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  1 / 4372 MEDLINE  
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[PMID]:29216587
[Au] Autor:Voitechovic E; Korepanov A; Kirsanov D; Legin A
[Ad] Endereço:St. Petersburg State University, St. Petersburg, Russia; Institute of Microelectronics of Barcelona, Barcelona, Spain. Electronic address: voitechovic.edita@gmail.com.
[Ti] Título:Quantification of immobilized protein in pharmaceutical production by bio-assisted potentiometric multisensor system.
[So] Source:J Pharm Biomed Anal;150:67-71, 2018 Feb 20.
[Is] ISSN:1873-264X
[Cp] País de publicação:England
[La] Idioma:eng
[Ab] Resumo:Quantification of proteins is a key biochemical assay in molecular biology, biotechnology, medicine and pharmacology. Protein quantification protocols can be based on spectrophotometry, enzyme-linked immunosorbent assay, mass spectrometry or quantitative immunoblotting depending on analyte. In case of immobilized protein these methods require suitable sample preparation. Thus, sophisticated analysis becomes even more complex, expensive and time-consuming. Such drawbacks are highly undesirable in industry. In this study we propose a new approach for evaluation of immobilized protein concentration based on application of bio-assisted potentiometric multisensor system. Surface-immobilized recombinant protein A from Staphylococcus aureus (SpA, expressed in Escherichia coli), which is commonly used as affinity ligand immobilized to stationary phase (сhromatography media) for monoclonal antibody purification was employed as the model object. Chromatography media samples containing different amounts of immobilized SpA were analyzed. Proteinase K from Tritirachium album was employed as a bio-transducer. We demonstrated that the suggested approach provides information about immobilized SpA concentration with 0.8mg/ml accuracy in the range 1-6.7mg/ml and within just 16min. Moreover, the proposed procedure requires no expensive materials and equipment and no bio-transducer immobilization. This method has potential of application for fast monitoring of other immobilized proteins in different tasks.
[Mh] Termos MeSH primário: Ensaio de Imunoadsorção Enzimática/métodos
Proteínas Imobilizadas/química
Potenciometria/métodos
Proteína Estafilocócica A/química
[Mh] Termos MeSH secundário: Endopeptidase K/química
Proteínas Imobilizadas/análise
Proteínas Recombinantes/análise
Proteínas Recombinantes/química
Reprodutibilidade dos Testes
Proteína Estafilocócica A/análise
[Pt] Tipo de publicação:JOURNAL ARTICLE
[Nm] Nome de substância:
0 (Immobilized Proteins); 0 (Recombinant Proteins); 0 (Staphylococcal Protein A); EC 3.4.21.64 (Endopeptidase K)
[Em] Mês de entrada:1801
[Cu] Atualização por classe:180130
[Lr] Data última revisão:
180130
[Sb] Subgrupo de revista:IM
[Da] Data de entrada para processamento:171208
[St] Status:MEDLINE


  2 / 4372 MEDLINE  
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[PMID]:28859130
[Au] Autor:Balachandran M; Giannone RJ; Bemis DA; Kania SA
[Ad] Endereço:Department of Biomedical and Diagnostic Sciences, The University of Tennessee, Knoxville, Tennessee, United States of America.
[Ti] Título:Molecular basis of surface anchored protein A deficiency in the Staphylococcus aureus strain Wood 46.
[So] Source:PLoS One;12(8):e0183913, 2017.
[Is] ISSN:1932-6203
[Cp] País de publicação:United States
[La] Idioma:eng
[Ab] Resumo:Protein A in Staphylococcus aureus is encoded by the spa (staphylococcal protein A) gene and binds to immunoglobulin (Ig). The S. aureus strain Wood 46 has been variously reported as protein A-deficient and/or spa negative and used as a control in animal models of staphylococcal infections. The results of this study indicate that Wood 46 has normal spa expression but transcribes very low levels of the srtA gene which encodes the sortase A (SrtA) enzyme. This is consistent with unique mutations in the srtA promoter. In this study, a low level of sortase A explains deficient anchoring of proteins with an LPXTG motif, such as protein A, fibrinogen-binding protein and fibronectin-binding proteins A and B on to the peptidoglycan cell wall. The activity of secreted protein A is an important consideration for use of Wood 46 in functional experiments and animal models.
