• Biomolecules & Therapeutics
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• 제4권1호
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• pp.103-109
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• 1996

To investigate the effect of the third intracellular loop (i3 loop) peptide of human $\beta$$_2$-adrenergic receptor on receptor agonist binding, we expressed third intracellular loop region of human $\beta$$_2$-adrenergic receptor as glutathione S-transferase fusion protein in E. coli. DNA fragment of the receptor gene which encodes amino acid 221-274 of human $\beta$$_2$-adrenergic receptor was amplified by polymerase chain reaction and subcloned into the bacterial fusion protein expression vector pGEX-CS and expressed as a form of glutathione-S-transferase (GST) fusion protein in E. coli DH5$\alpha$. The receptor fusion protein was identified by SDS-PAGE and Western blot using monoclonal anti-GST antibody. The fusion protein expressed in this study was purified to an apparent homogeneity by glutathione Sepharose CL-4B affinity chromatography. The purified i3 loop fusion proteins at a concentration of 10 $\mu\textrm{g}$/ι caused right shift of the isoproterenol competition curve of [$^3$H]Dihydroalprenolol binding to hamster lung $\beta$$_2$-adrenergic receptor indicating lowered affinity of isoproterenol to $\beta$$_2$-adrenergic receptor possibly due to the uncoupling of receptor and G protein in the presence of the fusion protein. The uncoupling of receptor and G protein suggests that i3 loop region plays a critical role on $\beta$$_2$-adrenergic receptor G protein coupling.

• Preventive Nutrition and Food Science
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• 제10권4호
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• pp.340-343
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• 2005

Induction of NAD(P)H:(quinone-acceptor) oxidoreductase (QR) which promotes obligatory two electron reduction of quinones and prevents their participation in oxidative cycling and thereby the depletion of intracellular glutathione, has been used as a marker for chemopreventive agents. Induction of phase II enzyme is considered to be an important mechanism of cancer prevention. In our previous study, we assessed the quinone reductase QR-inducing activities of 216 kinds of medicinal herb extracts in cultured murine hepatoma cells, BPRc1 and hepalc1c7 cells. Among the 216 herbal extracts tested in that study, extracts from Chrysanthemum zawadskii showed significant induction of QR. In this study, we examined QR-inducing activity of solvent fractions of the herbal extract. The dichloromethane fraction of the herb showed the highest QR induction among the samples fractionated with four kinds of solvents with different polarity. The fraction also significantly induced the activity of glutathione S-transferase (GST), one of the major detoxifying enzymes, at $4{\mu}g/mL\;and\;2{\mu}g/mL$ in hepalc1c7 and BPRc1 cells, respectively. In conclusion, dichloromethane-soluble fraction of Chrysanthemum zawadskii which showed relatively strong induction of detoxifying enzymes merits further study to identify active components and evaluate their potential as cancer preventive agents.

• 한국균학회지
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• 제28권1호
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• pp.55-59
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• 2000

까치버섯(Polyozellus multiplex) 메탄올 추출물과 물 추출물은 산화 유발물질인 $H_2O_2$에 대하여 메탄올 추출물과 물 추출물에서 각각 53.5%, 29.4%의 생존율을 보여 까치버섯 추출물의 항산화 효과를 확인하였다. 까치버섯 메탄올 추출물과 물 추출물을 이용하여 동물에 강력한 발암물질인 MHNG를 투여한 후 항산화 효소계를 측정한 결과, 물 추출물은 항산화효소인 glutathione S-transferase(GST)와 superoxide dismutase(SOD) 활성과 조직내의 해독물질인 glutathione(GSH)의 함량을 높여 주었다. 또한 0.5%의 까치버섯 물 추출물은 암억제 유전자인 p53회 발현을 매우 증가시켰다. 이상의 결과로 까치버섯(Polyozellus multiplex)은 항산화 능이 있으며 특히, 물 추출물은 체내의 항산화 효소를 활성화시키고 항산화 물질의 함량을 높여주었고 암 억제유전자인 p53의 발현을 증가시켜 항암효과가 있음을 보여주었다.

