Base de dados : MEDLINE
Pesquisa : D01.248.497.158.874.390 [Categoria DeCS]
Referências encontradas : 16628 [refinar]
Mostrando: 1 .. 10   no formato [Detalhado]

página 1 de 1663 ir para página                         

  1 / 16628 MEDLINE  
              next record last record
seleciona
para imprimir
Fotocópia
Texto completo
[PMID]:29367490
[Au] Autor:Asakawa T; Takano Y; Ohta A; Asakawa H
[Ad] Endereço:School of Chemistry, College of Science and Engineering, Kanazawa University.
[Ti] Título:Aggregation and pH Responsive Behavior of Thioester Surfactants and Formation of Disulfide Linkages in Aqueous Solutions.
[So] Source:J Oleo Sci;67(2):199-206, 2018 Feb 01.
[Is] ISSN:1347-3352
[Cp] País de publicação:Japan
[La] Idioma:eng
[Ab] Resumo:pH responsive surfactants, [C H N(CH ) (CH ) SCOCH ]Br (C nSAc, n = 4, 11, 12), were prepared, and their properties in aqueous solution were examined. The critical micelle concentration (cmc) and critical vesicle concentration (cvc) were determined based on changes in conductivity, as well as by fluorescence measurements, and light scattering methods. A significant increase in the light scattering intensities of the C nSAc (n=11, 12) systems suggested that the growth of aggregates was accompanied by considerable counterion binding with increasing surfactant concentration. The diameter of C 11SAc, recorded by the dynamic light scattering measurements, was about 9.6 ±1.0 nm, which was slightly smaller than that for didodecyldimethylammonium bromide (DDAB) vesicles. The thioester group was easily hydrolyzed upon the addition of NaOH, while it was hardly hydrolyzed with the addition of HCl. The time course of alkaline hydrolysis was examined by the conductivity measurements and high-performance liquid chromatography analysis. [C H N(CH ) (CH ) SS(CH ) N(CH ) C H ]2Br (2C 11SS) was generated in the C 11SAc alkaline solution because of air oxidation. The C 11SAc alkaline solution gradually became an opaque blue color with increasing light scattering at 346 nm, indicating the remarkable growth of vesicles. The chemical structure of 2C 11SS was consistent with that of a disulfide linked double tailed surfactant, similar to DDAB. The disulfide linkage between the double tailed surfactants will contribute to the stabilization and growth of vesicles.
[Mh] Termos MeSH primário: Ésteres/química
Tensoativos/química
Água/química
[Mh] Termos MeSH secundário: Dissulfetos/química
Difusão Dinâmica da Luz
Concentração de Íons de Hidrogênio
Hidrólise
Micelas
Oxirredução
Compostos de Amônio Quaternário/química
Soluções
Tensão Superficial
[Pt] Tipo de publicação:JOURNAL ARTICLE
[Nm] Nome de substância:
0 (Disulfides); 0 (Esters); 0 (Micelles); 0 (Quaternary Ammonium Compounds); 0 (Solutions); 0 (Surface-Active Agents); 059QF0KO0R (Water); 13146-86-6 (didodecyldimethylammonium)
[Em] Mês de entrada:1803
[Cu] Atualização por classe:180309
[Lr] Data última revisão:
180309
[Sb] Subgrupo de revista:IM
[Da] Data de entrada para processamento:180126
[St] Status:MEDLINE
[do] DOI:10.5650/jos.ess17192