[Mh] Termos MeSH primário: Adesinas Bacterianas/genética
Aminoaciltransferases/genética
Proteínas de Bactérias/genética
Proteínas de Transporte/genética
Cisteína Endopeptidases/genética
Regulação Bacteriana da Expressão Gênica
Proteína Estafilocócica A/genética
Staphylococcus aureus/genética
[Mh] Termos MeSH secundário: Adesinas Bacterianas/metabolismo
Motivos de Aminoácidos
Aminoaciltransferases/metabolismo
Proteínas de Bactérias/metabolismo
Sítios de Ligação
Proteínas de Transporte/metabolismo
Parede Celular/química
Parede Celular/metabolismo
Cisteína Endopeptidases/metabolismo
Mutação
Regiões Promotoras Genéticas
Ligação Proteica
Proteína Estafilocócica A/metabolismo
Staphylococcus aureus/metabolismo
Transcrição Genética
[Pt] Tipo de publicação:JOURNAL ARTICLE
[Nm] Nome de substância:
0 (Adhesins, Bacterial); 0 (Bacterial Proteins); 0 (Carrier Proteins); 0 (Staphylococcal Protein A); 0 (fibrinogen-binding protein A, Staphylococcus aureus); 0 (fibronectin-binding proteins, bacterial); EC 2.3.2.- (Aminoacyltransferases); EC 2.3.2.- (sortase A); EC 3.4.22.- (Cysteine Endopeptidases)
[Em] Mês de entrada:1710
[Cu] Atualização por classe:171016
[Lr] Data última revisão:
171016
[Sb] Subgrupo de revista:IM
[Da] Data de entrada para processamento:170901
[St] Status:MEDLINE
[do] DOI:10.1371/journal.pone.0183913


  3 / 4372 MEDLINE  
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[PMID]:28690180
[Au] Autor:Wang J; Wang J; Zhang Z; Zhang X; Ru S; Dong Y
[Ad] Endereço:Marine Life Science College, Ocean University of China, Qingdao 266003, China.
[Ti] Título:Development of an immunosensor for quantifying zebrafish vitellogenin based on the Octet system.
[So] Source:Anal Biochem;533:60-65, 2017 Sep 15.
[Is] ISSN:1096-0309
[Cp] País de publicação:United States
[La] Idioma:eng
[Ab] Resumo:Vitellogenin (Vtg) is a sensitive biomarker for environmental estrogens. In this study, an immunosensor for quantifying zebrafish Vtg was developed using the Octet system. First, Protein A sensors were immobilized with purified anti-lipovitellin (Lv) antibody that demonstrated specificity to Vtg. Then, antibody-coated biosensors were immersed into zebrafish Lv standards and diluted samples. The Octet system measured and recorded kinetic parameters between antigens and captured antibody within 5 min. Sample Vtg concentrations were automatically calculated by interpolating relative binding rates observed with each sample and the immobilized anti-Lv antibody into the developed standard curve. The sensor arrays exhibited a wide linear range from 78 to 5000 ng/mL, and the inter-assay coefficient of variation was 0.66-1.97%. Furthermore, the performance of the immunosensor in detecting Vtg was evaluated by quantifying Vtg induction in juvenile zebrafish exposed to 17ß-estradiol (E ). Compared with conventional immunoassay techniques, the Vtg immunosensor developed based on the Octet system was much simpler and less time-consuming, allowing rapid Vtg quantification within 15 min. Moreover, Protein A sensors could be reused many times to ensure that the assays have high reproducibility. Therefore, we suggest that immunosensors based on the Octet system are an easily operated detection method for ecotoxicological research.
