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  1 / 2020 MEDLINE  
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[PMID]:28398514
[Au] Autor:Strobel EJ; Watters KE; Nedialkov Y; Artsimovitch I; Lucks JB
[Ad] Endereço:Department of Chemical and Biological Engineering, Northwestern University, Evanston, IL 60201, USA.
[Ti] Título:Distributed biotin-streptavidin transcription roadblocks for mapping cotranscriptional RNA folding.
[So] Source:Nucleic Acids Res;45(12):e109, 2017 Jul 07.
[Is] ISSN:1362-4962
[Cp] País de publicação:England
[La] Idioma:eng
[Ab] Resumo:RNA folding during transcription directs an order of folding that can determine RNA structure and function. However, the experimental study of cotranscriptional RNA folding has been limited by the lack of easily approachable methods that can interrogate nascent RNA structure at nucleotide resolution. To address this, we previously developed cotranscriptional selective 2΄-hydroxyl acylation analyzed by primer extension sequencing (SHAPE-Seq) to simultaneously probe all intermediate RNA transcripts during transcription by stalling elongation complexes at catalytically dead EcoRIE111Q roadblocks. While effective, the distribution of elongation complexes using EcoRIE111Q requires laborious PCR using many different oligonucleotides for each sequence analyzed. Here, we improve the broad applicability of cotranscriptional SHAPE-Seq by developing a sequence-independent biotin-streptavidin (SAv) roadblocking strategy that simplifies the preparation of roadblocking DNA templates. We first determine the properties of biotin-SAv roadblocks. We then show that randomly distributed biotin-SAv roadblocks can be used in cotranscriptional SHAPE-Seq experiments to identify the same RNA structural transitions related to a riboswitch decision-making process that we previously identified using EcoRIE111Q. Lastly, we find that EcoRIE111Q maps nascent RNA structure to specific transcript lengths more precisely than biotin-SAv and propose guidelines to leverage the complementary strengths of each transcription roadblock in cotranscriptional SHAPE-Seq.
[Mh] Termos MeSH primário: Biotina/química
Técnicas de Química Analítica
Dobramento de RNA
RNA/química
Estreptavidina/química
Transcrição Genética
[Mh] Termos MeSH secundário: Acilação
Pareamento de Bases
Sequência de Bases
Biotina/genética
Primers do DNA/química
Primers do DNA/genética
Desoxirribonuclease EcoRI/química
Desoxirribonuclease EcoRI/genética
Hidróxidos/química
Conformação de Ácido Nucleico
RNA/biossíntese
RNA/genética
Riboswitch
Análise de Sequência de RNA
Estreptavidina/genética
[Pt] Tipo de publicação:JOURNAL ARTICLE
[Nm] Nome de substância:
0 (DNA Primers); 0 (Hydroxides); 0 (Riboswitch); 63231-63-0 (RNA); 6SO6U10H04 (Biotin); 9013-20-1 (Streptavidin); 9159UV381P (hydroxide ion); EC 3.1.21.- (Deoxyribonuclease EcoRI)
[Em] Mês de entrada:1710
[Cu] Atualização por classe:171011
[Lr] Data última revisão:
171011
[Sb] Subgrupo de revista:IM
[Da] Data de entrada para processamento:170412
[St] Status:MEDLINE
[do] DOI:10.1093/nar/gkx233


  2 / 2020 MEDLINE  
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[PMID]:28126820
[Au] Autor:Camunas-Soler J; Alemany A; Ritort F
[Ad] Endereço:Small Biosystems Lab, Departament de Física de la Matèria Condensada, Facultat de Física, Universitat de Barcelona, Barcelona, Spain.
[Ti] Título:Experimental measurement of binding energy, selectivity, and allostery using fluctuation theorems.
[So] Source:Science;355(6323):412-415, 2017 01 27.
[Is] ISSN:1095-9203
[Cp] País de publicação:United States
[La] Idioma:eng
[Ab] Resumo:Thermodynamic bulk measurements of binding reactions rely on the validity of the law of mass action and the assumption of a dilute solution. Yet, important biological systems such as allosteric ligand-receptor binding, macromolecular crowding, or misfolded molecules may not follow these assumptions and may require a particular reaction model. Here we introduce a fluctuation theorem for ligand binding and an experimental approach using single-molecule force spectroscopy to determine binding energies, selectivity, and allostery of nucleic acids and peptides in a model-independent fashion. A similar approach could be used for proteins. This work extends the use of fluctuation theorems beyond unimolecular folding reactions, bridging the thermodynamics of small systems and the basic laws of chemical equilibrium.
