Your browser doesn't support javascript.
loading
Mostrar: 20 | 50 | 100
Resultados 1 - 20 de 122
Filtrar
Mais filtros

Base de dados
País/Região como assunto
Tipo de documento
Intervalo de ano de publicação
1.
Nucleic Acids Res ; 51(12): 6495-6506, 2023 07 07.
Artigo em Inglês | MEDLINE | ID: mdl-36919612

RESUMO

5-methylcytosine (mC) and its TET-oxidized derivatives exist in CpG dyads of mammalian DNA and regulate cell fate, but how their individual combinations in the two strands of a CpG act as distinct regulatory signals is poorly understood. Readers that selectively recognize such novel 'CpG duplex marks' could be versatile tools for studying their biological functions, but their design represents an unprecedented selectivity challenge. By mutational studies, NMR relaxation, and MD simulations, we here show that the selectivity of the first designer reader for an oxidized CpG duplex mark hinges on precisely tempered conformational plasticity of the scaffold adopted during directed evolution. Our observations reveal the critical aspect of defined motional features in this novel reader for affinity and specificity in the DNA/protein interaction, providing unexpected prospects for further design progress in this novel area of DNA recognition.


Assuntos
5-Metilcitosina , DNA , Epigênese Genética , Animais , Ilhas de CpG/genética , DNA/química , Metilação de DNA , Epigenômica , Mamíferos/metabolismo , Conformação Molecular
2.
Chembiochem ; 25(11): e202400057, 2024 Jun 03.
Artigo em Inglês | MEDLINE | ID: mdl-38390661

RESUMO

Halophilic organisms have adapted to multi-molar salt concentrations, their cytoplasmic proteins functioning despite stronger attraction between hydrophobic groups. These proteins, of interest in biotechnology because of decreasing fresh-water resources, have excess acidic amino acids. It has been suggested that conformational fluctuations - critical for protein function - decrease in the presence of a stronger hydrophobic effect, and that an acidic proteome would counteract this decrease. However, our understanding of the salt- and acidic amino acid dependency of enzymatic activity is limited. Here, using solution NMR relaxation and molecular dynamics simulations for in total 14 proteins, we show that salt concentration has a limited and moreover non-monotonic impact on protein dynamics. The results speak against the conformational-fluctuations model, instead indicating that maintaining protein dynamics to ensure protein function is not an evolutionary driving force behind the acidic proteome of halophilic proteins.


Assuntos
Simulação de Dinâmica Molecular , Soluções , Eletrólitos/química , Proteínas/química , Proteínas/metabolismo , Interações Hidrofóbicas e Hidrofílicas , Conformação Proteica , Ressonância Magnética Nuclear Biomolecular
3.
Chem Biodivers ; 21(8): e202400642, 2024 Aug.
Artigo em Inglês | MEDLINE | ID: mdl-38822644

RESUMO

New 2-(4-benzothiazol-2-yl-phenoxy)-1-(3,5-diphenyl-4,5-dihydro-pyrazol-1-yl)-ethanones (9a-o) have been designed and synthesized. All the synthesized compounds were characterized by thin layer chromatography and spectral analysis. The antiepileptic potential of the synthesized compounds has been tested by following standard animal screening models, including maximal electroshock (MES) and subcutaneous pentylenetetrazole (scPTZ) models. The neurotoxic and antidepression effects of the synthesized compounds were checked by utilizing rotarod apparatus, and motor impairment test (by actophotometer) respectively. The study concluded that compounds 9c, 9d, 9f, 9i, 9n, and 9o possessed good antiepileptic potential compared to standard drugs like carbamazepine and phenytoin. The results of the rotarod performance test also established them without any neurotoxicity. The motor impairment test revealed that the synthesized compounds are also good antidepressants. In-silico studies have been performed for calculation of pharmacophore pattern, prediction of pharmacokinetic properties which determine the eligibility of synthesized compounds as orally administered molecules and interactions with the target proteins. The result of in-silico studies reinforced results obtained by in vivo study of the synthesized compounds and their possible mechanism of antiepileptic action i. e. via inhibiting voltage-gated sodium channels (VGSCs) and gamma-aminobutyric acid-A receptor.