  2 / 16628 MEDLINE  
              first record previous record next record last record
seleciona
para imprimir
Fotocópia
Texto completo
[PMID]:29216817
[Au] Autor:Nivala O; Faccio G; Arvas M; Permi P; Buchert J; Kruus K; Mattinen ML
[Ad] Endereço:VTT Technical Research Centre of Finland, Ltd., P.O. Box 1000, FI-02044, Espoo, Finland. outi.nivala@helsinki.fi.
[Ti] Título:Characterization of sulfhydryl oxidase from Aspergillus tubingensis.
[So] Source:BMC Biochem;18(1):15, 2017 12 08.
[Is] ISSN:1471-2091
[Cp] País de publicação:England
[La] Idioma:eng
[Ab] Resumo:BACKGROUND: Despite of the presence of sulfhydryl oxidases (SOXs) in the secretomes of industrially relevant organisms and their many potential applications, only few of these enzymes have been biochemically characterized. In addition, basic functions of most of the SOX enzymes reported so far are not fully understood. In particular, the physiological role of secreted fungal SOXs is unclear. RESULTS: The recently identified SOX from Aspergillus tubingensis (AtSOX) was produced, purified and characterized in the present work. AtSOX had a pH optimum of 6.5, and showed a good pH stability retaining more than 80% of the initial activity in a pH range 4-8.5 within 20 h. More than 70% of the initial activity was retained after incubation at 50 °C for 20 h. AtSOX contains a non-covalently bound flavin cofactor. The enzyme oxidised a sulfhydryl group of glutathione to form a disulfide bond, as verified by nuclear magnetic resonance spectroscopy. AtSOX preferred glutathione as a substrate over cysteine and dithiothreitol. The activity of the enzyme was totally inhibited by 10 mM zinc sulphate. Peptide- and protein-bound sulfhydryl groups in bikunin, gliotoxin, holomycin, insulin B chain, and ribonuclease A, were not oxidised by the enzyme. Based on the analysis of 33 fungal genomes, SOX enzyme encoding genes were found close to nonribosomal peptide synthetases (NRPS) but not with polyketide synthases (PKS). In the phylogenetic tree, constructed from 25 SOX and thioredoxin reductase sequences from IPR000103 InterPro family, AtSOX was evolutionary closely related to other Aspergillus SOXs. Oxidoreductases involved in the maturation of nonribosomal peptides of fungal and bacterial origin, namely GliT, HlmI and DepH, were also evolutionary closely related to AtSOX whereas fungal thioreductases were more distant. CONCLUSIONS: AtSOX (55 kDa) is a fungal secreted flavin-dependent enzyme with good stability to both pH and temperature. A Michaelis-Menten behaviour was observed with reduced glutathione as a substrate. Based on the location of SOX enzyme encoding genes close to NRPSs, SOXs could be involved in the secondary metabolism and act as an accessory enzyme in the production of nonribosomal peptides.
[Mh] Termos MeSH primário: Aspergillus/enzimologia
Oxirredutases/metabolismo
[Mh] Termos MeSH secundário: Dissulfetos
Estabilidade Enzimática
Glutationa/metabolismo
Concentração de Íons de Hidrogênio
Oxirredutases/química
Oxirredutases/genética
Oxirredutases/isolamento & purificação
Peptídeo Sintases
Especificidade por Substrato
[Pt] Tipo de publicação:JOURNAL ARTICLE; RESEARCH SUPPORT, NON-U.S. GOV'T
[Nm] Nome de substância:
0 (Disulfides); EC 1.- (Oxidoreductases); EC 1.8.3.- (sulfhydryl oxidase); EC 6.3.2.- (Peptide Synthases); EC 6.3.2.- (non-ribosomal peptide synthase); GAN16C9B8O (Glutathione)
[Em] Mês de entrada:1712
[Cu] Atualização por classe:180309
[Lr] Data última revisão:
180309
[Sb] Subgrupo de revista:IM
[Da] Data de entrada para processamento:171209
[St] Status:MEDLINE
[do] DOI:10.1186/s12858-017-0090-4


