• 제목/요약/키워드: thiol

검색결과 426건 처리시간 0.024초

Reactive Oxygen Species and Nitrogen Species Differentially Regulate Neuronal Excitability in Rat Spinal Substantia Gelatinosa Neurons

  • Lee, Hae In;Park, A-Reum;Chun, Sang Woo
    • International Journal of Oral Biology
    • /
    • 제39권4호
    • /
    • pp.229-236
    • /
    • 2014
  • Reactive oxygen species (ROS) and nitrogen species (RNS) are implicated in cellular signaling processes and as a cause of oxidative stress. Recent studies indicate that ROS and RNS are important signaling molecules involved in nociceptive transmission. Xanthine oxidase (XO) system is a well-known system for superoxide anions ($O{_2}^{{\cdot}_-}$) generation, and sodium nitroprusside (SNP) is a representative nitric oxide (NO) donor. Patch clamp recording in spinal slices was used to investigate the role of $O{_2}^{{\cdot}_-}$ and NO on substantia gelatinosa (SG) neuronal excitability. Application of xanthine and xanthine oxidase (X/XO) compound induced membrane depolarization. Low concentration SNP ($10{\mu}M$) induced depolarization of the membrane, whereas high concentration SNP (1 mM) evoked membrane hyperpolarization. These responses were significantly decreased by pretreatment with phenyl N-tert-butylnitrone (PBN; nonspecific ROS and RNS scavenger). Addition of thapsigargin to an external calcium free solution for blocking synaptic transmission, led to significantly decreased X/XO-induced responses. Additionally, X/XO and SNP-induced responses were unchanged in the presence of intracellular applied PBN, indicative of the involvement of presynaptic action. Inclusion of GDP-${\beta}$-S or suramin (G protein inhibitors) in the patch pipette decreased SNP-induced responses, whereas it failed to decrease X/XO-induced responses. Pretreatment with n-ethylmaleimide (NEM; thiol-alkylating agent) decreased the effects of SNP, suggesting that these responses were mediated by direct oxidation of channel protein, whereas X/XO-induced responses were unchanged. These data suggested that ROS and RNS play distinct roles in the regulation of the membrane excitability of SG neurons related to the pain transmission.

Bacillus stearothermophilus 에서 부분 정제한 Cytosine Deaminase 의 특성

  • 장영채;이경형;김성영;조윤래;김종규
    • 미생물학회지
    • /
    • 제30권4호
    • /
    • pp.305-309
    • /
    • 1992
  • Bacillus stearothermophilus 의 cytosine deaminase (EC 3. 5. 4.1 )를 수율 52.7% 로 7.2배 부분정제했다. 부분정제된 cytosine deaminase 는 cytosine 을 유일한 기질로 이용하였으머 5-methylcytosine 과 5-fluorocytosine 은 기질고 이용되지 못햇으며 cytosine 에 대한 Michaelis 정수 Km 값은 5.9 mM 이었다. 본효소는 pH 4.0 에서 7.0 까지의 폭 넓은 pH 영역에서 안정했으며 80.deg.C 에서 10 분간 열처리하여도 75% 이상의 효소활성이 잔존하여 높은 내열성 효소였다. 본 효소의 반응최적 pH 는 7.0-7.5 였으며 반응 최적 온도는 35. 37.deg.C 였다. 그리고 Arrhenium plot 에 의하여 계산된 활성화 에너지 값(Ea value) 은 26 Kcal/mol 이었다. 본 효소는 중금속인 1 mM의 $Cd^{2+}$ , $Hg^{2+}$$Cu^{2+}$ 에 의하여 완정히 실활되었으며 GMP 와 CMP 에 의해서는 효소활성이 촉진되었다. 특히 본 효소는 p-chloromercuribenzoate 에 의하여 효소 활성이 강하게 저해되어 thiol 효소임을 추정할 수 있었다.