Assuntos
Anticonvulsivantes , Benzotiazóis , Pirazóis , Anticonvulsivantes/química , Anticonvulsivantes/síntese química , Anticonvulsivantes/farmacologia , Animais , Benzotiazóis/química , Benzotiazóis/antagonistas & inibidores , Benzotiazóis/farmacologia , Benzotiazóis/síntese química , Camundongos , Pirazóis/química , Pirazóis/farmacologia , Pirazóis/síntese química , Pentilenotetrazol , Eletrochoque , Relação Estrutura-Atividade , Convulsões/tratamento farmacológico , Convulsões/induzido quimicamente , Masculino , Estrutura Molecular , Simulação de Acoplamento Molecular , Modelos Animais de Doenças
4.
J Asian Nat Prod Res ; 26(6): 663-680, 2024 Jun.
Artigo em Inglês | MEDLINE | ID: mdl-38373215

RESUMO

Kinase is an enzyme that helps in the phosphorylation of the targeted molecules and can affect their ability to react with other molecules. So, kinase influences metabolic reactions like cell signaling, secretory processes, transport of molecules, etc. The increased activity of certain kinases may cause various types of cancer, i.e. leukemia, glioblastoma, and neuroblastomas. So, the growth of particular cancer cells can be prevented by the inhibition of the kinase responsible for those cancers. Natural products are the key resources for the development of new drugs where approximately 60% of anti-tumor drugs are being developed with the same including specific kinase dwellers. This study comprised molecular interactions of various molecules (obtained from natural sources) as kinase inhibitors for the treatment of cancer. It is expected that by analyzing the skeleton behavior, the process of action, and the body-related activity of these organic products, new cancer-avoiding molecules can be developed.


Assuntos
Antineoplásicos , Produtos Biológicos , Neoplasias , Inibidores de Proteínas Quinases , Humanos , Produtos Biológicos/farmacologia , Produtos Biológicos/química , Inibidores de Proteínas Quinases/farmacologia , Inibidores de Proteínas Quinases/química , Estrutura Molecular , Neoplasias/tratamento farmacológico , Antineoplásicos/farmacologia , Antineoplásicos/química
5.
Angew Chem Int Ed Engl ; 63(5): e202313947, 2024 Jan 25.
Artigo em Inglês | MEDLINE | ID: mdl-37974542

RESUMO

The possible internal dynamics of non-isotope-labeled small-molecule ligands inside a target protein is inherently difficult to capture. Whereas high crystallographic temperature factors can denote either static disorder or motion, even moieties with very low B-factors can be subject to vivid motion between symmetry-related sites. Here we report the experimental identification of internal µs timescale dynamics of a high-affinity, natural-abundance ligand tightly bound to the enzyme human carbonic anhydrase II (hCAII) even within a crystalline lattice. The rotamer jumps of the ligand's benzene group manifest themselves both, in solution and fast magic-angle spinning solid-state NMR 1 H R1ρ relaxation dispersion, for which we obtain further mechanistic insights from molecular-dynamics (MD) simulations. The experimental confirmation of rotameric jumps in bound ligands within proteins in solution or the crystalline state may improve understanding of host-guest interactions in biology and supra-molecular chemistry and may facilitate medicinal chemistry for future drug campaigns.