[Mh] Termos MeSH primário: Anticorpos Anti-Idiotípicos/isolamento & purificação
Técnicas Biossensoriais
Vitelogeninas/isolamento & purificação
[Mh] Termos MeSH secundário: Animais
Anticorpos Anti-Idiotípicos/imunologia
Proteínas do Ovo/imunologia
Estradiol/farmacologia
Proteína Estafilocócica A/química
Vitelogeninas/imunologia
Peixe-Zebra/imunologia
[Pt] Tipo de publicação:JOURNAL ARTICLE
[Nm] Nome de substância:
0 (Antibodies, Anti-Idiotypic); 0 (Egg Proteins); 0 (Staphylococcal Protein A); 0 (Vitellogenins); 4TI98Z838E (Estradiol); 9088-43-1 (lipovitellin)
[Em] Mês de entrada:1708
[Cu] Atualização por classe:170804
[Lr] Data última revisão:
170804
[Sb] Subgrupo de revista:IM
[Da] Data de entrada para processamento:170711
[St] Status:MEDLINE


  4 / 4372 MEDLINE  
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[PMID]:28674338
[Au] Autor:Mahara A; Harada-Shiba M; Yamaoka T
[Ad] Endereço:Department of Biomedical Engineering, National Cerebral and Cardiovascular Center Research Institute.
[Ti] Título:A Novel Strategy for Etiologic Factor Removal: Drug-Navigated Clearance System (DNCS).
[So] Source:Chem Pharm Bull (Tokyo);65(7):649-652, 2017.
[Is] ISSN:1347-5223
[Cp] País de publicação:Japan
[La] Idioma:eng
[Ab] Resumo:Here, we propose a novel therapeutic concept named drug-navigated clearance system (DNCS), in which a "navigator" decreases the concentration of a target etiologic factor in the blood by steering it to an unusual metabolic pathway. The navigator is composed of protein A (ProA) and dextran sulfate (DexS) and it successfully navigated antibodies (ABs), a model etiologic factor of dilated cardiomyopathy, to hepatocytes in vitro in the presence of low-density lipoprotein (LDL). ProA captured the Fc region of the target antibody while the DexS bound to LDL via the well-known electrostatic interaction. The hepatocytes simultaneously took up LDL via the LDL-receptor and internalized the AB/ProA-DexS complex that was bound to LDL. Therefore, this process demonstrates our attempt to navigate the etiologic factor to an alternate target pathway such as the LDL salvage.
[Mh] Termos MeSH primário: Sulfato de Dextrana/metabolismo
Receptores de LDL/metabolismo
Proteína Estafilocócica A/metabolismo
[Mh] Termos MeSH secundário: Sulfato de Dextrana/química
Células Hep G2
Hepatócitos/metabolismo
Seres Humanos
Lipoproteínas LDL/metabolismo
Proteína Estafilocócica A/química
[Pt] Tipo de publicação:JOURNAL ARTICLE
[Nm] Nome de substância:
0 (LDLR protein, human); 0 (Lipoproteins, LDL); 0 (Receptors, LDL); 0 (Staphylococcal Protein A); 9042-14-2 (Dextran Sulfate)
[Em] Mês de entrada:1708
[Cu] Atualização por classe:170814
[Lr] Data última revisão:
170814
[Sb] Subgrupo de revista:IM
[Da] Data de entrada para processamento:170705
[St] Status:MEDLINE
[do] DOI:10.1248/cpb.c17-00282


  5 / 4372 MEDLINE  
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[PMID]:28668497
[Au] Autor:Inouye S; Sahara-Miura Y
[Ad] Endereço:Yokohama Research Center, JNC Co., 5-1 Okawa, Kanazawa-ku, Yokohama 236-8605, Japan. Electronic address: sinouye@jnc-corp.co.jp.
[Ti] Título:A fusion protein of the synthetic IgG-binding domain and aequorin: Expression and purification from E. coli cells and its application.
[So] Source:Protein Expr Purif;137:58-63, 2017 Sep.
[Is] ISSN:1096-0279
[Cp] País de publicação:United States
[La] Idioma:eng
[Ab] Resumo:Aequorin is a Ca -binding photoprotein that is a complex of apoaequorin (apoAQ) and 2-peroxycoelenterazine. In this study, the fusion protein (ZZ-apoAQ) composed of the synthetic IgG-binding domain (ZZ domain) derived from Staphylococcus aureus protein A and apoAQ was expressed into the periplasmic space of Escherichia coli cells. ZZ-apoAQ was highly purified using Ni-chelate affinity chromatography followed by IgG affinity chromatography. ZZ-AQ was prepared from purified ZZ-apoAQ by incubation with coelenterazine and was characterized, including its luminescence properties. ZZ-AQ could be used as a reporter for detecting IgG and the measurable range of IgG coated on a 96-well plate was 1-1000 ng/mL.