[Mh] Termos MeSH primário: Proteínas de Ligação a DNA/química
Ligantes
Termodinâmica
[Mh] Termos MeSH secundário: Regulação Alostérica
Sítios de Ligação
Desoxirribonuclease EcoRI/química
Equinomicina/química
Ligação Proteica
Imagem Individual de Molécula
[Pt] Tipo de publicação:JOURNAL ARTICLE; RESEARCH SUPPORT, NON-U.S. GOV'T
[Nm] Nome de substância:
0 (DNA-Binding Proteins); 0 (Ligands); 512-64-1 (Echinomycin); EC 3.1.21.- (Deoxyribonuclease EcoRI)
[Em] Mês de entrada:1710
[Cu] Atualização por classe:171120
[Lr] Data última revisão:
171120
[Sb] Subgrupo de revista:IM
[Da] Data de entrada para processamento:170128
[St] Status:MEDLINE
[do] DOI:10.1126/science.aah4077


  3 / 2020 MEDLINE  
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[PMID]:26856634
[Au] Autor:Li Y; Li Y; Wu Y; Lu F; Chen Y; Gao W
[Ad] Endereço:Department of Chemistry and Laboratory for Preparation and Application of Ordered Structural Materials of Guangdong Province, Shantou University, Shantou, Guangdong 515063, PR China.
[Ti] Título:An electrochemiluminescence biosensor for endonuclease EcoRI detection.
[So] Source:Biosens Bioelectron;89(Pt 1):585-591, 2017 Mar 15.
[Is] ISSN:1873-4235
[Cp] País de publicação:England
[La] Idioma:eng
[Ab] Resumo:Endonucleases cleavage of DNA plays an important role in biological and medicinal chemistry. This work was going to develop a reliable and sensitive electrochemiluminescent (ECL) biosensor for detecting endonucleases by using gold nanoparticles graphene composite (GNPs-graphene) as a signal amplifier. Firstly, the GNPs and graphene were simultaneously deposited on the glassy carbon electrode (GCE) by cyclic voltammetry. Then a stem DNA was anchored on the surface of GCE. And with a modifying DNA introduced into the electrode by DNA assembly, a strong ECL signal was obtained. After a DNA modified with ferrocene assembly to the stem DNA, the ECL signal had a sharp decrease due to the quench effect of ferrocene to and the biosensor comes into being a "off" state. With the effect of endonuclease, the ECL signal had a recovery because of the ferrocene being released and the biosensor formed a "on" state. Moreover, the recovery of ECL signal was related to the concentration of endonucleases. Combining specific defined DNA and endonuclease, this method has a potential to detect different endonucleases. In this work, we took the EcoRI as an example to identify the feasibility of ECL biosensor in applying in sensitive detection of endonucleases using a GNPs-graphene signal amplifier. Under optimal condition, the proposed biosensor obtained a low limit of detection (LOD) 5.6×10 UmL . And the stability, selectivity and reproducibility of the biosensor also were researched.
[Mh] Termos MeSH primário: Desoxirribonuclease EcoRI/análise
Técnicas Eletroquímicas/métodos
Ouro/química
Grafite/química
Medições Luminescentes/métodos
Nanoestruturas/química
[Mh] Termos MeSH secundário: Técnicas Biossensoriais/métodos
DNA/química
Compostos Ferrosos/química
Limite de Detecção
Metalocenos
Nanopartículas/química
Nanopartículas/ultraestrutura
Nanoestruturas/ultraestrutura
Reprodutibilidade dos Testes
[Pt] Tipo de publicação:JOURNAL ARTICLE
[Nm] Nome de substância:
0 (Ferrous Compounds); 0 (Metallocenes); 7440-57-5 (Gold); 7782-42-5 (Graphite); 9007-49-2 (DNA); EC 3.1.21.- (Deoxyribonuclease EcoRI); U96PKG90JQ (ferrocene)
[Em] Mês de entrada:1702
[Cu] Atualização por classe:171116
[Lr] Data última revisão:
171116
[Sb] Subgrupo de revista:IM
[Da] Data de entrada para processamento:160210
[St] Status:MEDLINE


  4 / 2020 MEDLINE  
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[PMID]:27992993
[Au] Autor:Naughton BS; Reich NO
[Ad] Endereço:Department of Chemistry and Biochemistry, University of California , Santa Barbara, California 93106, United States.
