• Journal of Microbiology and Biotechnology
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• 제22권2호
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• pp.176-183
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• 2012

Eukaryotic translation termination is governed by eRF1 and eRF3. eRF1 recognizes the stop codons and then hydrolyzes peptidyl-tRNA. eRF3, which facilitates the termination process, belongs to the GTPase superfamily. In this study, the effect of the MC domain of eRF1a (eRF1aMC) on the GTPase activity of eRF3 was analyzed using fluorescence spectra and high-performance liquid chromatography. The results indicated eRF1aMC promotes the GTPase activity of eRF3, which is similar to the role of eRF1a. Furthermore, the increased affinity of eRF3 for GTP induced by eRF1aMC was dependent on the concentration of $Mg^{2+}$. Changes in the secondary structure of eRF3C after binding GTP/GDP were detected by CD spectroscopy. The results revealed changes of conformation during formation of the eRF3C GTP complex that were detected in the presence of eRF1a or eRF1aMC. The conformations of the eRF3C eRF1a GTP and eRF3C eRF1aMC GTP complexes were further altered upon the addition of $Mg^{2+}$. By contrast, there was no change in the conformation of GTP bound to free eRF3C or the eRF3C eRF1aN complex. These results suggest that alterations in the conformation of GTP bound to eRF3 is dependent on eRF1a and $Mg^{2+}$, whereas the MC domain of eRF1a is responsible for the change in the conformation of GTP bound to eRF3 in Euplotes octocarinatus.

• 약학회지
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• 제49권4호
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• pp.299-305
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• 2005

Structure-activity relationship studies of allylamine type of antimycotics were carried out to evaluate the effect of naphthyl and methyl portion of naftifine. Compounds with 3,4-difluorophenyl (2a-5a), 4-hydroxyphenyl (2b-5b), 3-nitro­phenyl (2c-5c), 4-chlorobenzothiazoly (2d-5d) and 5-methylfurfural (2e-5e) instead of naphthyl group, and with hydrogen (3a-3e), methyl (4a-4e) and ethyl (5a-5e) in the place of methyl in naftifine were synthesized and tested for their in vitro anti-fungal activities against five different fungi. Fourteen compounds (3a, 4a, 5a, 3b, 4b, 5b, 3c, 4c, 5c, 3d, 4d, 3e, 4e and 5e) showed significant anti-fungal activities against T. mentagrophytes. (E)-N-(3-Phenyl-2-propenyl)-3,4-difluoro-ben­zenemethaneamine(3a), (E)_N_(3_phenyl_2_propenyl)_4_hydroxy_benzenemethaneamine(3b) and (E)-N-ethyl-N-(3-phenyl­2-propenyl)-3-nitro-benzenemethaneamine (5c) displayed moderate anti-fungal activities against all five different fungi.

• 공업화학
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• 제20권2호
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• pp.134-139
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• 2009

(E)-3-(Aryl)acrylaldehyde 유도체들 (1a~1e)과 thiosemicarbazide 축합반응으로 일련의 알릴리덴치오세미카르바존 화합물 (2a~2e)을 45~85%로 얻었다. 이 화합물들을 2,4'-dibromoacetophenone와 2,3-dichloroquinoxaline로 처리하여 각각 2,4-이치환 티아졸류(3a~3e)와 2-[(E)-3-(aryl)allylidenehydrazone]thiazolo[5,4-b]quinoxaline류 (4a~4e)를 좋은 수율로 합성하였다. 새로이 합성한 모든 화합물들의 구조들은 IR과 $^1H-NMR$ 분광학 자료로 확인하였다.

• Asian-Australasian Journal of Animal Sciences
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• 제15권3호
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• pp.421-426
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• 2002

The pyruvate dehydrogenase complex (PDC), a member of $\alpha$-keto acid dehydrogenase complex, catalyzes the oxidative decarboxylation of pyruvate with the formation of $CO_2$, acetyl-CoA, NADH, and $H^+$. This complex contains multiple copies of three catalytic components including pyruvate dehydrogenase (E1), dihydrolipoamide acetyltransferase (E2), and dihydrolipoamide dehydrogenase (E3). Two regulatory components (E1-kinase and phospho-E1 phosphatase) and functionally less-understood protein (protein X, E3BP) are also involved in the formation of the complex. In this study, we have partially cloned the gene for E3BP in human. Nine putative clones were isolated by human genomic library screening with 1.35 kb fragment of E3BP cDNA as a probe. For investigation of cloned genes, Southern blot analysis and the construction of the restriction map were performed. One of the isolated clones, E3BP741, has a 3 kb-SacI fragment, which contains 200 bp region matched with E3BP cDNA sequences. The matched DNA sequence encodes the carboxyl-terminal portion of lipoyl-bearing domain and hinge region of human E3BP. Differences between yeast E3BP and mammalian E3BP coupled with the remarkable similarity between mammalian E2 and mammalian E3BP were confirmed from the comparison of the nucleotide sequence and the deduced amino acid sequence in the cloned E3BP. Cloning of human E3BP gene and analysis of the gene structure will facilitate the understanding of the role(s) of E3BP in mammalian PDC.