Assuntos
Proteínas , Humanos , Ressonância Magnética Nuclear Biomolecular , Domínio Catalítico , Ligantes , Proteínas/química , Espectroscopia de Ressonância Magnética
6.
Hum Mol Genet ; 30(18): 1734-1749, 2021 08 28.
Artigo em Inglês | MEDLINE | ID: mdl-34007987

RESUMO

High-altitude (HA, >2500 m) hypoxic exposure evokes several physiological processes that may be abetted by differential genetic distribution in sojourners, who are susceptible to various HA disorders, such as high-altitude pulmonary edema (HAPE). The genetic variants in hypoxia-sensing genes influence the transcriptional output; however the functional role has not been investigated in HAPE. This study explored the two hypoxia-sensing genes, prolyl hydroxylase domain protein 2 (EGLN1) and factor inhibiting HIF-1α (HIF1AN) in HA adaptation and maladaptation in three well-characterized groups: highland natives, HAPE-free controls and HAPE-patients. The two genes were sequenced and subsequently validated through genotyping of significant single nucleotide polymorphisms (SNPs), haplotyping and multifactor dimensionality reduction. Three EGLN1 SNPs rs1538664, rs479200 and rs480902 and their haplotypes emerged significant in HAPE. Blood gene expression and protein levels also differed significantly (P < 0.05) and correlated with clinical parameters and respective alleles. The RegulomeDB annotation exercises of the loci corroborated regulatory role. Allele-specific differential expression was evidenced by luciferase assay followed by electrophoretic mobility shift assay, liquid chromatography with tandem mass spectrometry and supershift assays, which confirmed allele-specific transcription factor (TF) binding of FUS RNA-binding protein (FUS) with rs1538664A, Rho GDP dissociation inhibitor 1 (ARHDGIA) with rs479200T and hypoxia upregulated protein 1 (HYOU1) with rs480902C. Docking simulation studies were in sync for the DNA-TF structural variations. There was strong networking among the TFs that revealed physiological consequences through relevant pathways. The two hydroxylases appear crucial in the regulation of hypoxia-inducible responses.


Assuntos
Doença da Altitude , Loci Gênicos , Hipertensão Pulmonar , Prolina Dioxigenases do Fator Induzível por Hipóxia , Oxigenases de Função Mista , Polimorfismo de Nucleotídeo Único , Edema Pulmonar , Proteínas Repressoras , Células A549 , Altitude , Doença da Altitude/enzimologia , Doença da Altitude/genética , Feminino , Regulação Enzimológica da Expressão Gênica , Humanos , Hipertensão Pulmonar/enzimologia , Hipertensão Pulmonar/genética , Prolina Dioxigenases do Fator Induzível por Hipóxia/biossíntese , Prolina Dioxigenases do Fator Induzível por Hipóxia/genética , Masculino , Oxigenases de Função Mista/biossíntese , Oxigenases de Função Mista/genética , Edema Pulmonar/enzimologia , Edema Pulmonar/genética , Proteínas Repressoras/biossíntese , Proteínas Repressoras/genética , Fatores de Risco
7.
Org Biomol Chem ; 21(23): 4854-4862, 2023 Jun 14.
Artigo em Inglês | MEDLINE | ID: mdl-37249156

RESUMO

A cost-effective and competent approach has been established for the synthesis of pyrimido[4,5-b]quinolines and pyrido[2,3-d]pyrimidines via a multicomponent reaction of 1,3 diketones (dimedone, barbituric acid, and Meldrum's acid), 6-aminouracil and aromatic aldehyde, through mechanochemical synthesis using a ball-mill. This transformation involves a one pot, catalyst-free, and solvent-free pathway to develop the desired products under mild reaction conditions.

8.
J Am Chem Soc ; 144(7): 2987-2993, 2022 02 23.
Artigo em Inglês | MEDLINE | ID: mdl-35157801

RESUMO

5-Methylcytosine (mC) and 5-hydroxymethylcytosine (hmC), the two main epigenetic modifications of mammalian DNA, exist in symmetric and asymmetric combinations in the two strands of CpG dyads. However, revealing such combinations in single DNA duplexes is a significant challenge. Here, we evolve methyl-CpG-binding domains (MBDs) derived from MeCP2 by bacterial cell surface display, resulting in the first affinity probes for hmC/mC CpGs. One mutant has low nanomolar affinity for a single hmC/mC CpG, discriminates against all 14 other modified CpG dyads, and rivals the selectivity of wild-type MeCP2. Structural studies indicate that this protein has a conserved scaffold and recognizes hmC and mC with two dedicated sets of residues. The mutant allows us to selectively address and enrich hmC/mC-containing DNA fragments from genomic DNA backgrounds. We anticipate that this novel probe will be a versatile tool to unravel the function of hmC/mC marks in diverse aspects of chromatin biology.