[Ti] Título:Mechanisms of Protein Translocation on DNA Are Differentially Responsive to Water Activity.
[So] Source:Biochemistry;55(50):6957-6960, 2016 Dec 20.
[Is] ISSN:1520-4995
[Cp] País de publicação:United States
[La] Idioma:eng
[Ab] Resumo:Water plays important but poorly understood roles in the functions of most biomolecules. We are interested in understanding how proteins use diverse search mechanisms to locate specific sites on DNA; here we present a study of the role of closely associated waters in diverse translocation mechanisms. The bacterial DNA adenine methyltransferase, Dam, moves across large segments of DNA using an intersegmental hopping mechanism, relying in part on movement through bulk water. In contrast, other proteins, such as the bacterial restriction endonuclease EcoRI, rely on a sliding mechanism, requiring the protein to stay closely associated with DNA. Here we probed how these two mechanistically distinct proteins respond to well-characterized osmolytes, dimethyl sulfoxide (DMSO), and glycerol. The ability of Dam to move over large segments of DNA is not impacted by either osmolyte, consistent with its minimal reliance on a sliding mechanism. In contrast, EcoRI endonuclease translocation is significantly enhanced by DMSO and inhibited by glycerol, providing further corroboration that these proteins rely on distinct translocation mechanisms. The well-established similar effects of these osmolytes on bulk water, and their differential effects on macromolecule-associated waters, support our results and provide further evidence of the importance of water in interactions between macromolecules and their ligands.
[Mh] Termos MeSH primário: DNA Bacteriano/metabolismo
Desoxirribonuclease EcoRI/metabolismo
Proteínas de Escherichia coli/metabolismo
Osmose/fisiologia
Transporte Proteico/efeitos dos fármacos
DNA Metiltransferases Sítio Específica (Adenina-Específica)/metabolismo
Água/farmacologia
[Mh] Termos MeSH secundário: Sítios de Ligação
Crioprotetores/farmacologia
Metilação de DNA
DNA Bacteriano/química
Desoxirribonuclease EcoRI/química
Dimetil Sulfóxido/farmacologia
Proteínas de Escherichia coli/química
Glicerol/farmacologia
DNA Metiltransferases Sítio Específica (Adenina-Específica)/química
Especificidade por Substrato
[Pt] Tipo de publicação:JOURNAL ARTICLE
[Nm] Nome de substância:
0 (Cryoprotective Agents); 0 (DNA, Bacterial); 0 (Escherichia coli Proteins); 059QF0KO0R (Water); EC 2.1.1.72 (Site-Specific DNA-Methyltransferase (Adenine-Specific)); EC 2.1.1.72 (dam protein, E coli); EC 3.1.21.- (Deoxyribonuclease EcoRI); PDC6A3C0OX (Glycerol); YOW8V9698H (Dimethyl Sulfoxide)
[Em] Mês de entrada:1704
[Cu] Atualização por classe:170427
[Lr] Data última revisão:
170427
[Sb] Subgrupo de revista:IM
[Da] Data de entrada para processamento:161221
[St] Status:MEDLINE


  5 / 2020 MEDLINE  
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[PMID]:27749894
[Au] Autor:Hoppins JJ; Gruber DR; Miears HL; Kiryutin AS; Kasymov RD; Petrova DV; Endutkin AV; Popov AV; Yurkovskaya AV; Fedechkin SO; Brockerman JA; Zharkov DO; Smirnov SL
[Ad] Endereço:Chemistry Department, Western Washington University, Bellingham, WA, United States of America.
[Ti] Título:8-Oxoguanine Affects DNA Backbone Conformation in the EcoRI Recognition Site and Inhibits Its Cleavage by the Enzyme.
[So] Source:PLoS One;11(10):e0164424, 2016.
