P450 Involved in Capsidiol Biosynthesis in Solanaceae Plants (가지과식물에서 Capsidiol 생합성에 관여하는 Cytochrome P450 유전자의 발현과 효소활성)

  • Kwon, Soon-Tae;Hasegawa, Paul
    • Korean Journal of Plant Resources
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    • v.21 no.2
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    • pp.139-143
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    • 2008
  • Enzyme activity and expression of cytochrome P450 gene involved in the pathway of capsidiol biosynthesis were compared in five different solanaceae plants such as red pepper, green pepper, tobacco, potato and egg plant. Base on genomic DNA and/or RT-PCR results, four solanaceae plants such as red pepper, green pepper, tobacco and egg plant possess P450 gene in the genome and specifically expressed by elicitor treatment. However, potato was appeared to have neither P450 nor cyclase gene in the genome. P450 genes did not show any expression in the plants under normal condition, but showed highly specific expression under elicitation condition in various organs and tissue such as leaf, root, stem and culture cells.

Cytochrome P450 monooxygenase analysis in free-living and symbiotic microalgae Coccomyxa sp. C-169 and Chlorella sp. NC64A

  • Mthakathi, Ntsane Trevor;Kgosiemang, Ipeleng Kopano Rosinah;Chen, Wanping;Mohlatsane, Molikeng Eric;Mojahi, Thebeyapelo Jacob;Yu, Jae-Hyuk;Mashele, Samson Sitheni;Syed, Khajamohiddin
    • ALGAE
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    • v.30 no.3
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    • pp.233-239
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    • 2015
  • Microalgae research is gaining momentum because of their potential biotechnological applications, including the generation of biofuels. Genome sequencing analysis of two model microalgal species, polar free-living Coccomyxa sp. C-169 and symbiotic Chlorella sp. NC64A, revealed insights into the factors responsible for their lifestyle and unravelled biotechnologically valuable proteins. However, genome sequence analysis under-explored cytochrome P450 monooxygenases (P450s), heme-thiolate proteins ubiquitously present in species belonging to different biological kingdoms. In this study we performed genome data-mining, annotation and comparative analysis of P450s in these two model algal species. Sixty-nine P450s were found in two algal species. Coccomyxa sp. showed 40 P450s and Chlorella sp. showed 29 P450s in their genome. Sixty-eight P450s (>100 amino acid in length) were grouped into 32 P450 families and 46 P450 subfamilies. Among the P450 families, 27 P450 families were novel and not found in other biological kingdoms. The new P450 families are CYP745-CYP747, CYP845-CYP863, and CYP904-CYP908. Five P450 families, CYP51, CYP97, CYP710, CYP745, and CYP746, were commonly found between two algal species and 16 and 11 P450 families were unique to Coccomyxa sp. and Chlorella sp. Synteny analysis and gene-structure analysis revealed P450 duplications in both species. Functional analysis based on homolog P450s suggested that CYP51 and CYP710 family members are involved in membrane ergosterol biosynthesis. CYP55 and CYP97 family members are involved in nitric oxide reduction and biosynthesis of carotenoids. This is the first report on comparative analysis of P450s in the microalgal species Coccomyxa sp. C-169 and Chlorella sp. NC64A.

Effects of the Genetic Polymorphisms on Urinary Excretion of 1-Hydroxypyrene and 2-Naphthol (일반인구에서 유전자 다형성이 요중 1-hydroxypyrene 및 2-naphthol의 배설량에 미치는 영향)