• 약학회지
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• 제48권4호
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• pp.254-260
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• 2004

Structure-activity relationship studies of allylamine type of antimycotics were carried out to evaluate the effect of naphthyl and methyl portion of naftifine. Compounds with 2,4-difluorophenyl( 2a-5a), 2,5-difluorophenyl(2b-5b), 4-ethylphenyl(2c-5c), 2-hydroxyphenyl(2d-5d) and 2-methylnaphthyl(2e-5e) instead of naphthyl group with hydrogen(3a-3e), methyl(4a-4e), and ethyl(5a-5e) in the place of methyl in naftifine were synthesized and tested their in vitro anti-fungal activity against five different fungi. Eight compounds( 3a, 4a, 5a, 3d, 4d, 4d, 5d, 3e, and 4e) showed significant anti-fungal activity against T. mentagroPhytes. (E)-N-(3-Phenyl-2-propenyl)-2- hydroxy-benzenemethaneamine( 3d) displayed moderate antifungal activity against all five different fungi.

• 한국식품영양과학회지
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• 제28권1호
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• pp.225-232
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• 1999

Apo E polymorphism(e2, e3, e4) was among the first reported genetic polymorphism that explained part of the normal variation in plasma cholesterol concentrations. Among 62 normolipidemic healthy females, aged 19 up to 22 years, the relative frequencies of E3/3 was 0.806(n=50), E3/2 was 0.081(n=5), E3/4 allele was 0.113(n=7), and no E2/2, E2/4 and E4/4 were found. Based on the five samples of E2 allele, five subjects were randomly selected by E3 and E4 groups for the study of effects of apo E polymorphism on the distribution of serum lipid and amino acids profiles. No differences in the anthro pometric data among apo E isomers were found, otherwise the pulsation was higher in E4 than that in the others. There were no differences in plasma total HDL , HDL3 , HDL2 & LDL cholesterol, and apo A I concentrations. However, phenotype means significantly rank E2>E3>E4 allele in average TG levels(p=0.014), and rank E4>E3>E2 in total cholesterol levels(p=0.011). Atherogenic index(AI) such as lipoproteins was significantly increased in E2 & E4 than that in E3(p=0.045). Subjects with E3/2 allele had significantly higher concentrations of glutamine, phosphoserine and taurine, while subjects with E3/4 allele showed significantly lower concentrations of arginine and am inobutyrate and elevated level of phosphoserine in plasma com pared to those of E3/3 allele. Higher level of plasma taurine in subjects with E3/2 or E3/4 allele appears to be related to the elevated level of plasma total and LDL cholesterol concentrations compared to those of E3/3 allele.

• 한국가축번식학회지
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• 제20권2호
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• pp.215-221
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• 1996

This study was carried out to develop an immunoassay for the diagnosis of the pregnancy and ovarian function of domestic animals. Using 2E92C10 monoclonal antibody(McAb) generated against estrone-3-glucuronide(E1-3-G) and appeared a high cross-reactivity with estrone-3-sulfate(E1-3-S), chemiluminesence immunoassay (CIA) to detect E1-3-S was developed. 2E92C10 McAb cross-reacted with E1-3-S (30%) was purified from ascites fluid using protein G sepharose gel column. The purity of purified antibody fraction was monitored by SDS-PAGE and was better compared to that of crude ascite fluid. The soild and liquid phase CIA for E1-3-S were established utilizing 2E92C10 antibody and E1-3-G-ABEI conjugate used as a tracer. As the results, the titer of 2E92C10 antibody was 5g/ml in soild phase and 1:2000 in liquid phase. The sensitivity on soild and solid phase CIA were about 200 pg/ml. These results indicate that CIA for measurement of E1-3-S was successfully developed by using ant-E1-3-G McAb cross-reacted with E1-3-S and could be usefully used to research this area.