[Mh] Termos MeSH primário: Aquaporinas
Bioensaio/métodos
Expressão Gênica
Imunoglobulina G/análise
Proteínas Recombinantes de Fusão
Proteína Estafilocócica A
Staphylococcus aureus/genética
[Mh] Termos MeSH secundário: Aquaporinas/biossíntese
Aquaporinas/genética
Escherichia coli/genética
Escherichia coli/metabolismo
Seres Humanos
Proteínas Recombinantes de Fusão/biossíntese
Proteínas Recombinantes de Fusão/química
Proteínas Recombinantes de Fusão/genética
Proteína Estafilocócica A/biossíntese
Proteína Estafilocócica A/química
Proteína Estafilocócica A/genética
Staphylococcus aureus/metabolismo
[Pt] Tipo de publicação:JOURNAL ARTICLE
[Nm] Nome de substância:
0 (Aquaporins); 0 (Immunoglobulin G); 0 (Recombinant Fusion Proteins); 0 (Staphylococcal Protein A)
[Em] Mês de entrada:1708
[Cu] Atualização por classe:170807
[Lr] Data última revisão:
170807
[Sb] Subgrupo de revista:IM
[Da] Data de entrada para processamento:170703
[St] Status:MEDLINE


  6 / 4372 MEDLINE  
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[PMID]:28575112
[Au] Autor:Park KH; Greenwood-Quaintance KE; Uhl JR; Cunningham SA; Chia N; Jeraldo PR; Sampathkumar P; Nelson H; Patel R
[Ad] Endereço:Division of Clinical Microbiology, Department of Laboratory Medicine and Pathology, Mayo Clinic, Rochester, Minnesota, United States of America.
[Ti] Título:Molecular epidemiology of Staphylococcus aureus bacteremia in a single large Minnesota medical center in 2015 as assessed using MLST, core genome MLST and spa typing.
[So] Source:PLoS One;12(6):e0179003, 2017.
[Is] ISSN:1932-6203
[Cp] País de publicação:United States
[La] Idioma:eng
[Ab] Resumo:Staphylococcus aureus is a leading cause of bacteremia in hospitalized patients. Whether or not S. aureus bacteremia (SAB) is associated with clonality, implicating potential nosocomial transmission, has not, however, been investigated. Herein, we examined the epidemiology of SAB using whole genome sequencing (WGS). 152 SAB isolates collected over the course of 2015 at a single large Minnesota medical center were studied. Staphylococcus protein A (spa) typing was performed by PCR/Sanger sequencing; multilocus sequence typing (MLST) and core genome MLST (cgMLST) were determined by WGS. Forty-eight isolates (32%) were methicillin-resistant S. aureus (MRSA). The isolates encompassed 66 spa types, clustered into 11 spa clonal complexes (CCs) and 10 singleton types. 88% of 48 MRSA isolates belonged to spa CC-002 or -008. Methicillin-susceptible S. aureus (MSSA) isolates were more genotypically diverse, with 61% distributed across four spa CCs (CC-002, CC-012, CC-008 and CC-084). By MLST, there was 31 sequence types (STs), including 18 divided into 6 CCs and 13 singleton STs. Amongst MSSA isolates, the common MLST clones were CC5 (23%), CC30 (19%), CC8 (15%) and CC15 (11%). Common MRSA clones were CC5 (67%) and CC8 (25%); there were no MRSA isolates in CC45 or CC30. By cgMLST analysis, there were 9 allelic differences between two isolates, with the remaining 150 isolates differing from each other by over 40 alleles. The two isolates were retroactively epidemiologically linked by medical record review. Overall, cgMLST analysis resulted in higher resolution epidemiological typing than did multilocus sequence or spa typing.