• Nutrition Research and Practice
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• 제12권2호
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• pp.118-128
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• 2018

BACKGROUND/OBJECTIVES: Glutathione s-transferase (GST) is involved in the formation of a multigene family comprising phase II detoxification enzymes, involved in the detoxification of reactive oxygen species. This study evaluated whether daily supplementation with kale juice could modulate levels of plasma antioxidant vitamins and oxidative stress-related parameters. We further examined whether this modulation was affected by combined GSTM1 and T1 polymorphisms. SUBJECTS/METHODS: Totally, 84 subclinical hypertensive patients having systolic blood pressure (BP) over 130 mmHg or diastolic BP over 85 mmHg, received 300 mL of kale juice daily for 6 weeks. Blood samples were drawn before start of study and after completion of 6 weeks. RESULTS: After supplementation, we observed significant decrease in DNA damage and increase in erythrocyte catalase activity in all genotypes. Plasma level of vitamin C was significantly increased in the wild/null and double null genotypes. The plasma levels of ${\beta}-carotene$, erythrocyte glutathione peroxidase activity, and nitric oxide were increased only in the wild/null genotype after kale juice supplementation. CONCLUSIONS: The effect of kale juice was significantly greater in the GSTM1 null genotype and wild/null genotype groups, suggesting possibility of personalized nutritional prescriptions based on personal genetics.

• 한국약용작물학회지
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• 제15권1호
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• pp.21-25
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• 2007

해당화 뿌리는 우리나라 민간에서 당뇨병 치료제로 사용되는 약용식물이다. Bromobenzene으로 간독성을 유발한 흰쥐에 뿌리에서 분리한 화합물인 (+)-catechin을 경구투여하여 bromobenzene대사계에 미치는 효소활성을 간독성 물질인 bromobenzene 3,4-oxide 생성에 관여하는 효소인 aminopyrine N-demetylase와 aniline hydroxylase와 독성 epoxide 대사중간체를 무독화 시키는 epoxide hydrolase와 glutathione S-transferase에 활성을 관찰하였다. (+)-Catechin의 투여가 aminopyrine N-demetylase, aniline hydroxylase 및 glutathione S-transferase에 활성에는 영향을 주지 못하였으나, epoxide hydrolase는 positive control로 사용한 ascorbic acid에 미치지 못하지만, bromobenzene 처리군 보다 39% 효소활성을 회복 시켰다. 따라서, (+)-catechin은 간독성 물질을 무독화시키는 epoxide hydrolase의 활성을 회복시켜 간보호 활성을 나타냄을 알 수 있었으며, 해당화에서 분리한 사포닌 성분인 rosamultin도 이효소의 활성을 증가시킴으로 인해 보호활성을 나타내는 것으로 보고된바 있다.

• 생명과학회지
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• 제21권8호
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• pp.1076-1082
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• 2011

미세소관(microtubule) 위를 이동하는 키네신은 분비소포를 이동시키는 운동단백질이다. KIF5s (KIF5A, KIF5B and KIF5C)는 세포막으로 싸인 각종 세포 내 소기관과 결합하여 미세소관을 따라 목적지까지 이동시킨다는 결과는 알려져 있지만, 어떻게 상대의 cargo를 인식하는지는 밝혀지지 않았다. 본 연구는 KIF5B의 결합 단백질을 동정하기 위하여 효모 two-hybrid system을 사용하여 KIF5B와 특이적으로 결합하는 Myo9b을 확인하였다. Myo9b는 액틴위를 이동하는 운동단백질로 다른 KIF5s들과도 결합함을 효모 two-hybrid assay로 확인하였다. 또한 Myo9s의 GTPase 활성화 단백질(GAP) 영역은 KIF5B와 결합하는데 필수영역임을 확인하였고, 이러한 단백질간의 결합은 Glutathione S-transferase (GST) pull-down assay를 통하여서도 확인하였다. 생쥐의 뇌 파쇄액에 KIF5B들의 항체로 면역침강을 행하여 Myo9s 단백질을 확인한 결과, KIF5s는 Myo9s 단백질과 특이적으로 함께 침강하였다. 이러한 결과들은 kinesin-I는 액틴 결합 운동단백질과 직접 결합함을 보여준다.