• 한국작물학회지
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• 제35권2호
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• pp.146-150
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• 1990

분벽성 옥수수에 대한 esterase 동위효소의 유전양상을 알아보고자 acrylamide gel 6.5%를 사용하여 분벽성인 IK 계통과 대조구로 무벽성인 A-type 계통을 교배하여 얻은 $F_1$ 및 $F_2$ 종자를 28$^{\circ}C$ 암조건하에서 발아시킨 유조직 중심주를 공시하여 전기영동한 결과는 다음과 같다. 1. 본 실험에 공시된 중심주조직은 음극효소를 포함하여 총 13개의 동위효소가 확인되었으며 이동거리에 따라 유전분이 양상이 5개군으로 구분되었다. 2. 원점으로부터 0.5cm에 존재하는 E$_{0.3}$, E$_{0.4}$, E$_{0.5}$ 동위효소는 2개의 대립인자에 의해 지배되었고, E$_{0.3}$+E$_{0.4}$ 변이체는 공우성 대립인자에 의해 지배되었다. 3. 밴드의 이동거리가 1.0cm에서 1.8cm에 존재하는 E$_{1.0}$ , E$_{1.2}$ , E$_{1.5}$ , E$_{1.8}$ 의 4개의 동위효소는 2개의 대립인자에 의해 지배되었고 존재하지 않는 비활성 밴드가 우성으로 작용하였다. 한편 변이체로 나타난 E$_{1.2}$ +E$_{1.5}$ 의 동위효소는 공우성 대립인자로 작용하였다. 4. 밴드의 이동거리가 2.8cm에서 3.5cm에 존재하는 E$_{2.8}$ , E$_{3.0}$ , E$_{3.5}$ 인 세 개의 밴드는 이들 중에서 두 개씩 나타나는 밴드가 세 개 모두 나타나는 개체수보다 현저히 많이 나타났다. ($\chi$$^2$=0.327$^{**}$ ). 5. 밴드의 이동거리가 3.8cm 이상에서 나타난 동위효소는 활성이 매우 낮았고 이들 효소는 모두가 두 개의 대립인자에 의해 지배되었다. 따라서 분벽성 옥수수 중심주 조직에 대한 esterase 동위효소는 멘델식 유전법칙에 의해 지배된다고 생각된다.각된다.각된다.

• Journal of Microbiology and Biotechnology
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• 제20권9호
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• pp.1283-1287
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• 2010

In eukaryotes, eRF3 participates in translation termination and belongs to the superfamily of GTPases. In this work, the dissociation constants for nucleosides bound to Euplotes octocarinatus eRF3 in the presence and absence of eRF1a were determined using fluorescence spectra methods. Furthermore, a GTP hydrolyzing assay of eRF3 was carried out using an HPLC method, and the kinetic parameters for GTP hydrolysis by eRF3 were determined. Consistent with data from humans, the results showed that eRF1a promoted the binding of GTP to eRF3 and the GTP hydrolyzing activity of eRF3. However, in contrast to the lack of GTP binding in the absence of eRF1 in human eRF3, the E. octocarinatus eRF3 was able to bind GTP by itself. The nucleotide binding affinity of the E. octocarinatus eRF3 also differed from the human data. A structure model and amino acid sequence alignment of potential G domains indicated that these differences may be due to valine 317 and glutamate 452 displacing the conserved glycine and lysine involved in GTP binding.

• BMB Reports
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• 제29권2호
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• pp.116-121
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• 1996

In earlier studies, the genes encoding Escherichia coli thioredoxin and Corynebacterium nephridii thioredoxin C-3 were fused via a common restriction site in the nucleotide sequence coding for the active site of the proteins to generate two chimeric thioredoxins, designated E-C3 (N to C-terminal) and C3-E. The hybrid thioredoxins were overexpressed in E. coli from the cloned chimeric thioredoxin genes by a T7 promoter/polymerase system. To investigate the structure-function relationship of thioredoxin, we purified the E-C3 hybrid thioredoxin through ammonium sulfate fractionation, DEAE-cellulose chromatography, and Sephadex G-50 gel filtration. Its purity was examined on SDS-polyacrylamide gel electrophoresis and the molecular weight of the purified E-C3 hybrid thioredoxin was estimated to be 12,000. On native polyacrylamide gels, the purified E-C3 hybrid thioredoxin shows a much lower mobility than E. coli thioredoxin. E-C3 hybrid thioredoxin exhibits a 40-fold lower catalytic efficiency with E. coli thioredoxin reductase than E. coli thioredoxin. It was shown to catalyze the reduction of insulin disulfide by dithiothreitol. The purified E-C3 hybrid thioredoxin was also characterized in other aspects.