• 한국동물학회지
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• 제25권3호
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• pp.115-122
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• 1982

소의 혈장으로부터 알부민을 순수 정제하였으며 순도는 면역 화학적 방법을 사용하여 확인하였다. 정제된 알부민에 maleate, iodoacetate, iodoacetamide 및 glutathione의 4가지 thiol 화합물을 각각 반응시켜 그 복합체를 9가지의 상이한 완충용액내에서 초산셀룰로우즈 전기영동한 결과 barbital buffer와 Na-acetate buffer를 제외한 다른 완충용액내에서 albumin-glutathione 복합체는 두 가지의 단백질로 분리되었으며 pH 4.8 citrate buffer 및 pH 4.8 succinate buffer내에서 albumin-iodoacetate 및 albumin-iodoacetamide 복합체는 두 가지의 단백질로 분리되었다. 전기영동상에 나타나는 알부민 분획에는 conformation이 서로 다른 두 가지 이상의 알부민 분자가 존재한다고 사료된다.

• Archives of Pharmacal Research
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• 제26권12호
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• pp.1047-1054
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• 2003

Molecular chaperones have a crucial role in the folding of nascent polypeptides in endoplasmic reticulum. Some of them are known to be sensitive to the modification by electrophilic metabolites of organic pro-toxicants. In order to identify chaperone proteins sensitive to alkyators, ER extract was subjected to alkylation by 4-acetamido-4 -maleimidyl-stilbene-2,2 -disulfonate (AMS), and subsequent SDS-PAGE analyses. Protein spots, with molecular mass of 160, 100, 57 and 36 kDa, were found to be sensitive to AMS alkylation, and one abundant chaperon protein was identified to be protein disulfide isomerase (PDI) in comparison with the purified PDI. To see the reactivity of PDI with cysteine alkylators, the reduced form ($PDI_{red}$) of PDI was incubated with various alkylators containing Michael acceptor structure for 30 min at $38^{\circ}C$ at pH 6.3, and the remaining activity was determined by the insulin reduction assay. Iodoacetamide or N-ethylmaleimide at 0.1 mM remarkably inactivated $PDI_{red}$ with N-ethylmaleimide being more potent than iodoacetamide. A partial inactivation of $PDI_{oxid}$ was expressed by iodoacetamide, but not N-ethylmaleimide (NEM) at pH 6.3. Of Michael acceptor compounds tested, 1,4-benzoquinone ($IC_{50}, 15 \mu$ M) was the most potent, followed by 4-hydroxy-2-nonenal and 1,4-naphthoquinone. In contrast, 1,2-naphthoquinone, devoid of a remarkable inactivation action, was effective to cause the oxidative conversion of $PDI_{red}$ to $PDI_{oxid}$. Thus, the action of Michael acceptor compounds differed greatly depending on their structure. Based on these, it is proposed that POI, one of chaperone proteins in ER, could be susceptible to endogenous or xenobiotic Michael acceptor compounds in vivo system.

• 동의생리병리학회지
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• 제21권6호
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• pp.1549-1554
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• 2007

To verify the effects of JAUN-1, which is a water-extracted herbal mixture, on gastroenteric disorders induced by 0.1 percent of iodoacetamide (IA) in rats. We divided four groups, $Na{\"{\i}}ve$ + Distilled Water (DW), 0.1% IA + DW, 0.1% IA + Proton pump inhibitor (Lansoprazole, 5 mg/kg) and 0.1% IA + Herbal mixture (JAUN-1, 50mg/kg) and performed following experimental methods to confirm its advantageous effects against ulcerogenic stomach in rats induced by 0.1% IA; cell cytotoxicity, analysis of lesions score, Hematoxylin & Eosin (H&E) stain, RT-PCR for ${\beta}-actin$, COX-1 and COX-2 and evaluation of intestinal prokinetic activity. No cytotoxicity was elucidated at the concentration of 1, 5, 10, 50, 100, $500\;{\mu}g/ml$ and 1mg/ml JAUN-1 through MTT Assay using by human stomach epithelial AGS cells, respectively. In addition, the JAUN-1 treated group and the lansoprazole treated group significantly decreased in lesions score compared to the DW treated group in the gastritis induced rat model, and results of immunohistochemistry by H&E staining showed that histological recovery in Proton Pump Inhibitor (PPI) and JAUN-1 treated groups rather than the DW administrated group. Another outcome was that ${\beta}-actin$ relative COX-2 expression level was significantly promoted in the DW treated group while ${\beta}-actin$ relative COX-1 expression level was no meaningful change in this rat model. Finally, intestinal prokinetic activity was recovered from low level of prokinetic activity due to 0.1% IA induced gastritis to the similar level of Normal group. These results suggested that JAUN-1 may have beneficial effects against 0.1% IA-induced gastritis rat model through decreasing lesions score, histological recovery, ${\beta}-actin$ relative COX-1, 2 expression level and prokinetic activity.