  3 / 16628 MEDLINE  
              first record previous record next record last record
seleciona
para imprimir
Fotocópia
Texto completo
[PMID]:28746421
[Au] Autor:Sadowska-Bartosz I; Furmaniak P; Bieszczad-Bedrejczuk E; Bartosz G; Glowacki R
[Ad] Endereço:Department of Analytical Biochemistry, Faculty of Biology and Agriculture, University of Rzeszow, Rzeszów, Poland.
[Ti] Título:Developmental changes in the levels and redox potentials of main hemolymph thiols/disulfides in the Jamaican field cricket Gryllus assimilis.
[So] Source:Acta Biochim Pol;64(3):503-506, 2017.
[Is] ISSN:1734-154X
[Cp] País de publicação:Poland
[La] Idioma:eng
[Ab] Resumo:Main thiols and disulfides were determined in the hemolymph of the Jamaican field cricket Gryllus assimilis at various developmental stages. On the basis of these data, redox potentials of the glutathione, cysteine and homocysteine redox systems were calculated. The concentrations of all thiols studied decreased during development (at a stage of 6 molts) with respect to young crickets, and increased again in adult insects. Redox potentials of the glutathione and cysteine systems increased from values of -131.0±5.6 mV and -86.9±17.1 mV, respectively in young crickets to -58.0±3.6 mV and -36.1±4.2 mV, respectively, at the stage of 6 molts and decreased to values of -110.4±24.8 mV and -66.3±12.2 mV, respectively, in adult insects. Redox potentials of the glutathione and cysteine systems in the hemolymph of young and adult insects were similar to those reported for human plasma.
[Mh] Termos MeSH primário: Dissulfetos/metabolismo
Gryllidae/crescimento & desenvolvimento
Gryllidae/metabolismo
Hemolinfa/metabolismo
Compostos de Sulfidrila/metabolismo
[Mh] Termos MeSH secundário: Animais
Cisteína/metabolismo
Glutationa/metabolismo
Dissulfeto de Glutationa/metabolismo
Homocisteína/metabolismo
Ninfa/crescimento & desenvolvimento
Ninfa/metabolismo
Oxirredução
[Pt] Tipo de publicação:JOURNAL ARTICLE
[Nm] Nome de substância:
0 (Disulfides); 0 (Sulfhydryl Compounds); 0LVT1QZ0BA (Homocysteine); GAN16C9B8O (Glutathione); K848JZ4886 (Cysteine); ULW86O013H (Glutathione Disulfide)
[Em] Mês de entrada:1802
[Cu] Atualização por classe:180228
[Lr] Data última revisão:
180228
[Sb] Subgrupo de revista:IM
[Da] Data de entrada para processamento:170727
[St] Status:MEDLINE
[do] DOI:10.18388/abp.2017_1510


  4 / 16628 MEDLINE  
              first record previous record next record last record
seleciona
para imprimir
Fotocópia
Texto completo
[PMID]:28468269
[Au] Autor:Logashina YA; Solstad RG; Mineev KS; Korolkova YV; Mosharova IV; Dyachenko IA; Palikov VA; Palikova YA; Murashev AN; Arseniev AS; Kozlov SA; Stensvåg K; Haug T; Andreev YA
[Ad] Endereço:Shemyakin-Ovchinnikov Institute of Bioorganic Chemistry, Russian Academy of Sciences, ul. Miklukho-Maklaya 16/10, 117997 Moscow, Russia. yulia.logashina@gmail.com.
[Ti] Título:New Disulfide-Stabilized Fold Provides Sea Anemone Peptide to Exhibit Both Antimicrobial and TRPA1 Potentiating Properties.
[So] Source:Toxins (Basel);9(5), 2017 Apr 29.
[Is] ISSN:2072-6651
[Cp] País de publicação:Switzerland
[La] Idioma:eng
[Ab] Resumo:A novel bioactive peptide named τ-AnmTx Ueq 12-1 (short name Ueq 12-1) was isolated and characterized from the sea anemone Ueq 12-1 is unique among the variety of known sea anemone peptides in terms of its primary and spatial structure. It consists of 45 amino acids including 10 cysteine residues with an unusual distribution and represents a new group of sea anemone peptides. The 3D structure of Ueq 12-1, determined by NMR spectroscopy, represents a new disulfide-stabilized fold partly similar to the defensin-like fold. Ueq 12-1 showed the dual activity of both a moderate antibacterial activity against Gram-positive bacteria and a potentiating activity on the transient receptor potential ankyrin 1 (TRPA1). Ueq 12-1 is a unique peptide potentiator of the TRPA1 receptor that produces analgesic and anti-inflammatory effects . The antinociceptive properties allow us to consider Ueq 12-1 as a potential analgesic drug lead with antibacterial properties.
[Mh] Termos MeSH primário: Analgésicos
Antibacterianos
Anti-Inflamatórios
Peptídeos
Anêmonas-do-Mar
Canal de Cátion TRPA1/metabolismo
[Mh] Termos MeSH secundário: Sequência de Aminoácidos
Analgésicos/química
Analgésicos/isolamento & purificação
Analgésicos/farmacologia
Analgésicos/uso terapêutico
Animais
Antibacterianos/química
Antibacterianos/isolamento & purificação
Antibacterianos/farmacologia
Antibacterianos/uso terapêutico
Anti-Inflamatórios/química
Anti-Inflamatórios/isolamento & purificação
Anti-Inflamatórios/farmacologia
Anti-Inflamatórios/uso terapêutico
Dissulfetos/química
Edema/tratamento farmacológico
Peptídeos/química
Peptídeos/isolamento & purificação
Peptídeos/farmacologia
Peptídeos/uso terapêutico
[Pt] Tipo de publicação:JOURNAL ARTICLE
[Nm] Nome de substância:
0 (Analgesics); 0 (Anti-Bacterial Agents); 0 (Anti-Inflammatory Agents); 0 (Disulfides); 0 (Peptides); 0 (TRPA1 Cation Channel)
[Em] Mês de entrada:1802
[Cu] Atualização por classe:180222
[Lr] Data última revisão:
180222
[Sb] Subgrupo de revista:IM
[Da] Data de entrada para processamento:170505
[St] Status:MEDLINE