  • PDF

Effect of Chelation with Calcium Disodium EDTA on Haemato-biochemical and Trace Mineral Profile in Blood from Lead Exposed Calves

  • Patra, R.C.;Swarup, D.
    • Asian-Australasian Journal of Animal Sciences
    • /
    • 제18권8호
    • /
    • pp.1130-1134
    • /
    • 2005
  • An experiment was performed using 20 calves of about one-month old to investigate the effect of chelation therapy with calcium disodium ethylenediaminetetraacetate ($CaNa_2$EDTA) alone or along with antioxidant $\alpha$-tocopherol in lead loaded calves on blood trace minerals, erythrocytic sulfahydryl groups and some haematobiochemical parameters. Fifteen calves were given lead orally at a daily dose of 7.5 mg of 99% pure lead acetate/kg body weight for 28 days. Then the lead was withdrawn on day 28 and the calves were randomly divided into three groups. Each group of five animals was either treated with $CaNa_2$EDTA alone at the dose rate of 110 mg/kg body weight in two divided doses for 4 days or along with $\alpha$-tocopherol at the dose rate of 100 mg/kg body weight orally daily for 7 days, keeping the remaining five calves as lead-exposed untreated controls. Blood samples were collected at the end of the lead exposure (day 0) and thereafter on day 2, 4, 7 and 10 from the start of the chelation treatment. The treatment with EDTA alone led to slow but non-significant improvement in blood copper level, but incorporation of antioxidant $\alpha$-tocopherol in chelation therapy resulted in its significant decline, as recorded on day 7-post treatment. Withdrawal of lead or treatment with $CaNa_2$EDTA alone or along with $\alpha$-tocopherol enhanced the erythrocytic thiol contents and the levels of T-SH and P-SH became statistically (p<0.05) comparable to those of lead-exposed controls by day 7 and 4, respectively. There was no significant (p>0.05) change in serum urea, creatinine, total protein and albumin levels between the treatment groups. It is concluded from the present investigation that treatment with $CaNa_2$EDTA at the present dose rate is safe to be used for chelation in lead loaded calves.

Antimicrobial Effect of 2-Phenylethynyl-Butyltellurium in Escherichia coli and Its Association with Oxidative Stress

  • Pinheiro, Franciane Cabral;Bortolotto, Vandreza Cardoso;Araujo, Stifani Machado;Poetini, Marcia Rosula;Sehn, Carla Pohl;Neto, Jose S.S.;Zeni, Gilson;Prigol, Marina
    • Journal of Microbiology and Biotechnology
    • /
    • 제28권7호
    • /
    • pp.1209-1216
    • /
    • 2018
  • This study aimed to evaluate the antimicrobial activity of 2-phenylethynyl-butyltellurium (PEBT) in Escherichia coli and the relation to its pro-oxidant effect. For this, we carried out the disk diffusion test, minimum inhibitory concentration (MIC) assay, and survival curve analysis. We also measured the level of extracellular reactive oxygen species (ROS), activity of the antioxidant enzymes superoxide dismutase (SOD) and catalase (CAT), and level of non-protein thiols (NPSH). PEBT at 1.28 and 0.128 mg/disk exhibited antimicrobial capability in the disk diffusion test, with an MIC value of 1.92 mg/ml, whereas PEBT at 0.96, 1.92, and 3.84 mg/ml inhibited bacterial growth after a 9-h exposure. PEBT at 3.84, 1.92, and 0.96 mg/ml increased extracellular ROS production, decreased the intracellular NPSH level, and reduced the SOD and CAT activities. Glutathione or ascorbic acid in the medium protected the bacterial cells from the antimicrobial effect of PEBT. In conclusion, PEBT exhibited antimicrobial activity against E. coli, involving the generation of ROS, oxidation of NPSH, and reduction of the antioxidant defenses in the bacterial cells.

가자(Terminalia chebula) 추출물이 마우스의 생리활성에 미치는 영향 (Effect of Terminalia chebula on Physiological Activity in Mice)

  • 박종옥;이승은
    • 생명과학회지
    • /
    • 제14권1호
    • /
    • pp.148-153
    • /
    • 2004
  • 가자 추출물을 취하여 사람, 돼지, 쥐 및 개 등의 3% 적혈구용액으로 응집력시험을 행한 결과 7가지 적혈구 모두에 렉틴 활성이 나타났으며 $LD_{50}$는 390 mg/kg으로 측정되었다. 생리 활성에 대한 영향을 알아보고자 생체 내외인성 요인에 의한 친전자성 물질로 생체내에서 독작용, 노화, 발암 및 면역 억제작용을 유발하는 원인 물질인 free radical생성에 관여하는 효소인 XO 및 AO의 활성을 측정 한 결과, XO 에서는 일주일간 매일 300 mg/kg의 용량으로 가자 추출물을 투여한 군이 정상군보다 5배 증가되는 결과를 나타내었고 AO에서는 정상군보다 시료 투여군이 2배 증가되는 결과를 나타내었다 glutathione은 단백질이나 DNA합성, amino acid의 이동 반응 및 thiol기의 저장 등과 같은 생물학적으로 중요한 여러 가지 반응에 직접 관여하는 물질이다[16〕. 이에 간장 조직의 glutathione농도를 측정한 결과 간장 독성의 유발로 인한 효소 활성은 정상군에 비하여 1주간 매일 300 mg/kg의 용량으로 시료 투여한 군의 효소 활성이 79% 감소됨을 볼 수 있었다. 체내의 여러 가지 해독반응과정에 관여하는 GST효소활성을 측정 한 결과 정상군에 비해 1주간 매 일 300 mg/kg의 용량으로 시료 투여한 군이 66%정도 감소된다는 결과를 볼 수 있었다. glutathione 합성에 관여하는 $\gamma$-GCS의 활성과 산화형 glutathione을 환원형 glutathione으로 환원시키는 GR의 활성을 관찰한 결과 가자 추출물 투여군이 정상군보다 GR의 활성은 80% 감소되었고, 합성에 관여하는 $\gamma$-GCS의 활성은 정상군과 비교할 때 약간의 감소만을 나타내 glutathione함량 변동에 크게 영향을 미치지 않는 것으로 생각된다.