[Is] ISSN:1932-6203
[Cp] País de publicação:United States
[La] Idioma:eng
[Ab] Resumo:8-oxoguanine is one of the most abundant and impactful oxidative DNA lesions. However, the reasons underlying its effects, especially those not directly explained by the altered base pairing ability, are poorly understood. We report the effect of the lesion on the action of EcoRI, a widely used restriction endonuclease. Introduction of 8-oxoguanine inside, or adjacent to, the GAATTC recognition site embedded within the Drew-Dickerson dodecamer sequence notably reduced the EcoRI activity. Solution NMR revealed that 8-oxoguanine in the DNA duplex causes substantial alterations in the sugar-phosphate backbone conformation, inducing a BI→BII transition. Moreover, molecular dynamics of the complex suggested that 8-oxoguanine, although does not disrupt the sequence-specific contacts formed by the enzyme with DNA, shifts the distribution of BI/BII backbone conformers. Based on our data, we propose that the disruption of enzymatic cleavage can be linked with the altered backbone conformation and dynamics in the free oxidized DNA substrate and, possibly, at the protein-DNA interface.
[Mh] Termos MeSH primário: DNA/metabolismo
Desoxirribonuclease EcoRI/metabolismo
Guanina/análogos & derivados
[Mh] Termos MeSH secundário: Sequência de Bases
Sítios de Ligação
DNA/química
Clivagem do DNA
Dano ao DNA
Guanina/química
Guanina/metabolismo
Cinética
Espectroscopia de Ressonância Magnética
Simulação de Dinâmica Molecular
Conformação de Ácido Nucleico
Estrutura Terciária de Proteína
Especificidade por Substrato
[Pt] Tipo de publicação:JOURNAL ARTICLE
[Nm] Nome de substância:
5614-64-2 (8-hydroxyguanine); 5Z93L87A1R (Guanine); 9007-49-2 (DNA); EC 3.1.21.- (Deoxyribonuclease EcoRI)
[Em] Mês de entrada:1705
[Cu] Atualização por classe:170531
[Lr] Data última revisão:
170531
[Sb] Subgrupo de revista:IM
[Da] Data de entrada para processamento:161018
[St] Status:MEDLINE
[do] DOI:10.1371/journal.pone.0164424


  6 / 2020 MEDLINE  
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[PMID]:26503160
[Au] Autor:Millwood RJ; Moon HS; Poovaiah CR; Muthukumar B; Rice JH; Abercrombie JM; Abercrombie LL; Green WD; Stewart CN
[Ad] Endereço:Department of Plant Sciences, University of Tennessee, Knoxville, TN, USA.
[Ti] Título:Engineered selective plant male sterility through pollen-specific expression of the EcoRI restriction endonuclease.
[So] Source:Plant Biotechnol J;14(5):1281-90, 2016 May.
[Is] ISSN:1467-7652
[Cp] País de publicação:England
[La] Idioma:eng
[Ab] Resumo:Unintended gene flow from transgenic plants via pollen, seed and vegetative propagation is a regulatory concern because of potential admixture in food and crop systems, as well as hybridization and introgression to wild and weedy relatives. Bioconfinement of transgenic pollen would help address some of these concerns and enable transgenic plant production for several crops where gene flow is an issue. Here, we demonstrate the expression of the restriction endonuclease EcoRI under the control of the tomato pollen-specific LAT52 promoter is an effective method for generating selective male sterility in Nicotiana tabacum (tobacco). Of nine transgenic events recovered, four events had very high bioconfinement with tightly controlled EcoRI expression in pollen and negligible-to-no expression other plant tissues. Transgenic plants had normal morphology wherein vegetative growth and reproductivity were similar to nontransgenic controls. In glasshouse experiments, transgenic lines were hand-crossed to both male-sterile and emasculated nontransgenic tobacco varieties. Progeny analysis of 16 000-40 000 seeds per transgenic line demonstrated five lines approached (>99.7%) or attained 100% bioconfinement for one or more generations. Bioconfinement was again demonstrated at or near 100% under field conditions where four transgenic lines were grown in close proximity to male-sterile tobacco, and 900-2100 seeds per male-sterile line were analysed for transgenes. Based upon these results, we conclude EcoRI-driven selective male sterility holds practical potential as a safe and reliable transgene bioconfinement strategy. Given the mechanism of male sterility, this method could be applicable to any plant species.
[Mh] Termos MeSH primário: Infertilidade das Plantas/genética
Tabaco/genética
[Mh] Termos MeSH secundário: Desoxirribonuclease EcoRI/metabolismo
Fluxo Gênico
Engenharia Genética
Hibridização Genética
Especificidade de Órgãos
Plantas Geneticamente Modificadas
Pólen/genética
Regiões Promotoras Genéticas/genética
Sementes/genética
Transgenes
[Pt] Tipo de publicação:JOURNAL ARTICLE; RESEARCH SUPPORT, U.S. GOV'T, NON-P.H.S.