  5 / 16628 MEDLINE  
              first record previous record next record last record
seleciona
para imprimir
Fotocópia
Texto completo
[PMID]:28470604
[Au] Autor:Saez NJ; Cristofori-Armstrong B; Anangi R; King GF
[Ad] Endereço:Institute for Molecular Bioscience, The University of Queensland, 306 Carmody Road, St. Lucia, QLD, 4067, Australia. n.saez@imb.uq.edu.au.
[Ti] Título:A Strategy for Production of Correctly Folded Disulfide-Rich Peptides in the Periplasm of E. coli.
[So] Source:Methods Mol Biol;1586:155-180, 2017.
[Is] ISSN:1940-6029
[Cp] País de publicação:United States
[La] Idioma:eng
[Ab] Resumo:Recombinant expression of disulfide-reticulated peptides and proteins is often challenging. We describe a method that exploits the periplasmic disulfide-bond forming machinery of Escherichia coli and combines this with a cleavable, solubility-enhancing fusion tag to obtain higher yields of correctly folded target protein than is achievable via cytoplasmic expression. The protocols provided herein cover all aspects of this approach, from vector construction and transformation to purification of the cleaved target protein and subsequent quality control.
[Mh] Termos MeSH primário: Dissulfetos/química
Escherichia coli/genética
Peptídeos/química
Peptídeos/genética
Periplasma/genética
[Mh] Termos MeSH secundário: Cromatografia de Afinidade/métodos
Cromatografia Líquida de Alta Pressão/métodos
Dissulfetos/isolamento & purificação
Dissulfetos/metabolismo
Eletroforese em Gel de Poliacrilamida/métodos
Peptídeos/isolamento & purificação
Plasmídeos/genética
Dobramento de Proteína
Proteínas Recombinantes de Fusão/química
Proteínas Recombinantes de Fusão/genética
Solubilidade
Transformação Genética
[Pt] Tipo de publicação:JOURNAL ARTICLE
[Nm] Nome de substância:
0 (Disulfides); 0 (Peptides); 0 (Recombinant Fusion Proteins)
[Em] Mês de entrada:1802
[Cu] Atualização por classe:180220
[Lr] Data última revisão:
180220
[Sb] Subgrupo de revista:IM
[Da] Data de entrada para processamento:170505
[St] Status:MEDLINE
[do] DOI:10.1007/978-1-4939-6887-9_10