Assuntos
5-Metilcitosina/análogos & derivados , 5-Metilcitosina/química , DNA/isolamento & purificação , Proteína 2 de Ligação a Metil-CpG/química , Fragmentos de Peptídeos/química , DNA/química , Metilação de DNA , Evolução Molecular Direcionada , Células HEK293 , Humanos , Proteína 2 de Ligação a Metil-CpG/genética , Fragmentos de Peptídeos/genética , Domínios Proteicos
9.
J Med Virol ; 94(4): 1300-1314, 2022 Apr.
Artigo em Inglês | MEDLINE | ID: mdl-34811761

RESUMO

Young age, female sex, absence of comorbidities, and prior infection or vaccination are known epidemiological barriers for contracting the new infection and/or increased disease severity. Demographic trends from the recent coronavirus disease 2019 waves, which are believed to be driven by newer severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) variants, indicate that the aforementioned epidemiological barriers are being breached and a larger number of younger and healthy individuals are developing severe disease. The new SARS-CoV-2 variants have key mutations that can induce significant changes in the virus-host interactions. Recent studies report that, some of these mutations, singly or in a group, enhance key mechanisms, such as binding of the receptor-binding domain (RBD) of the viral spike protein with the angiotensin-converting enzyme 2 (ACE2) receptor in the host-cells, increase the glycosylation of spike protein at the antigenic sites, and enhance the proteolytic cleavage of the spike protein, thus leading to improved host-cell entry and the replication of the virus. The putative changes in the virus-host interactions imparted by the mutations in the RBD sequence can potentially be the reason behind the breach of the observed epidemiological barriers. Susceptibility for contracting SARS-CoV-2 infection and the disease outcomes are known to be influenced by host-cell expressions of ACE2 and other proteases. The new variants can act more efficiently, and even with the lesser availability of the viral entry-receptor and the associated proteases, can have more efficient host-cell entry and greater replication resulting in high viral loads and prolonged viral shedding, widespread tissue-injury, and severe inflammation leading to increased transmissibility and lethality. Furthermore, the accumulating evidence shows that multiple new variants have reduced neutralization by both, natural and vaccine-acquired antibodies, indicating that repeated and vaccine breakthrough infections may arise as serious health concerns in the ongoing pandemic.


Assuntos
COVID-19/epidemiologia , COVID-19/virologia , SARS-CoV-2/fisiologia , SARS-CoV-2/patogenicidade , COVID-19/transmissão , Suscetibilidade a Doenças , Interações Hospedeiro-Patógeno , Humanos , Evasão da Resposta Imune , Mutação , SARS-CoV-2/genética , SARS-CoV-2/imunologia , Glicoproteína da Espícula de Coronavírus/genética , Glicoproteína da Espícula de Coronavírus/imunologia , Internalização do Vírus , Replicação Viral
10.
Cancer Cell Int ; 22(1): 234, 2022 Jul 25.
Artigo em Inglês | MEDLINE | ID: mdl-35879772

RESUMO

Cancer is one of the major causes of mortality worldwide, therefore it is considered a major health concern. Breast cancer is the most frequent type of cancer which affects women on a global scale. Various current treatment strategies have been implicated for breast cancer therapy that includes surgical removal, radiation therapy, hormonal therapy, chemotherapy, and targeted biological therapy. However, constant effort is being made to introduce novel therapies with minimal toxicity. Gene therapy is one of the promising tools, to rectify defective genes and cure various cancers. In recent years, a novel genome engineering technology, namely the clustered regularly interspaced short palindromic repeat (CRISPR)-associated protein-9 (Cas9) has emerged as a gene-editing tool and transformed genome-editing techniques in a wide range of biological domains including human cancer research and gene therapy. This could be attributed to its versatile characteristics such as high specificity, precision, time-saving and cost-effective methodologies with minimal risk. In the present review, we highlight the role of CRISPR/Cas9 as a targeted therapy to tackle drug resistance, improve immunotherapy for breast cancer.