[Mh] Termos MeSH primário: Bacteriemia/diagnóstico
Bacteriemia/microbiologia
Infecções Estafilocócicas/diagnóstico
Infecções Estafilocócicas/microbiologia
Proteína Estafilocócica A/genética
Staphylococcus aureus/genética
[Mh] Termos MeSH secundário: Idoso
Antibacterianos/farmacologia
Bacteriemia/tratamento farmacológico
Bacteriemia/epidemiologia
Feminino
Genoma Bacteriano
Seres Humanos
Masculino
Staphylococcus aureus Resistente à Meticilina/efeitos dos fármacos
Staphylococcus aureus Resistente à Meticilina/genética
Staphylococcus aureus Resistente à Meticilina/isolamento & purificação
Meia-Idade
Minnesota/epidemiologia
Epidemiologia Molecular
Tipagem de Sequências Multilocus
Filogenia
Infecções Estafilocócicas/tratamento farmacológico
Infecções Estafilocócicas/epidemiologia
Staphylococcus aureus/efeitos dos fármacos
Staphylococcus aureus/isolamento & purificação
[Pt] Tipo de publicação:JOURNAL ARTICLE
[Nm] Nome de substância:
0 (Anti-Bacterial Agents); 0 (Staphylococcal Protein A)
[Em] Mês de entrada:1709
[Cu] Atualização por classe:170914
[Lr] Data última revisão:
170914
[Sb] Subgrupo de revista:IM
[Da] Data de entrada para processamento:170603
[St] Status:MEDLINE
[do] DOI:10.1371/journal.pone.0179003


  7 / 4372 MEDLINE  
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[PMID]:28502296
[Au] Autor:Jia L; Cao X; Yang Z
[Ad] Endereço:Departent of Pathogen Biology and Immunology, School of Basic Medical Sciences, Ningxia Medical University, Yinchuan 750004, China.
[Ti] Título:[Recombinant Legionella pneumophila flagella protein A (rflaA) induces the secretion of IL-6 and IL-1ß in RAW264.7 cells in vitro].
[So] Source:Xi Bao Yu Fen Zi Mian Yi Xue Za Zhi;33(5):601-605, 2017 May.
[Is] ISSN:1007-8738
[Cp] País de publicação:China
[La] Idioma:chi
[Ab] Resumo:Objective To investigate the effect of recombinant Legionella pneumophila flagella protein A (rflaA) on the secretion of interleukin-6 (IL-6) and interleukin-1ß (IL-1ß) by RAW264.7 macrophage and the possible mechanism. Methods RAW264.7 cells were treated with 0.000, 0.125, 0.250, 0.500, 1.000, 2.000, 4.000 and 8.000 µg/mL rflaA to determine the EC of rflaA using CCK-8 assay. Secretion of IL-6 and IL-1ß were measured by ELISA at 24, 36 and 48 hours after treatment of the cells with 0.04, 0.08 and 0.16 µg/mL rflaA. At 6, 12, 24, 36 and 48 hours after treatment of the cells with 0.04, 0.08 and 0.16 µg/mL rflaA, the expressions of IL-6, IL-1ß, NOD-like receptor protein 3 (NLRP3) and caspase-1 mRNAs were detected by quantitative real-time PCR, and the expressions of NLRP3 and caspase-1 proteins were tested by Western blotting. Results RflaA enhanced the expressions of IL-6 and IL-1ß, and the higher concentration of rflaA was more potential. The expressions of IL-6 and IL-1ß reached peak when the cells were treated with 0.16 µg/mL rflaA for 36 hours. Treatment of RAW264.7 cells with rflaA promoted the expressions of IL-6 and IL-1ß, NLRP3 and caspase-1 mRNA, and 0.16 µg/mL rflaA was the most potential at 12 hours after treatment. Expressions of NLRP3 and caspase-1 protein increased after treatment with rflaA, and 0.16 µg/mL rflaA induced the highest expression of both proteins at 24 hours after treatment. Conclusion RflaA could enhance the secretion of IL-6 and IL-1ß by promoting the expressions of NLRP3 and caspase-1 in RAW264.7 cells.
[Mh] Termos MeSH primário: Flagelos/metabolismo
Interleucina-1beta/metabolismo
Interleucina-6/metabolismo
Legionella pneumophila/metabolismo
Proteínas Recombinantes/farmacologia
Proteína Estafilocócica A/farmacologia
[Mh] Termos MeSH secundário: Animais
Linhagem Celular
Legionella pneumophila/genética
Camundongos
Células RAW 264.7
Proteínas Recombinantes/genética
Proteína Estafilocócica A/genética
[Pt] Tipo de publicação:JOURNAL ARTICLE
[Nm] Nome de substância:
0 (Interleukin-1beta); 0 (Interleukin-6); 0 (Recombinant Proteins); 0 (Staphylococcal Protein A)
[Em] Mês de entrada:1710
[Cu] Atualização por classe:171004
[Lr] Data última revisão:
171004
[Sb] Subgrupo de revista:IM
[Da] Data de entrada para processamento:170516
[St] Status:MEDLINE


  8 / 4372 MEDLINE  
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[PMID]:28494950
[Au] Autor:Rahamim G; Amir D; Haas E
[Ad] Endereço:The Goodman Faculty of Life Sciences Bar Ilan University, Ramat Gan, Israel.