[Nm] Nome de substância:
EC 3.1.21.- (Deoxyribonuclease EcoRI)
[Em] Mês de entrada:1612
[Cu] Atualização por classe:161230
[Lr] Data última revisão:
161230
[Sb] Subgrupo de revista:IM
[Da] Data de entrada para processamento:151028
[St] Status:MEDLINE
[do] DOI:10.1111/pbi.12493


  7 / 2020 MEDLINE  
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[PMID]:26671366
[Au] Autor:Barel I; Reich NO; Brown FL
[Ad] Endereço:Department of Chemistry and Biochemistry, University of California, Santa Barbara, California 93106, USA.
[Ti] Título:Extracting enzyme processivity from kinetic assays.
[So] Source:J Chem Phys;143(22):224115, 2015 Dec 14.
[Is] ISSN:1089-7690
[Cp] País de publicação:United States
[La] Idioma:eng
[Ab] Resumo:A steady-state analysis for the catalytic turnover of molecules containing two substrate sites is presented. A broad class of Markovian dynamic models, motivated by the action of DNA modifying enzymes and the rich variety of translocation mechanisms associated with these systems (e.g., sliding, hopping, intersegmental transfer, etc.), is considered. The modeling suggests an elementary and general method of data analysis, which enables the extraction of the enzyme's processivity directly and unambiguously from experimental data. This analysis is not limited to the initial velocity regime. The predictions are validated both against detailed numerical models and by revisiting published experimental data for EcoRI endonuclease acting on DNA.
[Mh] Termos MeSH primário: DNA/metabolismo
Desoxirribonuclease EcoRI/metabolismo
[Mh] Termos MeSH secundário: DNA/química
Desoxirribonuclease EcoRI/química
Cinética
Cadeias de Markov
[Pt] Tipo de publicação:JOURNAL ARTICLE; RESEARCH SUPPORT, U.S. GOV'T, NON-P.H.S.
[Nm] Nome de substância:
9007-49-2 (DNA); EC 3.1.21.- (Deoxyribonuclease EcoRI)
[Em] Mês de entrada:1609
[Cu] Atualização por classe:151216
[Lr] Data última revisão:
151216
[Sb] Subgrupo de revista:IM
[Da] Data de entrada para processamento:151217
[St] Status:MEDLINE
[do] DOI:10.1063/1.4937155


  8 / 2020 MEDLINE  
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[PMID]:26446158
[Au] Autor:Gu W; Zhang M; Wen S
[Ad] Endereço:Department of Hypertension Research, Beijing Anzhen Hospital, Capital Medical University and Beijing Institute of Heart Lung and Blood Vessel Diseases, 2 Anzhen Road, Beijing, 100029, People's Republic of China.
[Ti] Título:Association between the APOB XbaI and EcoRI polymorphisms and lipids in Chinese: a meta-analysis.
[So] Source:Lipids Health Dis;14:123, 2015 Oct 07.
[Is] ISSN:1476-511X
[Cp] País de publicação:England
[La] Idioma:eng
[Ab] Resumo:BACKGROUND: No previous meta-analysis was to report the association between the apolipoprotein B (APOB) XbaI and EcoRI polymorphisms and serum lipids in Chinese. We performed the study to investigate their potentially association. METHODS AND RESULTS: Studies in English and Chinese were found via a systematic search of Pubmed, Embase, CNKI and Wanfang databases. The dominant genetic model and random-effects model were used to pool data from individual studies. As a result, a total of 30 articles with 5611 subjects for XbaI and 2653 subjects for EcoRI were included in the current study. For the XbaI polymorphism, overall, subjects carrying X+ allele were significantly associated with higher TC,TG and LDL compared with X-X- genotype (Pvalue = 0.0006, OR (95 %) = -0.55 (-0.86,-0.23); Pvalue = 0.0004, OR (95 %) = -0.30 (-0.47,-0.14); (Pvalue = 0.05, OR (95 %) = -0.23(-0.46,-0.00), respectively). Similar results were observed in the subgroups of Han, healthy individuals (HT), coronary heart disease (CHD), cerebral infarction (CI), and cholelithiasis. For HDL, positive association between X+ allele with Lower lipid value was found in CHD and CI subgroups. For EcoRI polymorphism, overall, the E- allele carriers were found to be obviously linked with elevated LDL and lower HDL compared with E + E+ genotype (Pvalue = 0.02,OR (95 %) = -0.27 (-0.49,-0.05); Pvalue = 0.01, OR (95 %) = 0.17 (0.03, 0.30), respectively). TC was significantly high in subjects carrying E- allele in the subgroup of hyperlipidemia. No evidence of publication bias was observed. CONCLUSIONS: The two genetic variants of APOB may be associated with serum lipids in Chinese.