  • Hwang Moon-Young;Cho Byung-Mann;Moon Seong-Bae
    • Journal of Environmental Science International
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    • v.14 no.5
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    • pp.499-511
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    • 2005
  • This study was performed to determine the effects of genetic polymorphisms, such as glutathione S-transferase ${\mu}1(GSTM1)$, glutathione S-transferase ${\Theta}1\;(GSTM1)$, glutathione S-transferase ${\pi}l (GSTP1)$, aryl hydrocarbon N-acetyltransferase 2 (NAT2), cytochrome P450 2E1 (CYP2E1), cytochrome P450 1A1 (CYP1A1) on the concentrations of urinary 1-hydroxypyrene (1-OHP) and 2-naphthol in general population with no occupational exposure to polycyclic aromatic hydrocarbons (PAHs). Study subjects were 257 men who visited a health promotion center in Susan. A questionnaire was used to obtain detailed data about age, smoking, drinking, body fat mass, intake of fat etc. Urinary l-OHP and 2-naphthol concentration were analyzed by HPLC system with a fluorescence detector. A multiplex PCR method was used to identify the genotypes for GSTM1 and GSTT1. The polymorphisms of GSTP1, NAT2, CYP1A1 and CYP2E1 were determined by the polymerase chain reaction restriction fragment length polymorphism (PCR-RFLP) method. Urinary 1-OHP concentration was higher in deleted genotype of GSTM1, increased as smoking and alcohol drinking increased. Urinary 2-naphthol concentration was also rely on the age and smoking. Neither genetic polymorphism nor drinking-related factors were significantly related to urinary 2-naphthol concentration. No significant relation was found between physical characteristics and concentrations of urinary PAHs metabolites in the subjects, but the geometric mean of urinary 1-OHP and 2-naphthol was higher in the group with higher value compared to median value. These data suggest that in general population occupationally not exposed to PAHs, urinary concentration of PAHs metabolites is influenced by smoking, alcohol drinking and deleted genotype of GSTM1 in 1-OHP and smoking in 2-naphthol.

Differential Growth Response and Gene Expression in Relation to Capsidiol Biosynthesis of Red Pepper Plant and Cultured Cells by γ-Ray and UV Stress (방사선과 자외선에 대한 고추 식물체 및 배양세포의 생장반응과 Capsidiol 생합성 유전자의 발현 차이)

  • An, Jung-Hee;Kim, Jae-Sung;Jeong, Jeong-Hag;Oh, Sei-Myoung;Kwon, Soon-Tae
    • Journal of Plant Biotechnology
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    • v.30 no.2
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    • pp.201-206
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    • 2003
  • Differential responses of red pepper plant and cultured cells to enhanced ${\gamma}$-ray($^{60}$ Co) and ultraviolet(UV) stress were investigated. In seed treatment, 1 Gy of ${\gamma}$-ray increased seedling dry weight up to 19.1%, but 50 Gy treatment markedly ingibited seed germination and subsequent growth of seedling. UV treatment to seed did not change the germination ability of seeds and the growth of seedlings regardless of duration of UV treatment until 24 hrs. In case of UV treatment to seedlings, plant injury was seriously progressed even after the seedlings were returned to no UV condition, and eventually all the leaves showed chlorosis by the stress. However, progress of plant injury by ${\gamma}$-ray stress slower than that caused by UV stress, and even at the high dose of ${\gamma}$-ray 50 Gy, did not caused the cholrosis of stressed plant leaf. Amount of electrolytes leakage from plant leaf by UV treatment for 24hrs was increased up to 28.8 folds in comparison with untreated control, whereas that of 50 Gy of ${\gamma}$-ray was increased only 1.2 folds. UV stress induced the production of capsidiol, antimicrobial phytoalexin, by activation of gene expression involved in capsidiol biosynthesis, such as sesquiterpene cyclase and cyclase and cytochrome P450 hydroxylase in the leaf and cultured cell, but ${\gamma}$-ray stress induced neither the production of capsidiol nor expression of the genes.

Protocols for Estrus Synchronization in Awassi Ewes under Arid Environmental Conditions