11.
Am J Respir Cell Mol Biol ; 65(5): 521-531, 2021 11.
Artigo em Inglês | MEDLINE | ID: mdl-34126864

RESUMO

Stem cells, including the resident lung mesenchymal stem cells (LMSCs), are critically important for injury repair. Compelling evidence links perinatal vitamin D (VD) deficiency to reactive airway disease; however, the effects of perinatal VD deficiency on LMSC function is unknown. We tested the hypothesis that perinatal VD deficiency alters LMSC proliferation, differentiation, and function, leading to an enhanced myogenic phenotype. We also determined whether LMSCs' effects on alveolar type II (ATII) cell function are paracrine. Using an established rat model of perinatal VD deficiency, we studied the effects of four dietary regimens (0, 250, 500, or 1,000 IU/kg cholecalciferol-supplemented groups). At Postnatal Day 21, LMSCs were isolated, and cell proliferation and differentiation (under basal and adipogenic induction conditions) were determined. LMSC paracrine effects on ATII cell proliferation and differentiation were determined by culturing ATII cells in LMSC-conditioned media from different experimental groups. Using flow cytometry, >95% of cells were CD45-ve, >90% were CD90 + ve, >58% were CD105 + ve, and >64% were Stro-1 + ve, indicating their stem cell phenotype. Compared with the VD-supplemented groups, LMSCs from the VD-deficient group demonstrated suppressed PPARγ, but enhanced Wnt signaling, under basal and adipogenic induction conditions. LMSCs from 250 VD- and 500 VD-supplemented groups effectively blocked the effects of perinatal VD deficiency. LMSC-conditioned media from the VD-deficient group inhibited ATII cell proliferation and differentiation compared with those from the 250 VD- and 500 VD-supplemented groups. These data support the concept that perinatal VD deficiency alters LMSC proliferation and differentiation, potentially contributing to increased respiratory morbidity seen in children born to mothers with VD deficiency.


Assuntos
Pulmão/citologia , Células-Tronco Mesenquimais/citologia , Deficiência de Vitamina D/complicações , Adipócitos/citologia , Adipócitos/efeitos dos fármacos , Animais , Diferenciação Celular/efeitos dos fármacos , Proliferação de Células , Células Cultivadas , Relação Dose-Resposta a Droga , Feminino , Pulmão/fisiologia , Pulmão/fisiopatologia , Células-Tronco Mesenquimais/efeitos dos fármacos , Células-Tronco Mesenquimais/fisiologia , Desenvolvimento Muscular/efeitos dos fármacos , Desenvolvimento Muscular/fisiologia , Gravidez , Ratos , Vitamina D/administração & dosagem , Vitamina D/farmacologia , Via de Sinalização Wnt
12.
Mol Divers ; 25(1): 131-142, 2021 Feb.
Artigo em Inglês | MEDLINE | ID: mdl-31919739

RESUMO

A facile and efficient multicomponent synthesis of benzodiazepine ring in water under ultrasound irradiation is reported first time. The current procedure escapes traditional chromatography and purification process and provided the product in excellent yields of 95% as compared to conventional methods. The approach was also validated on gram-scale synthesis.


Assuntos
Benzodiazepinas/química , Benzodiazepinas/síntese química , Água/química , Ondas Ultrassônicas
13.
Org Biomol Chem ; 18(13): 2492-2500, 2020 04 01.
Artigo em Inglês | MEDLINE | ID: mdl-32186577

RESUMO

1,3-Bis(1-alkyl-1H-indol-3-yl)benzene derivatives have been synthesized through a Zn(OTf)2 catalyzed reaction between cyclohexanones and indoles. Cyclohexanones act as an aryl source via an alkylation-dehydrogenation-aromatization sequence, and undergo regioselective C-C bond formation with indoles at the 1,3-position. A useful library of these derivatives obtained in moderate to high yields has been prepared and a tentative mechanism has been proposed based upon GCMS analysis and time dependent NMR studies.