  6 / 16628 MEDLINE  
              first record previous record next record last record
seleciona
para imprimir
Fotocópia
Texto completo
[PMID]:28470603
[Au] Autor:Besir H
[Ad] Endereço:Protein Expression and Purification Core Facility, EMBL Heidelberg, Meyerhofstraße 1, Heidelberg, 69117, Germany. besir@embl.de.
[Ti] Título:A Generic Protocol for Purifying Disulfide-Bonded Domains and Random Protein Fragments Using Fusion Proteins with SUMO3 and Cleavage by SenP2 Protease.
[So] Source:Methods Mol Biol;1586:141-154, 2017.
[Is] ISSN:1940-6029
[Cp] País de publicação:United States
[La] Idioma:eng
[Ab] Resumo:Recombinant expression of heterologous proteins in E. coli is well established for a wide range of proteins, although in many cases, purifying soluble and properly folded proteins remains challenging (Sorensen and Mortensen, J Biotechnol 115:113-128, 2005; Correa and Oppezzo, Methods Mol Biol 1258:27-44, 2015). Proteins that contain disulfide bonds (e.g., cytokines, growth factors) are often particularly difficult to purify in soluble form and still need optimizing of protocols in almost every step of the process (Berkmen, Protein Expr Purif 82:240-251, 2012; de Marco, Microb Cell Fact 11:129, 2012). Expression of disulfide bonded proteins in the periplasm of E. coli is one approach that can help to obtain soluble protein with the correct disulfide bridges forming in the periplasm. This offers the appropriate conditions for disulfide formation although periplasmic expression can also result in low expression levels and incorrect folding of the target protein (Schlapschy and Skerra, Methods Mol Biol 705:211-224, 2011). Generation of specific antibodies often requires a specific antigenic sequence of a protein in order to get an efficient immune response and minimize cross-reactivity of antibodies. Larger proteins like GST (Glutathione-S-transferase) or MBP (maltose binding protein) as solubilizing fusion partners are frequently used to keep antigens soluble and immunize animals. This approach has the disadvantage that the immune response against the fusion partner leads to additional antibodies that need to be separated from the antigen-specific antibodies. For both classes of proteins mentioned above, a protocol has been developed and optimized using the human version of small ubiquitin-like modifier 3 (SUMO3) protein and its corresponding protease SenP2. This chapter describes the experimental steps for expression, purification, refolding, and cleavage that are applicable to both disulfide-bonded proteins with a defined structure and random protein fragments for antibody generation or larger peptides with defined sequence that are difficult express on their own.
[Mh] Termos MeSH primário: Cisteína Endopeptidases/genética
Dissulfetos/química
Escherichia coli/genética
Ubiquitinas/genética
[Mh] Termos MeSH secundário: Sequência de Aminoácidos
Clonagem Molecular/métodos
Cisteína Endopeptidases/química
Cisteína Endopeptidases/metabolismo
Dissulfetos/metabolismo
Escherichia coli/metabolismo
Seres Humanos
Domínios Proteicos
Redobramento de Proteína
Proteínas Recombinantes de Fusão/química
Proteínas Recombinantes de Fusão/genética
Proteínas Recombinantes de Fusão/metabolismo
Solubilidade
Ubiquitinas/química
Ubiquitinas/metabolismo
[Pt] Tipo de publicação:JOURNAL ARTICLE
[Nm] Nome de substância:
0 (Disulfides); 0 (Recombinant Fusion Proteins); 0 (SUMO3 protein, human); 0 (Ubiquitins); EC 3.4.22.- (Cysteine Endopeptidases); EC 3.4.22.- (SENP2 protein, human)
[Em] Mês de entrada:1802
[Cu] Atualização por classe:180220
[Lr] Data última revisão:
180220
[Sb] Subgrupo de revista:IM
[Da] Data de entrada para processamento:170505
[St] Status:MEDLINE
[do] DOI:10.1007/978-1-4939-6887-9_9