  • Abdullah, A.Y.;Husein, M.Q.;Kridli, R.T.
    • Asian-Australasian Journal of Animal Sciences
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    • v.15 no.7
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    • pp.957-962
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    • 2002
  • In July, three trials were conducted to evaluate the best sponge type and optimum PMSG dose to be administered to sheep under the Jordanian Badia (arid) conditions. In trial 1, three flocks (n=77, n=18 and n=47 for flocks 1, 2 and 3, respectively) were administered with 40 mg fluorogestone acetate (FGA) intravaginal sponges for 12 days before receiving 600 IU of PMSG at the time of sponge removal. In trial 2, 95 ewes were assigned to 4 groups to receive 300 (n=25), 450 (n=27), 600 (n=22) or 750 (n=21) IU of PMSG following a 12 d FGA 40 mg sponge insertion period. In trial 3, 60 ewes were assigned to 3 groups (n=20) to receive either FGA 30 mg, FGA 40 mg or medroxyprogesterone acetate (MAP) 60 mg intravaginal sponges all followed by an administration of 600 IU of PMSG at sponge removal. In all trials, rams were isolated 1 day before sponge insertion and were allowed back with the ewes at sponge removal. Estrual responses and lambing data were collected. The effects of treatment, milking status and face color on estrual responses and lambing data were examined. In trial 1, greater first cycle conception rate (p<0.05), twinning rate (p<0.01) and the number of lambs born/served ewe (p<0.01) were observed in flock 2 compared with flocks 1 and 3. Neither face color nor milking status had any influence on the measured parameters (p>0.05). Despite low lambing rate in trial 2, ewes receiving 600 IU of PMSG had greater (p<0.05) number of lambs born/served ewe compared with ewes receiving 450 IU of PMSG. Regardless of PMSG dose, intervals to detected estrus occurred 10 h earlier (p<0.01) in dry than lactating ewes. Similar to trial 2, lambing rate was depressed in trial 3. The expression of estrus was advanced (p<0.05) in ewes receiving MAP 60 mg sponges compared with those receiving FGA 30 and FGA 40 mg sponges (42$\pm$3.1, 49$\pm$3.1 and 49$\pm$3.1 h post sponge removal in ewes receiving MAP 60 mg, FGA 30 mg and FGA 40 mg sponges, respectively). Other parameters were not influenced (p>0.05) by sponge type, milking status and face color. Data show that a 600 IU dose of PMSG tends to give the best lambing results. In addition, results indicate that the use 60 mg MAP sponges for estrus synchronization may be more appropriate under the Jordanian Badia conditions during late seasonal anestrus.

CYP2E1 rs2031920, COMT rs4680 Polymorphisms, Cigarette Smoking, Alcohol Use and Lung Cancer Risk in a Japanese Population

  • Kakino, Kenichi;Kiyohara, Chikako;Horiuchi, Takahiko;Nakanishi, Yoichi
    • Asian Pacific Journal of Cancer Prevention
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    • v.17 no.8
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    • pp.4063-4070
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    • 2016
  • Background: Cytochrome P450 2E1 (CYP2E1) and catechol-O-methyltransferase (COMT) genes may contribute to susceptibility to lung cancer because of their critical involvement in mechanisms of carcinogenesis. Materials and Methods: We evaluated the role of CYP2E1 rs2031920 and COMT rs4680 in a case-control study involving 462 lung cancer cases and 379 controls in Japanese. Logistic regression was used to assess adjusted odds ratios (OR) and 95% confidence intervals (CI). Multiplicative and additive interactions with cigarette smoking or alcohol use were also examined. Results: Neither CYP2E1 rs2031920 nor COMT rs4680 was associated with lung cancer risk overall. However, smokers with the CC genotype of CYP2E1 rs2031920 (OR = 3.57, 95% CI = 2.26 - 5.63) presented a higher risk of lung cancer than those with at least one T allele (OR = 2.91, 95% CI = 1.70 - 4.98) as compared to never-smokers with at least one T allele (reference). Subjects with excessive drinking and the CC genotype of CYP2E1 rs2031920 had a significantly higher risk (OR = 2.22, 95% CI =1.39 - 3.56) than appropriate drinkers with at least one T allele. A similar tendency was observed between COMT rs4680 and either smoking or drinking habits. There were no multiplicative or additive interactions between the polymorphisms and either smoking or alcohol use. Conclusions: Our findings indicate that CYP2E1 rs2031920 and COMT rs4680 are not major contributors to lung cancer risk in our Japanese population. Future studies on the genetics of lung cancer in Japanese and their environment interactions are required.

Effects of the Peroxisome Proliferator Ciprofibrate and Prostaglandin $F_2\alpha$ Combination Treatment on Second Messengers in Cultured Rat Hepatocytes