Carbamate-Based Surface Reactions for Release of Amine Molecules from Electroactive Self-Assembled Monolayers

  • Hong, Dae-Wha;Kang, Kyung-Tae;Hong, Seok-Pyo;Shon, Hyun-Kyong;Lee, Tae-Geol;Choi, In-Sung S.
    • 한국진공학회:학술대회논문집
    • /
    • 한국진공학회 2011년도 제41회 하계 정기 학술대회 초록집
    • /
    • pp.208-209
    • /
    • 2011
  • In this work, we developed self-assembled monolayers (SAMs) of alkanethiols on gold that can release amine groups, when an electrical potential was applied to the gold. The strategy was based on the introduction of the electroactive carbamate group, which underwent the two-electron oxidation with simultaneous release of the amine molecules, to alkanethiols. The synthesis of the designed thiol compounds was achieved by coupling isocyanate-containing compound with hydroquinone. The electroactive thiols were mixed with hydroxyl-containing alkanethiol [$HS(CH_2)_{11}OH$] to form mixed monolayers, and cyclic votammetry was used for the characterization of the release. The mixed SAMs showed a first oxidation peak at +540 mV (versus Ag/AgCl reference electrode), demonstrating irreversible conversion from carbamate to hydroqinone with simultaneous release of the amine groups. The second and third cycles showed typical reversible redox reaction of hydroquinone and quione: the oxidation and reduction occurred at +290 mV and -110 mV, respectively. The measurement of ToF-SIMS further indicates that electrochemical-assisted chemical reaction successfully released amine groups. This new SAM-based electrochemistry would be applicable for direct release of biologically active molecules that contain amine groups.

  • PDF

Glutathione (GSH) Response as a Metabolic Biomarker to Benzo(α)pyrene and Aroclor 1254 Exposure in the Pacific Oyster Crassostrea gigas

  • Jo, Qtae;Choy, Eun-Jung;Lee, Yong-Hwa;Ko, Sung-Jung;Song, Young-Chae;Kim, Myoung-Jin;Hwang, Eung-Ju
    • Fisheries and Aquatic Sciences
    • /
    • 제9권4호
    • /
    • pp.140-145
    • /
    • 2006
  • We measured activities of the ubiquitous tripeptide non-protein thiol (L-${\gamma}$-glutamyl-L-cysteinyl-glycine), glutathione (GSH), which is believed to playa fundamental role in detoxifying xenobiotics in biological systems, as a metabolic biomarker for benzo(${\alpha}$)pyrene and Aroclor 1254 exposure in the Pacific oyster Crassostrea gigas. Reproductive oysters were exposed to the pollutants for 50 days by the algal vectoring method in which the oysters were fed with concentrated standard algal foods grown in culture media containing Aroclor 1254 (0, 5, 50, 500 ng/g) or benzo(${\alpha}$)pyrene (0, 10, 100, 1,000 ng/g). Both pollutants induced maternal GSH activities in 10 days in a dosage-dependent manner (p<0.05), although Aroclor 1254 was stronger. The pollutant-driven GSH elevation persisted for 20 to 30 days depending on the pollutants and concentrations. Thereafter, a drastic decline in the GSH activity was observed due to metabolic failure, after which the oyster GSH remained at low levels throughout the remainder of the experiment. The pollutant exposures influenced maternal reproductive output in terms of fertilization, hatching, and morphology. These results imply that changes in activity of the GST-catalyzing molecule can be used as an oyster biomarker for Aroclor 1254 and benzo(${\alpha}$)pyrene exposure.