• Clinical and Experimental Reproductive Medicine
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• 제31권2호
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• pp.141-147
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• 2004

Objective: To investigate the association between endometriosis and polymorphisms of N-acetyl transferase 2 (NAT2), glutathione S-transferase M1 (GSTM1), and cytochrome P450 (CYP) 1A1 genes in Korean infertile patients. Materials and Methods: A total of 303 infertile patients who had undertaken diagnostic laparoscopy during January, 2001 through December, 2003 at Samsung Cheil Hospital enrolled in this study. The patients were grouped according to laparoscopic findings: minimal to mild endometriosis (group I: n=147), moderate to severe endometriosis (group II: n=57), normal pelvic cavity (n=99). Peripheral blood was obtained and genomic DNA was extracted. The genotypes of each genes were analyzed using polymerase chain reaction (PCR) and restriction fragment length polymorphism (RFLP). For NAT2, RFLP was used to detect the wild type (wt) and mutant (mt) alleles, enabling classification into slow (mt/mt) or fast (wt/wt or wt/mt) acetylation genotypes. For GSTM1, PCR was used to distinguish active (+/- or +/+) from null (-/-) genotypes. For CYP1A1, MspI digestion was used to detect the wild type (A1A1), heterozygote (A1A2) or mutant (A2A2) genotypes. Result: The genotype frequencies of NAT2 slow acetylator was 12.8%, 10.9%, 12.8% in group I, group II and control, respectively. The genotype frequencies of GSTM1 null mutation was 55.3%, 41.8%, 53.2% in group I, group II and control, respectively. The genotype frequencies of CYP1A1 MspI polymorphism was 16.3%, 9.1%, 18.1% in group I, group II and control, respectively. No significant difference was observed between endometriosis and normal controls in the genotype frequencies of the NAT2, GSTM1, CYP1A1 MspI polymorphism. Conclusion: The NAT2, GSTM1, CYP1A1 gene polymorphism may not be associated with the susceptibility of endometriosis in Korean women.

• 생명과학회지
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• 제14권1호
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• pp.141-147
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• 2004

가자 추출물을 흰쥐에 일정량 투여한 후 간장에 미치는 영향을 관찰할 목적으로 여러 가지 효소량의 변동을 검토하였다. 간 손상의 지표로 사용되는 AST, ALT활성을 측정한 결과 AST, ALT모두에서 정상군보다 300 mg/kg의 용량으로 2주 시료 투여한 군의 함량이 2배 이상 증가되는 것으로 나타났다. 가자 추출물이 지질과산화물 생성에 미치는 효과를 관찰한 결과, 300 mg/kg의 용량으로 2주 시료 투여한 군함량이 정상군에 비해 135.43%, 173.9% 정도 증가하는 결과를 나타내었다. 간 조직의 지질과산화물 반응에 관여하는 것으로 알려진 XO, AO, AD 및 AH활성에 대한 효과를 관찰한 결과 마이크로솜 분획에 존재하는 AD, AH의 활성은 저해 효과가 없었고, 세포질 효소인 XO, AO의 활성은 300 mg/kg의 용량으로 2주 시료 투여한 군 함량이 정상군에 비해 약 2배 증가됨을 볼 수 있었다. 가자 추출물에 의한 간 조직중 glutathione농도에 미치는 영향에 대해 관찰한 결과 2주 동안 300 mg/kg의 용량으로 투여한 군의 glutathione농도는 정상군에 비해 약 75% 감소하는 것으로 나타났다 가자 추출물 투여 후 glutathione의 함량 감소를 경감시키는 기전을 알아볼 목적으로 glutathione 합성에 관여하는 $\gamma$-GCS의 활성과 산화형 glutathione을 환원형 glutathione으로 환원시키는 GR의 활성을 관찰한 결과 정상군에 비해 GR이 56.6%, $\gamma$-GCS가 6.7% 정도 감소하는 것으로 나타났다. 이러한 glutathione의 함량 변동은 산화형 glutathione을 환원형 glutathione으로 환원시키는 GR의 활성에 영향을 주어 나타나는 결과로 생각된다. 가자 추출물이 지질과산화의 해독계에 미치는 영향을 관찰할 목적으로 catalase, GP 및 SOD의 활성을 측정한 결과 catalase, GP의 활성은 각각 77.5%, 64.3% 감소하는 결과를 나타냈으며, SOD의 활성은 정상군에 비해 약 3배 증가하는 것으로 나타났다.