• 미생물학회지
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• 제27권3호
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• pp.237-244
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• 1989

Azocasein을 기질로 사용하여 nutrient broth에서 성장한 Pseudomonas carboxydovorans로부터 다섯 단계의 순화 과정을 거쳐 68배 순화된 세포내 가용성 단백질 가수 분해 효소를 얻었다. Pyruvate나 succinate, acetate, 또는 일산화탄소를 이용하여 성장한 세균들은 이 효소의 활성을 나타내지 않았다. 순화된 효소의 크기는 약 53,000이었고, 한개의 polypeptide로 구성되어 있었다. 이 효소는 serinerp통의 단백질 가수분해 효소로 $Cd^{2+}, Cu^{2+}, Hg^{2+}$, 등의 2가 양이온과 EGTA에 의해 활성이 완전히 억제되었고 iodoacetamide에 의해 활성이 증가되었다. 이 효소는 pH 8.0과 $50^{\circ}C$에서 최대 활성을 나타내었으며, 일산화탄소 산화효소를 가수분해 시키지 못하였다.

• The Korean Journal of Physiology and Pharmacology
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• 제17권5호
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• pp.469-477
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• 2013

This study investigated effect of extract containing quercetin-3-O-${\beta}$-D-glucuronopyranoside from Rumex Aquaticus Herba (ECQ) against chronic gastritis in rats. To produce chronic gastritis, the animals received a daily intra-gastric administration of 0.1 ml of 0.15% iodoacetamide (IA) solution for 7 days. Daily exposure of the gastric mucosa to IA induced both gastric lesions and significant reductions of body weight and food and water intake. These reductions recovered with treatment with ECQ for 7 days. ECQ significantly inhibited the elevation of the malondialdehyde levels and myeloperoxidase activity, which were used as indices of lipid peroxidation and neutrophil infiltration. ECQ recovered the level of glutathione, activity of superoxide dismutase (SOD), and expression of SOD-2. The increased levels of total NO concentration and iNOS expression in the IA-induced chronic gastritis were significantly reduced by treatment with ECQ. These results suggest that the ECQ has a therapeutic effect on chronic gastritis in rats by inhibitory actions on neutrophil infiltration, lipid peroxidation and various steps of reactive oxygen species (ROS) generation.

• 한국균학회지
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• 제24권4호통권79호
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• pp.255-264
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• 1996

호알카리성 진균 Cephalosporium sp. RYM-202가 생산하는 alkaline xylanase (CX-III)의 작용에 의해 xylan 기질로부터 생성되는 주요 가수분해 산물은 xylobiose와 중합도가 4이상인 xylooligosaccharides이었다. 이 효소는 xylobiose에 대한 분해능을 가지고 있지 않지만 xylotriose로부터는 xylobiose를, xlyotetraose로부터 xylobiose와 xylotriose를 주산물로 형성하였다. 이러한 결과들은 CX-III가 transglycosidase 활성을 소유하는 전형적인 endo-type xylanase임을 보여준다. N-bromosuccinimide에 의한 CX-III의 화학적 변환 실험결과 효소 1분자 당 2개의 tryptophan 잔기가 활성에 관여하는 것으로 나타났다. 그러나 iodoacetamide 및 diethylpyrocarbonate에 의한 효소활성의 저해효과는 나타나지 않음으로써 이 효소의 활성부위에 cysteine과 histidine 잔기가 필수적이지 않음이 확인되었다.