  7 / 16628 MEDLINE  
              first record previous record next record last record
seleciona
para imprimir
Fotocópia
Texto completo
[PMID]:27778400
[Au] Autor:Kuan SL; Wang T; Weil T
[Ad] Endereço:Institute of Organic Chemistry III, Ulm University, Albert-Einstein-Allee 11, 89081, Ulm, Germany.
[Ti] Título:Site-Selective Disulfide Modification of Proteins: Expanding Diversity beyond the Proteome.
[So] Source:Chemistry;22(48):17112-17129, 2016 Nov 21.
[Is] ISSN:1521-3765
[Cp] País de publicação:Germany
[La] Idioma:eng
[Ab] Resumo:The synthetic transformation of polypeptides with molecular accuracy holds great promise for providing functional and structural diversity beyond the proteome. Consequently, the last decade has seen an exponential growth of site-directed chemistry to install additional features into peptides and proteins even inside living cells. The disulfide rebridging strategy has emerged as a powerful tool for site-selective modifications since most proteins contain disulfide bonds. In this Review, we present the chemical design, advantages and limitations of the disulfide rebridging reagents, while summarizing their relevance for synthetic customization of functional protein bioconjugates, as well as the resultant impact and advancement for biomedical applications.
[Mh] Termos MeSH primário: Dissulfetos/química
Peptídeos/química
Proteínas/química
[Mh] Termos MeSH secundário: Seres Humanos
Proteoma
[Pt] Tipo de publicação:JOURNAL ARTICLE; REVIEW
[Nm] Nome de substância:
0 (Disulfides); 0 (Peptides); 0 (Proteins); 0 (Proteome)
[Em] Mês de entrada:1802
[Cu] Atualização por classe:180220
[Lr] Data última revisão:
180220
[Sb] Subgrupo de revista:IM
[Da] Data de entrada para processamento:161026
[St] Status:MEDLINE
[do] DOI:10.1002/chem.201602298


  8 / 16628 MEDLINE  
              first record previous record next record last record
seleciona
para imprimir
Fotocópia
Texto completo
[PMID]:29208466
[Au] Autor:Dewangan J; Srivastava S; Mishra S; Pandey PK; Divakar A; Rath SK
[Ad] Endereço:Genotoxicity Lab, Division of Toxicology and Experimental Medicine, CSIR-Central Drug Research Institute, Lucknow, 226031, Uttar Pradesh, India. Electronic address: jayantdewangan@gmail.com.
[Ti] Título:Chetomin induces apoptosis in human triple-negative breast cancer cells by promoting calcium overload and mitochondrial dysfunction.
[So] Source:Biochem Biophys Res Commun;495(2):1915-1921, 2018 01 08.
[Is] ISSN:1090-2104
[Cp] País de publicação:United States
[La] Idioma:eng
[Ab] Resumo:Human triple-negative breast cancer (TNBC) is poorly diagnosed and unresponsive to conventional hormone therapy. Chetomin (CHET), a fungal metabolite synthesized by Chaetomium cochliodes, has been reported as a promising anticancer and antiangiogenic agent but the complete molecular mechanism of its anticancer potential remains to be elucidated. In our study, we explored the anti-neoplastic action of CHET on TNBC cells. Cytotoxicity studies were performed in human TNBC cells viz. MDA-MB-231 and MDA-MB-468 cells by Sulforhodamine B assay. It exhibited antiproliferative response and induced apoptosis in both the cell types. Cell cycle analysis revealed that it increases the sub G0/G1 phase cell population. Modulation of mitochondrial membrane potential, activation of caspase 3/7 and a remarkable increase in the expression of cleaved PARP and increased chromatin condensation was observed after CHET treatment in MDA-MB-231 and MDA-MB-468 cells. Additionally, an elevated level of intracellular Ca played an important role in CHET mediated cell death response. Calcium overload in mitochondria led to release of cytochrome c which in turn triggered caspase-3 mediated cell death. Inhibition of calcium signalling using BAPTA-AM reduced apoptosis confirming the involvement of calcium signalling in CHET induced cell death. Chetomin also inhibited PI3K/mTOR cell survival pathway in human TNBC cells. The overall findings suggest that Chetomin inhibited the growth of human TNBC cells by caspase-dependent apoptosis and modulation of PI3K/mTOR signalling and could be used as a novel chemotherapeutic agent for the treatment of human TNBC in future.
[Mh] Termos MeSH primário: Apoptose/efeitos dos fármacos
Cálcio/metabolismo
Dissulfetos/administração & dosagem
Alcaloides de Indol/administração & dosagem
Mitocôndrias/efeitos dos fármacos
Mitocôndrias/patologia
Neoplasias de Mama Triplo Negativas/tratamento farmacológico
Neoplasias de Mama Triplo Negativas/metabolismo
[Mh] Termos MeSH secundário: Antineoplásicos/administração & dosagem
Linhagem Celular Tumoral
Relação Dose-Resposta a Droga
Seres Humanos
Resultado do Tratamento
Neoplasias de Mama Triplo Negativas/patologia
[Pt] Tipo de publicação:JOURNAL ARTICLE; RESEARCH SUPPORT, NON-U.S. GOV'T
[Nm] Nome de substância:
0 (Antineoplastic Agents); 0 (Disulfides); 0 (Indole Alkaloids); 1403-36-7 (chetomin); SY7Q814VUP (Calcium)
[Em] Mês de entrada:1802
[Cu] Atualização por classe:180214
[Lr] Data última revisão:
180214
[Sb] Subgrupo de revista:IM
[Da] Data de entrada para processamento:171207
[St] Status:MEDLINE