• 한국식품영양과학회지
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• 제20권3호
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• pp.203-208
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• 1991

The present study was undertaken in order to elucidate the effects of pretreatment with nicotinamide on changes in the hepatic metabolizing enzyme system inducted by streptozotocin (STZ). In rats, STZ(50mg/kg) administered by tail vein caused a significant rise in hepatic aniline hydroxylase and a decrease in aminopyrine N-demethylase when compared to control (p<0.05). Pretreatment with nicotinamice inhibited these effects (p<0.05). Similarly, STZ induced changes in hepatic microsomal cytochrome P-450 activity were inhibited by pretreatment with nicotinamide (p<0.05). However, changes in UDP-glucuronyl transferase and sulfortransferase activity were not significantly different(p>0.05). Pretreatment with nicotinamide also prevented STZ induced increases in glutathion S-tranferase activity when compared to the control(p<0.05). There results suggest that nicotinamide pretreatment suppresses STZ-induced changes in the hepatic metabolizing enzyme system.

• Journal of Ginseng Research
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• 제41권3호
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• pp.307-315
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• 2017

Background: Red-skin root disease has seriously decreased the quality and production of Panax ginseng (ginseng). Methods: To explore the disease's origin, comparative analysis was performed in different parts of the plant, particularly the epidermis, cortex, and/or fibrous roots of 5-yr-old healthy and diseased red-skin ginseng. The inorganic element composition, phenolic compound concentration, reactive oxidation system, antioxidant concentrations such as ascorbate and glutathione, activities of enzymes related to phenolic metabolism and oxidation, and antioxidative system particularly the ascorbate-glutathione cycle were examined using conventional methods. Results: Aluminum (Al), iron (Fe), magnesium, and phosphorus were increased, whereas manganese was unchanged and calcium was decreased in the epidermis and fibrous root of red-skin ginseng, which also contained higher levels of phenolic compounds, higher activities of the phenolic compound-synthesizing enzyme phenylalanine ammonia-lyase and the phenolic compound oxidation-related enzymes guaiacol peroxidase and polyphenoloxidase. As the substrate of guaiacol peroxidase, higher levels of $H_2O_2$ and correspondingly higher activities of superoxide dismutase and catalase were found in red-skin ginseng. Increased levels of ascorbate and glutathione; increased activities of $\text\tiny L$-galactose 1-dehydrogenase, ascorbate peroxidase, ascorbic acid oxidase, and glutathione reductase; and lower activities of dehydroascorbate reductase, monodehydroascorbate reductase, and glutathione peroxidase were found in red-skin ginseng. Glutathione-S-transferase activity remained constant. Conclusion: Hence, higher element accumulation, particularly Al and Fe, activated multiple enzymes related to accumulation of phenolic compounds and their oxidation. This might contribute to red-skin symptoms in ginseng. It is proposed that antioxidant and antioxidative enzymes, especially those involved in ascorbate-glutathione cycles, are activated to protect against phenolic compound oxidation.