14.
Angew Chem Int Ed Engl ; 59(51): 22916-22921, 2020 12 14.
Artigo em Inglês | MEDLINE | ID: mdl-32965765

RESUMO

Drug discovery, in particular optimization of candidates using medicinal chemistry, is generally guided by structural biology. However, for optimizing binding kinetics, relevant for efficacy and off-target effects, information on protein motion is important. Herein, we demonstrate for the prototypical textbook example of an allegedly "rigid protein" that substantial active-site dynamics have generally remained unrecognized, despite thousands of medicinal-chemistry studies on this model over decades. Comparing cryogenic X-ray structures, solid-state NMR on micro-crystalline protein at room temperature, and solution NMR structure and dynamics, supported by MD simulations, we show that under physiologically relevant conditions the pocket is in fact shaped by pronounced open/close conformational-exchange dynamics. The study, which is of general significance for pharmacological research, evinces a generic pitfall in drug discovery routines.


Assuntos
Simulação de Dinâmica Molecular , Preparações Farmacêuticas/química , Proteínas/química , Sítios de Ligação , Ressonância Magnética Nuclear Biomolecular , Conformação Proteica
15.
Malays J Med Sci ; 27(6): 53-67, 2020 Dec.
Artigo em Inglês | MEDLINE | ID: mdl-33447134

RESUMO

BACKGROUND: Ischaemic stroke (IS), a multifactorial neurological disorder, is mediated by interplay between genes and the environment and, thus, blood-based IS biomarkers are of significant clinical value. Therefore, this study aimed to find global differentially expressed genes (DEGs) in-silico, to identify key enriched genes via gene set enrichment analysis (GSEA) and to determine the clinical significance of these genes in IS. METHODS: Microarray expression dataset GSE22255 was retrieved from the Gene Expression Omnibus (GEO) database. It includes messenger ribonucleic acid (mRNA) expression data for the peripheral blood mononuclear cells of 20 controls and 20 IS patients. The bioconductor-package 'affy' was used to calculate expression and a pairwise t-test was applied to screen DEGs (P < 0.01). Further, GSEA was used to determine the enrichment of DEGs specific to gene ontology (GO) annotations. RESULTS: GSEA analysis revealed 21 genes to be significantly plausible gene markers, enriched in multiple pathways among all the DEGs (n = 881). Ten gene sets were found to be core enriched in specific GO annotations. JunD, NCX3 and fibroblast growth factor receptor 4 (FGFR4) were under-represented and glycoprotein M6-B (GPM6B) was persistently over-represented. CONCLUSION: The identified genes are either associated with the pathophysiology of IS or they affect post-IS neuronal regeneration, thereby influencing clinical outcome. These genes should, therefore, be evaluated for their utility as suitable markers for predicting IS in clinical scenarios.

16.
J Am Chem Soc ; 141(49): 19276-19288, 2019 12 11.
Artigo em Inglês | MEDLINE | ID: mdl-31647225

RESUMO

Protein-water interactions have widespread effects on protein structure and dynamics. As such, the function of many biomacromolecules can be directly related to the presence and exchange of water molecules. While the presence of structural water sites can be easily detected by X-ray crystallography, the dynamics within functional water-protein network architectures is largely elusive. Here we use solid-state NMR relaxation dispersion measurements with a focus on those active-site residues in the enzyme human carbonic anhydrase II (hCAII) that constitute the evolutionarily conserved water pocket, key for CAs' enzymatic catalysis. Together with chemical shifts, peak broadening, and results of molecular dynamics (MD) and DFT shift calculations, the relaxation dispersion data suggest the presence of a widespread fast µs-time-scale dynamics in the pocket throughout the protein-water network. This process is abrogated in the presence of an inhibitor which partially disrupts the network. The time scale of the protein-water pocket motion coincides both with the estimated residence time of Zn-bound water/OH- in the pocket showing the longest lifetimes in earlier magnetic relaxation dispersion experiments as well as with the rate-limiting step of catalytic turnover. As such, the reorganization of the water pocket:enzyme architecture might constitute an element of importance for enzymatic activity of this and possibly other proteins.