  9 / 16628 MEDLINE  
              first record previous record next record last record
seleciona
para imprimir
Fotocópia
Texto completo
[PMID]:29253024
[Au] Autor:Gaciarz A; Ruddock LW
[Ad] Endereço:Faculty of Biochemistry and Molecular Medicine, University of Oulu, Oulu, Finland.
[Ti] Título:Complementarity determining regions and frameworks contribute to the disulfide bond independent folding of intrinsically stable scFv.
[So] Source:PLoS One;12(12):e0189964, 2017.
[Is] ISSN:1932-6203
[Cp] País de publicação:United States
[La] Idioma:eng
[Ab] Resumo:CyDisCo is a system facilitating disulfide bond formation in recombinant proteins in the cytoplasm of Escherichia coli. Previously we screened for soluble expression of single chain antibody fragments (scFv) in the cytoplasm of E. coli in the presence and absence of CyDisCo, with >90% being solubly expressed. Two scFv, those derived from natalizumab and trastuzumab, were solubly produced in high amounts even in the absence of folding catalysts i.e. disulfide bond formation is not critical for their folding. Here we investigate the contribution of the framework and the complementarity determining regions (CDRs) of scFv to the disulfide-independence of folding. We swapped CDRs between four scFv that have different properties, including two scFv that can efficiently fold independently from disulfide bonds and two more disulfide-dependent scFv. To confirm disulfide-independence we generated cysteine to alanine mutants of the disulfide-independent scFv. All of the scFv were tested for soluble expression in the cytoplasm of E. coli in the presence and absence of the oxidative folding catalysts Erv1p and PDI. Eight of the hybrid scFv were solubly produced in the presence of CyDisCo, while seven were solubly produced in the absence of CyDisCo, though the yields were often much lower when CyDisCo was absent. Soluble expression was also observed for scFv natalizumab and trastuzumab containing no cysteines. We compared yields, thermal stability and secondary structure of solubly produced scFv and undertook binding studies by western blotting, dot blotting or surface plasmon resonance of those produced in good yields. Our results indicate that both the CDRs and the framework contribute to the disulfide-dependence of soluble production of scFv, with the CDRs having the largest effect. In addition, there was no correlation between thermal stability and disulfide-dependence of folding and only a weak correlation between the yield of protein and the thermal stability of the protein.
[Mh] Termos MeSH primário: Regiões Determinantes de Complementaridade/química
Dissulfetos/química
Proteínas Recombinantes/química
Anticorpos de Cadeia Única/química
[Mh] Termos MeSH secundário: Alanina/genética
Catálise
Escherichia coli/metabolismo
Concentração de Íons de Hidrogênio
Cinética
Mutação
Natalizumab/química
Oxirredução
Oxigênio/química
Dobramento de Proteína
Trastuzumab/química
[Pt] Tipo de publicação:JOURNAL ARTICLE
[Nm] Nome de substância:
0 (Complementarity Determining Regions); 0 (Disulfides); 0 (Natalizumab); 0 (Recombinant Proteins); 0 (Single-Chain Antibodies); OF5P57N2ZX (Alanine); P188ANX8CK (Trastuzumab); S88TT14065 (Oxygen)
[Em] Mês de entrada:1801
[Cu] Atualização por classe:180112
[Lr] Data última revisão:
180112
[Sb] Subgrupo de revista:IM
[Da] Data de entrada para processamento:171219
[St] Status:MEDLINE
[do] DOI:10.1371/journal.pone.0189964