[Ti] Título:Simultaneous Determination of Two Subdomain Folding Rates Using the "Transfer-Quench" Method.
[So] Source:Biophys J;112(9):1786-1796, 2017 May 09.
[Is] ISSN:1542-0086
[Cp] País de publicação:United States
[La] Idioma:eng
[Ab] Resumo:The investigation of the mechanism of protein folding is complicated by the context dependence of the rates of intramolecular contact formation. Methods based on site-specific labeling and ultrafast spectroscopic detection of fluorescence signals were developed for monitoring the rates of individual subdomain folding transitions in situ, in the context of the whole molecule. However, each site-specific labeling modification might affect rates of folding of near-neighbor structural elements, and thus limit the ability to resolve fine differences in rates of folding of these elements. Therefore, it is highly desirable to be able to study the rates of folding of two or more neighboring subdomain structures using a single mutant to facilitate resolution of the order and interdependence of such steps. Here, we report the development of the "Transfer-Quench" method for measuring the rate of formation of two structural elements using a single triple-labeled mutant. This method is based on Förster resonance energy transfer combined with fluorescence quenching. We placed the donor and acceptor at the loop ends, and a quencher at an α-helical element involved in the node forming the loop. The folding of the triple-labeled mutant is monitored by the acceptor emission. The formation of nonlocal contact (loop closure) increases the time-dependent acceptor emission, while the closure of the labeled helix turn reduces this emission. The method was applied in a study of the folding mechanism of the common model protein, the B domain of staphylococcal protein A. Only natural amino acids were used as probes, and thus possible structural perturbations were minimized. Tyr and Trp residues served as donor and acceptor at the ends of a long loop between helices I and II, and a Cys residue as a quencher for the acceptor. We found that the closure of the loop (segment 14-33) occurs with the same rate constant as the nucleation of helix HII (segment 33-29), in line with the nucleation-condensation model.
[Mh] Termos MeSH primário: Imagem Molecular/métodos
Domínios Proteicos
Dobramento de Proteína
Proteína Estafilocócica A/química
[Mh] Termos MeSH secundário: Algoritmos
Escherichia coli
Transferência Ressonante de Energia de Fluorescência
Cinética
Mutação
Domínios Proteicos/genética
Proteína Estafilocócica A/genética
Staphylococcus
[Pt] Tipo de publicação:JOURNAL ARTICLE
[Nm] Nome de substância:
0 (Staphylococcal Protein A)
[Em] Mês de entrada:1708
[Cu] Atualização por classe:170808
[Lr] Data última revisão:
170808
[Sb] Subgrupo de revista:IM
[Da] Data de entrada para processamento:170513
[St] Status:MEDLINE


  9 / 4372 MEDLINE  
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[PMID]:28437636
[Au] Autor:Zhou Y; Kearney CM
[Ad] Endereço:Institute of Biomedical Studies, Baylor University, Waco, TX, USA. Electronic address: Yiyang_Zhou@baylor.edu.
[Ti] Título:Chimeric Flock House virus protein A with endoplasmic reticulum-targeting domain enhances viral replication and virus-like particle trans-encapsidation in plants.
[So] Source:Virology;507:151-160, 2017 Jul.
[Is] ISSN:1096-0341
[Cp] País de publicação:United States
[La] Idioma:eng
[Ab] Resumo:Flock House virus (FHV) RNA can be trans-encapsidated, entirely in planta, by tobacco mosaic virus coat protein to form virus-like particles (VLPs). Vaccination with these VLPs leads to strong antigen expression in mice and immune-activation. We hypothesize that creating an additional cellular site for replication and/or trans-encapsidation might significantly improve the final output of trans-encapsidated product. FHV protein A was engineered to target the endoplasmic reticulum (ER) via a heterologous tobacco etch virus ER-targeting domain, and was expressed in cis or in trans relative to the replicating FHV RNA1. A strong increase in marker gene expression in plants was noted when ER-targeted protein A was supplied in trans. RNA fluorescence in situ hybridization revealed RNA1 replication in both the mitochondria and ER, and total RNA1 accumulation was increased. In support of our hypothesis, VLP yield was increased significantly by the addition of this single genetic component to the inoculum.