[Mh] Termos MeSH primário: Apolipoproteínas B/genética
Infarto Cerebral/genética
Colelitíase/genética
Doença das Coronárias/genética
Hiperlipidemias/genética
Polimorfismo de Nucleotídeo Único
[Mh] Termos MeSH secundário: Alelos
Apolipoproteínas B/sangue
Grupo com Ancestrais do Continente Asiático
Estudos de Casos e Controles
Infarto Cerebral/sangue
Infarto Cerebral/etnologia
Infarto Cerebral/patologia
Colelitíase/sangue
Colelitíase/etnologia
Colelitíase/patologia
HDL-Colesterol/sangue
LDL-Colesterol/sangue
Doença das Coronárias/sangue
Doença das Coronárias/etnologia
Doença das Coronárias/patologia
Desoxirribonuclease EcoRI/química
Desoxirribonucleases de Sítio Específico do Tipo II/química
Frequência do Gene
Genótipo
Seres Humanos
Hiperlipidemias/sangue
Hiperlipidemias/etnologia
Hiperlipidemias/patologia
Triglicerídeos/sangue
[Pt] Tipo de publicação:JOURNAL ARTICLE; META-ANALYSIS; RESEARCH SUPPORT, NON-U.S. GOV'T
[Nm] Nome de substância:
0 (Apolipoproteins B); 0 (Cholesterol, HDL); 0 (Cholesterol, LDL); 0 (Triglycerides); EC 3.1.21.- (Deoxyribonuclease EcoRI); EC 3.1.21.- (endodeoxyribonuclease XBAI); EC 3.1.21.4 (Deoxyribonucleases, Type II Site-Specific)
[Em] Mês de entrada:1605
[Cu] Atualização por classe:170220
[Lr] Data última revisão:
170220
[Sb] Subgrupo de revista:IM
[Da] Data de entrada para processamento:151009
[St] Status:MEDLINE
[do] DOI:10.1186/s12944-015-0125-z


  9 / 2020 MEDLINE  
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[PMID]:25579883
[Au] Autor:Gani AR; Uppala JK; Ramaiah KV
[Ad] Endereço:Department of Biochemistry, University of Hyderabad, Hyderabad 500046, India.
[Ti] Título:Tauroursodeoxycholic acid prevents stress induced aggregation of proteins in vitro and promotes PERK activation in HepG2 cells.
[So] Source:Arch Biochem Biophys;568:8-15, 2015 Feb 15.
[Is] ISSN:1096-0384
[Cp] País de publicação:United States
[La] Idioma:eng
[Ab] Resumo:Tauroursodeoxycholic acid (TUDCA) a bile salt and chemical chaperone reduces stress-induced aggregation of proteins; activates PERK [PKR (RNA-dependent protein kinase)-like ER (endoplasmic reticulum) kinase] or EIF2AK3, one of the hall marks of ER stress induced unfolded protein response (UPR) in human hepatoblastoma HepG2 cells; prevents heat and dithiothreitol (DTT) induced aggregation of BSA (bovine serum albumin), and reduces ANS (1-anilino-naphthalene-8-sulfonate) bound BSA fluorescence in vitro. TUDCA inactivates heat treated, but not the native EcoR1 enzyme, and reduces heat-induced aggregation and activity of COX-1 (cyclooxygenase enzyme-1) in vitro. These findings suggest that TUDCA binds to the hydrophobic regions of proteins and prevents their subsequent aggregation. This may stabilize unfolded proteins that can mount UPR or facilitate their degradation through cellular degradation pathways.