• 미생물학회지
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• 제32권4호
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• pp.306-314
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• 1994

Xylan과 관련 다당류 (xylotriose, xylotetraose, arabinoxylotriose)에 대한 Trichoderma koningii ATCC 26113에서 분리된 xylanase II의 작용양상은 xylanase II가 endo-enzyme이고 transxylosidation의 활성을 가지고 있다고 보여진다. Xylanase II에 의해 형성된 반응산물을 $^1HNMR$ 분광법으로 분석한 결과는 본 효소에 의해 얻어진 xylooligosaccharides의 가수분해산물은 모두가 ${\beta}$-1,4-xylosidic linkage만을 가지고 있는 것으로 판명되었다. 본 효소를 iodoacetamide로 화학적으로 변형시켰을 때 효소 mole당 cysteine 잔기가 두 개가 활성에 필요한 것으로 보여졌으며, N-bromosuccinimide 로 처리하였을 때는 활성부위에 tryptophan 잔기가 여덟 개 존재하는 것으로 판명되었다.

• BMB Reports
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• 제30권2호
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• pp.157-161
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• 1997

The present paper reports characteristics and specificity of the inhibitory action of $N^{\alpha}-tosyl-L-lysine-chloromethyl\;ketone$ (TLCK) and $N^{\alpha}-tosyl-L-phenylalanine-chloromethyl\;ketone$ (TPCK) on the glucose6-phosphate transporter of rat liver microsomes. The TLCK-induced inhibition was pH dependent. The inhibition constants for TPCK were determined by following pseudo-Lst order reaction mechanism. The inhibition was protected by preincubation with excess amount of glucose-6-phosphate. The results proved that (a) TLCK inactivates the microsomal glucose-6-phosphate transporter, (b) the inhibition results from the modification of sulfhydryl groups of the transporter.

• Journal of Microbiology and Biotechnology
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• 제19권9호
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• pp.918-921
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• 2009

In the present study, we examined the inhibitory effects of protein tyrosine phosphatase (PTPase) inhibitors, including sodium orthovanadate (SOV), ammonium molybdate (AM), and iodoacetamide (IA), on cell growth, accumulation of astaxanthin, and PTPase activity in the photosynthetic algae Haematococcus lacustris. PTPase activity was assayed spectrophotometrically and was found to be inhibited 60% to 90% after treatment with the inhibitors. SOY markedly abolished PTPase activity, significantly activating the accumulation of astaxanthin. These data suggest that the accumulation of astaxanthin in H. lacustris results from the concerted actions of several PTPases.

• 미생물학회지
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• 제30권4호
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• pp.299-304
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• 1992

An intracellular protease form cells of Pseudomonas carboxydovorans DSM 1227 grown on carbon monoxide was purified 57-fold in six steps to homogeneity with a yield of 4.3% using azocoll as a substrate. The molecular weight of the enzyme was determined to be 150,000. Sodium dodecyl sulfate-gel electrophoresis revealed the purified enzyme to be a dimer with two identical subunits of molecular weight 72,000. The enzyme was stimulated by $Mg^{2+}$ but was inhibited completely by $Cd^{2+}$ $Fe^{2+}$ $Hg^{2+}$, and $^Zn{2+}$ The enzyme activity was also inhibited by EDTA, EGTA, phenylmethylsulfonyl fluoride, and phenyl glyoxal, but was increased by 1-ethyl-3(dimethyl aminopropyl fluoride, and phenyl glyoxal, but was increased by 1-ethyl-3(dimethyl aminopropyl)carbodiimide, iodoacetamide and dithiothereitol. The optimal pH and temperature for the enzyme reaction were found to be 7-8 and 50.deg.C, respectively. Casein and bovine serum albumin were hydrolyzed by the enzyme, but carbon monoxide dehydrogenase was not.