[Mh] Termos MeSH primário: Retículo Endoplasmático/virologia
Nodaviridae/fisiologia
Proteína Estafilocócica A/química
Proteína Estafilocócica A/metabolismo
Tabaco/virologia
Proteínas Virais/química
Proteínas Virais/metabolismo
Replicação Viral
[Mh] Termos MeSH secundário: Mitocôndrias/virologia
Nodaviridae/química
Nodaviridae/genética
Doenças das Plantas/virologia
Domínios Proteicos
Transporte Proteico
RNA Viral/genética
RNA Viral/metabolismo
Proteína Estafilocócica A/genética
Proteínas Virais/genética
[Pt] Tipo de publicação:JOURNAL ARTICLE
[Nm] Nome de substância:
0 (RNA, Viral); 0 (Staphylococcal Protein A); 0 (Viral Proteins)
[Em] Mês de entrada:1707
[Cu] Atualização por classe:170717
[Lr] Data última revisão:
170717
[Sb] Subgrupo de revista:IM
[Da] Data de entrada para processamento:170425
[St] Status:MEDLINE


  10 / 4372 MEDLINE  
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[PMID]:28410804
[Au] Autor:Mazzer AR; Clifton LA; Perevozchikova T; Butler PD; Roberts CJ; Bracewell DG
[Ad] Endereço:Dept. Biochemical Engineering, University College London, Gower Street, London, WC1E 6BT, UK.
[Ti] Título:Neutron reflectivity measurement of protein A-antibody complex at the solid-liquid interface.
[So] Source:J Chromatogr A;1499:118-131, 2017 May 26.
[Is] ISSN:1873-3778
[Cp] País de publicação:Netherlands
[La] Idioma:eng
[Ab] Resumo:Chromatography is a ubiquitous unit operation in the purification of biopharmaceuticals yet few studies have addressed the biophysical characterisation of proteins at the solution-resin interface. Chromatography and other adsorption and desorption processes have been shown to induce protein aggregation which is undesirable in biopharmaceutical products. In order to advance understanding of how adsorption processes might impact protein stability, neutron reflectivity was used to characterise the structure of adsorbed immunoglobulin G (IgG) on model surfaces. In the first model system, IgG was adsorbed directly to silica and demonstrated a side-on orientation with high surface contact. A maximum dimension of 60Å in the surface normal direction and high density surface coverage were observed under pH 4.1 conditions. In chromatography buffers, pH was found to influence IgG packing density and orientation at the solid-liquid interface. In the second model system, which was designed to mimic an affinity chromatography surface, protein A was attached to a silica surface to produce a configuration representative of a porous glass chromatography resin. Interfacial structure was probed during sequential stages from ligand attachment, through to IgG binding and elution. Adsorbed IgG structures extended up to 250Å away from the surface and showed dependence on surface blocking strategies. The data was suggestive of two IgG molecules bound to protein A with a somewhat skewed orientation and close proximity to the silica surface. The findings provide insight into the orientation of adsorbed antibody structures under conditions encountered during chromatographic separations.
[Mh] Termos MeSH primário: Proteína Estafilocócica A/química
[Mh] Termos MeSH secundário: Adsorção
Cromatografia Líquida/instrumentação
Imunoglobulina G/química
Nêutrons
Ligação Proteica
Estabilidade Proteica
Dióxido de Silício/química
Propriedades de Superfície
[Pt] Tipo de publicação:JOURNAL ARTICLE
[Nm] Nome de substância:
0 (Immunoglobulin G); 0 (Staphylococcal Protein A); 7631-86-9 (Silicon Dioxide)
[Em] Mês de entrada:1710
[Cu] Atualização por classe:171016
[Lr] Data última revisão:
171016
[Sb] Subgrupo de revista:IM
[Da] Data de entrada para processamento:170416
[St] Status:MEDLINE



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