[Mh] Termos MeSH primário: Ativação Enzimática
Células Hep G2/metabolismo
Agregados Proteicos
Soroalbumina Bovina/metabolismo
Ácido Tauroquenodesoxicólico/metabolismo
eIF-2 Quinase/metabolismo
[Mh] Termos MeSH secundário: Ciclo-Oxigenase 1/metabolismo
Desoxirribonuclease EcoRI/metabolismo
Ditioeritritol/metabolismo
Estresse do Retículo Endoplasmático
Temperatura Alta
Seres Humanos
Resposta a Proteínas não Dobradas
[Pt] Tipo de publicação:JOURNAL ARTICLE; RESEARCH SUPPORT, NON-U.S. GOV'T
[Nm] Nome de substância:
0 (Protein Aggregates); 27432CM55Q (Serum Albumin, Bovine); 516-35-8 (Taurochenodeoxycholic Acid); 60EUX8MN5X (tauroursodeoxycholic acid); 6892-68-8 (Dithioerythritol); EC 1.14.99.1 (Cyclooxygenase 1); EC 1.14.99.1 (PTGS1 protein, human); EC 2.7.11.1 (EIF2AK3 protein, human); EC 2.7.11.1 (eIF-2 Kinase); EC 3.1.21.- (Deoxyribonuclease EcoRI)
[Em] Mês de entrada:1504
[Cu] Atualização por classe:171116
[Lr] Data última revisão:
171116
[Sb] Subgrupo de revista:IM
[Da] Data de entrada para processamento:150113
[St] Status:MEDLINE


  10 / 2020 MEDLINE  
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[PMID]:25463644
[Au] Autor:Zhang S; Hu X; Yang X; Sun Q; Xu X; Liu X; Shen G; Lu J; Shen G; Yu R
[Ad] Endereço:Department of Chemistry and Chemical Engineering, Hunan University of Arts and Science, Changde 415000, PR China; State Key Laboratory for Chemo/Biosensing and Chemometrics, College of Chemistry and Chemical Engineering, Hunan University, Changsha 410082, PR China. Electronic address: zsb0119@163.co
[Ti] Título:Background eliminated signal-on electrochemical aptasensing platform for highly sensitive detection of protein.
[So] Source:Biosens Bioelectron;66:363-9, 2015 Apr 15.
[Is] ISSN:1873-4235
[Cp] País de publicação:England
[La] Idioma:eng
[Ab] Resumo:Using platelet-derived growth factor B chain dimer (PDGF-BB) as the model target, a background current eliminated electrochemical aptameric sensing platform for highly sensitive and signal-on detection of protein is proposed in this paper. Successful fabrication of the biosensor depends on ingenious design of aptamer probe, which contains the aptamer sequence for PDGF-BB and the recognition sequence for EcoRI endonuclease. In the absence of PDGF-BB, the ferrocene labeled aptamer probe folds into a hairpin structure and forms a recognition site for EcoRI. By treatment with endonuclease, the specific and cleavable double-stranded region is cut off and redox-active ferrocene molecule is removed from the electrode surface, and almost no peak current is observed. When binding with target protein, the designed aptamer probe changes its conformation and dissociates the recognition double strand. The integrated aptamer probe is maintained when exposing to EcoRI endonuclease, resulting in obvious peak current. Therefore, a signal-on and sensitive sensing strategy for PDGF-BB detection is fabricated with eliminated background current. Under the optimized experimental conditions, a wide linear response range of 4 orders of magnitude from 20pgmL(-1) to 200ngmL(-1) is achieved with a detection limit of 10pgmL(-1). Moreover, the present aptameric platform is universal for the analysis of a broad range of target molecules of interest by changing and designing the sequence of aptamer probe.
[Mh] Termos MeSH primário: Aptâmeros de Nucleotídeos/química
Técnicas Biossensoriais/métodos
Eletroquímica/métodos
Proteínas Proto-Oncogênicas c-sis/sangue
[Mh] Termos MeSH secundário: Aptâmeros de Nucleotídeos/metabolismo
Sequência de Bases
Desoxirribonuclease EcoRI/metabolismo
Seres Humanos
Limite de Detecção
Proteínas Proto-Oncogênicas c-sis/análise
[Pt] Tipo de publicação:EVALUATION STUDIES; JOURNAL ARTICLE; RESEARCH SUPPORT, NON-U.S. GOV'T
[Nm] Nome de substância:
0 (Aptamers, Nucleotide); 0 (Proto-Oncogene Proteins c-sis); 1B56C968OA (becaplermin); EC 3.1.21.- (Deoxyribonuclease EcoRI)
[Em] Mês de entrada:1508
[Cu] Atualização por classe:161125
[Lr] Data última revisão:
161125
[Sb] Subgrupo de revista:IM
[Da] Data de entrada para processamento:141203
[St] Status:MEDLINE



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