  • Hong, Jin-Tae;Yun, Yeo-Pyo
    • Archives of Pharmacal Research
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    • v.21 no.2
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    • pp.120-127
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    • 1998
  • Peroxisome proliferators induce hepatic peroxisome proliferation and hepatic tumors in rodents. These chemicals increase the expression of the peroxisomal $\beta$-oxidation pathway and the cytochrome P-450 4A family, which metabolizes lipids, including eicosanoids. Peroxisome proliferators transiently induce increased cell proliferation in vivo. However, peroxisome proliferators are weakly mitogenic and are not co-mitogenic with epidermal growth factor (EGF) in cultured hepatocytes. Earlier study found that the peroxisome proliferator ciprofibrate is cornitogenic with eicosanoids. In order to study possible mechanisms of the comitogenicity of peroxisome proliferator ciprofibrate and eicosanoids' we hypothesized that the co-mitogenicity may result from synergistic or additive increases of second messengers in mitogenic signal pathways. We therefore examined the effect of the peroxisome proliferator ciprofibrate, prostaglandin $F_2_{\alpha}$($PGF_2{\alpha}$) and the combination of ciprofibrate and $PGF_2{\alpha}$ with or without growth factors on the protein kinase C (PKC) activity, and inositol-1, 4, 5-triphosphate ($IP_{3-}$) and intracellular calcium ($[Ca^{2+}]_i$) concentrations in cultured rat hepatocytes. The combination of ciprofibrate and $PGF_2{\alpha}$ significantly increased particulate PKC activity. The combination of ciprofibrate and $PGF_2{\alpha}$ also significantly increased EGF, transforming growth factor-$\alpha$ ($TGF_2{\alpha}$) and hepatic growth factor (HGF)-induced particulate PKC activity. The combination of ciprofibrate and $PGF_2_\alpha$greatly increased $[Ca^{2+}]_i$. However, the increases of PKC activity and $[Ca^{2+}]_i$ by ciprofibrate and $PGF_2{\alpha}$ alone were much smaller. Neither ciprofibrate or $PGF_2{\alpha}$ alone nor the combination of ciprofibrate and $PGF_2{\alpha}$ significantly increased the formation of $IP_3$. The combination of ciprofibrate and $PGF_2{\alpha}$, however, blocked the inhibitory effect of $TGF-{\beta}$ on particulate PKC activity and formation of $IP_3$ induced by EGF. These results show that co-mitogenicity of the peroxisome proliferator ciprofibrate and eicosanoids may result from the increase in particulate PKC activity and intracellular calcium concentration but not from the formation of $IP_3$.

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Mechanism of Relaxation of Rat Aorta by Scopoletin; an Active Constituent of Artemisia Capillaris

  • Kwon Eui Kwang;Jin Sang Sik;oChoi Min H;Hwang Kyung Taek;Shim Jin Chan;Hwang Il Taek;Han Jong Hyun
    • Journal of Physiology & Pathology in Korean Medicine
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    • v.16 no.2
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    • pp.389-396
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    • 2002
  • In the present work, we examined the mechanism of vasorelaxant effect of scopoletin, an active constituent of Artemisia capillaris on rat thoracic descending aortic rings. Scopoletin induced a concentration-dependent relaxation in rat thoracic descending aortic rings pre-contracted with phenylephrine (EC/sub 50/ = 238.94±37.4 μM), while it was less effective in rat thoracic descending aortic rings precontracted with high potassium solution (KCI 30 mM). Vasorelaxation by scopoletin was significantly inhibited after endothelial removal, but recovered at high concentration. Pretreatment of rat thoracic descending aortic rings with N/sup G/-nitro-L-arginine (100 μM), a nitric oxide synthase inhibitor, and atropine (1 μM), a muscarinic receptor antagonist, significantly inhibited scopoletin-induced relaxation of rat thoracic descending aortic rings. Neither indomethacin (3 μM), an inhibitor of cydooxygenase, nor propranolol (1 μM), a β -adrenoceptor antagonist, modified the effect of scopoletin. The combination of N/sup G/ -nitro-L-arginine (100 μ M) and miconazole (10 μ M), an inhibitor of cytochrome P 450, did not modify the effect of scopoletin, when compared with pretreatment with N/sup G/-nitro-L-arginine(100 μM) alone. Vasorelaxant effect of scopoletin was inverted by pretreatment with diltiazem (10 μM), a Ca/sup 2+/-channel blocker, at low concentration, while restored at high concentration. Apamin (K/sub ca/-channel blocker, 1 μM), 4-aminopyridine (4-AP, K/sub v/-channel blocker, 1 mM), and tetrodotoxin (TTX, Na/sup +/-channel blocker 1 μM) potentiated the vasorelaxant effect of scopoledn, but glibendamide (K/sub ATP/-channel blocker, 10 μM), tetraetylammonium(TEA, non-selective K-channel blocker, 10 mM) did not affect the relaxation of scopoletin. Free radical scavengers (TEMPO, catalase, mannitol) did not modify vascular tone. These results suggest that nitric oxide, Ca/sup 2+/ -channels play a role in endothelium-dependent relaxations to scopoletin in rat aortas, that apamin, 4-AP, TTX but not glibenclamide, TEA potentiated relaxation to scopoletin mediated by these channels, and that free radicals do not concern to the vasorelaxant effect of scopoletin.