Quantitative Mass Spectrometric Analysis of Mixed Self-Assembled Monolayers for Biochips

  • Son, Jin Gyeong;Shon, Hyun Kyong;Hong, Daewha;Choi, Changrok;Han, Sang Woo;Choi, Insung S.;Lee, Tae Geol
    • 한국진공학회:학술대회논문집
    • /
    • 한국진공학회 2013년도 제44회 동계 정기학술대회 초록집
    • /
    • pp.275-275
    • /
    • 2013
  • Formation and characterization of self-assembled monolayers (SAMs) on various surfaces are the essential basis for many other applications, including molecular switches, biosensors, microfluidics, and fundamental studies in surfaces and interfaces. To improve the performance at these applications, it is a key to control the quantity of each molecule in various mixed SAMs on the surface. In this study, using mixed SAM of carbamate-based hydroquinone (HQ)-PhBr and11-mercaptoundecanol, the quantitative mass spectrometric method of mixed SAM was developed based on comparison study with XPS and FT-IR methods. In addition, our method was applied to another mixed SAM of biotinylated PEG alkane thiol and 11-mercaptoundecanol for verification purpose. Time-of-flight secondary mass spectrometry (ToF-SIMS) analysis was performed to identify and quantify each molecule of mixed SAM along with principal component analysis (PCA). Since there is no matrix effect in the X-ray photoelectron spectroscopy (XPS) and Fourier transform-infrared (FT-IR) techniques, we compared ToF-SIMS results with XPS and FT-IR results. Because PCA results from ToF-SIMS analysis are well matched with XPS and FT-IR results from both mixed SAMs, we are expecting that our method will be useful to identify and quantify each molecule in various mixed SAMs.

  • PDF

Exploration of the Glycosyltransferase BmmGT1 from a Marine-Derived Bacillus Strain as a Potential Enzyme Tool for Compound Glycol-Diversification

  • Liu, Quanquan;Ren, Pengfei;Liu, Yang;Qin, Wen;Li, Huayue;Li, Wenli
    • Journal of Microbiology and Biotechnology
    • /
    • 제28권6호
    • /
    • pp.931-937
    • /
    • 2018
  • Glycosyltransferases (GTs) from microbes are an emerging and rich source for efficient glycol-transformation of natural/unnatural compounds. Here, we probed the catalytic capability and substrate promiscuity of BmmGT1 from marine-derived Bacillus methylotrophicus B-9987. The regioselectivity of BmmGT1 on macrolactin A (1) was explored by optimization of the reaction conditions, in which a series of O-glycosylated macrolactins (1a-1e) were generated, including two new di/tri-O-glucosyl analogs (1b and 1e). Furthermore, BmmGT1 was able to catalyze the glycosylation of the thiol (S-) or amine (N-) sites of phenolic compounds (2 and 3), leading to the generation of N- (2a) or S-glycosides (3a and 3b). The present study demonstrates that BmmGT1 could serve as a potential enzyme tool for O-, N-, or S-glycosyl structural diversification of compounds for drug discovery.

카드뮴이 뇌혈관 내피세포에서의 $PGE^2$ 및 COX-2 발현에 미치는 영향 (Cadmium-induced COX-2 Expression in Cerebrovascular Endothelial Cells)

  • 박동현;김영채;문창규;정이숙;백은주;문창현;이수환
    • Environmental Analysis Health and Toxicology
    • /
    • 제21권3호
    • /
    • pp.275-282
    • /
    • 2006
  • In order to get insight into the mechanism of cadmium (Cd)-induced brain injury, we investigated the effects of Cd on the induction of COX-2 in bEnd.3 mouse brain endothelial cells. Cd induced COX-2 expression and $PGE_2$ release, which were attenuated by thiol-reducing antioxidant N-acetylcysteine (NAC) indicating oxidative components might contribute to these events. Indeed, Cd increased cellular reactive oxygen species (ROS) level and DNA binding activity of nuclear factor-kB (NF-kB), an oxidative stress sensitive transcription factor. Cd-induced $PGE_2$ production and COX-2 expression were significantly attenuated by Bay 11 7082, a specific inhibitor of NF-kB and by SB203580, a specific inhibitor of p38 mitogen activated protein kinase (MAPK). These data suggest that Cd induces COX-2 expression through activation of NF-kB and p38 MAPK, the oxidative stress-sensitive signaling molecules, in brain endothelial cells.