  10 / 16628 MEDLINE  
              first record previous record
seleciona
para imprimir
Fotocópia
[PMID]:27777209
[Au] Autor:Chai JX; Li HH; Wang YY; Chai Q; He WX; Zhou YM; Hu XD; Wang ZH
[Ad] Endereço:1Department of Histology and Embryology, Bengbu Medical College, Bengbu 233030, China; 4Anhui Key Laboratory of Tissue Transplantation, Bengbu 233030, China. E-mail: chaichai123456@163.com.
[Ti] Título:[Effect of diallyl disulfide on learning and memory abilities and hippocampal synapses in mouse models of Alzheimer's disease].
[So] Source:Nan Fang Yi Ke Da Xue Xue Bao;36(10):1417-1422, 2016 Oct 20.
[Is] ISSN:1673-4254
[Cp] País de publicação:China
[La] Idioma:chi
[Ab] Resumo:OBJECTIVE: To explore the effect of diallyl disulfide (DADS) on hippocampal synapses and learning and memory abilities in a mouse model of A1zheimer's disease (AD). METHODS: Mouse models of AD established by agglutinated Aß1-42 injection in the lateral cerebral ventricle were randomized into 4 groups and treated with DADS at the daily doses of 0, 10, 50 and 100 mg/kg by gavage for 30 consecutive days. The learning and memory abilities of the mice were assessed with Morris water maze test; the structures of the dendritic spines and synapses in CA1 region of the hippocampus were observed under transmission electron microscope with silver staining; PSD95 and SYP protein and mRNA expressions in the hippocampus were detected with Western blotting and RT-PCR. RESULTS: Compared with the AD model mice, the mice treated with 50 and 100 mg/kg DADS showed enhanced learning and memory abilities in Morris water maze test. The dendritic spines and synapses in CA1 region of the hippocampus increased obviously and hippocampal expressions of PSD95 and SYP were enhanced in mice treated with 50 and 100 mg/kg DADS. CONCLUSION: DADS at the daily doses of 50 and 100 mg/kg can improve the learning and memory abilities and increase the number of dendritic spines and synapses in the hippocampus in mouse models of AD.
[Mh] Termos MeSH primário: Compostos Alílicos/farmacologia
Doença de Alzheimer/tratamento farmacológico
Dissulfetos/farmacologia
Aprendizagem
Memória
[Mh] Termos MeSH secundário: Animais
Região CA1 Hipocampal/efeitos dos fármacos
Modelos Animais de Doenças
Masculino
Camundongos
Sinapses/efeitos dos fármacos
[Pt] Tipo de publicação:JOURNAL ARTICLE
[Nm] Nome de substância:
0 (Allyl Compounds); 0 (Disulfides); 5HI47O6OA7 (diallyl disulfide)
[Em] Mês de entrada:1711
[Cu] Atualização por classe:171128
[Lr] Data última revisão:
171128
[Sb] Subgrupo de revista:IM
[Da] Data de entrada para processamento:161026
[St] Status:MEDLINE



página 1 de 1663 ir para página                         
   


Refinar a pesquisa
  Base de dados : MEDLINE Formulário avançado   

    Pesquisar no campo  
1  
2
3
 
           



Search engine: iAH v2.6 powered by WWWISIS

BIREME/OPAS/OMS - Centro Latino-Americano e do Caribe de Informação em Ciências da Saúde