Assuntos
Anidrase Carbônica II/química , Simulação de Dinâmica Molecular , Ressonância Magnética Nuclear Biomolecular , Água/química , Anidrase Carbônica II/genética , Domínio Catalítico , Escherichia coli/genética , Humanos , Ligação de Hidrogênio , Ligação Proteica , Conformação Proteica
17.
Anal Biochem ; 570: 62-64, 2019 04 01.
Artigo em Inglês | MEDLINE | ID: mdl-30735667

RESUMO

We report the observation of single-site phosphorylation in a His-tag sequence N-terminally attached to a recombinant protein (UVI31+) in vitro. This modification was detected at position 23 at a serine residue of the His-tag sequence encoded by the vector pET28a. Furthermore, the phosphorylated tag sequence was found to be dephosphorylated by the action of alkaline phosphatases. The functional activity and dynamics of the protein carrying the His-tag sequence were unchanged after phosphorylation. The possibility of phosphorylation within the N-terminal His-tag demonstrates that care has to be taken upon analysis of post-translational modifications via mass spectrometry for recombinant protein expression strategies.


Assuntos
Proteínas Recombinantes de Fusão/análise , Espectrometria de Massas por Ionização e Dessorção a Laser Assistida por Matriz , Fosfatase Alcalina/metabolismo , Histidina/genética , Histidina/metabolismo , Isótopos de Nitrogênio/química , Ressonância Magnética Nuclear Biomolecular , Oligopeptídeos/genética , Oligopeptídeos/metabolismo , Fosfopeptídeos/análise , Fosforilação , Proteínas Recombinantes de Fusão/metabolismo
18.
Angew Chem Int Ed Engl ; 58(17): 5758-5762, 2019 04 16.
Artigo em Inglês | MEDLINE | ID: mdl-30688395

RESUMO

Solid-state NMR spectroscopy has recently enabled structural biology with small amounts of non-deuterated proteins, largely alleviating the classical sample production demands. Still, despite the benefits for sample preparation, successful and comprehensive characterization of complex spin systems in the few cases of higher-molecular-weight proteins has thus far relied on traditional 13 C-detected methodology or sample deuteration. Herein we show for a 29 kDa carbonic anhydrase:acetazolamide complex that different aspects of solid-state NMR assessment of a complex spin system can be successfully accessed using a non-deuterated, 500 µg sample in combination with adequate spectroscopic tools. The shown access to protein structure, protein dynamics, as well as biochemical parameters in amino acid sidechains, such as histidine protonation states, will be transferable to proteins that are not expressible in E. coli.


Assuntos
Deutério/química , Espectroscopia de Ressonância Magnética/métodos , Humanos , Prótons
19.
Cell Microbiol ; 19(6)2017 06.
Artigo em Inglês | MEDLINE | ID: mdl-28030753

RESUMO

The extensive modification of Plasmodium falciparum-infected erythrocytes by variant surface antigens plays a major role in immune evasion and malaria-induced pathology. Here, using high-resolution microscopy, we visualize the spatio-temporal expression dynamics of STEVOR, an important variant surface antigens family, in a stage-dependent manner. We demonstrate that it is exported to the cell surface where protein molecules cluster and preferentially localize in proximity to knobs. Quantitative evidence from our force measurements and microfluidic assays reveal that STEVOR can effectively mediate the formation of stable, robust rosettes under static and physiologically relevant flow conditions. Our results extend previously published studies in P. falciparum and emphasize the role of STEVOR in rosetting, an important contributor to disease pathology.


Assuntos
Antígenos de Protozoários/genética , Antígenos de Superfície/genética , Adesão Celular/genética , Plasmodium falciparum/patogenicidade , Proteínas de Protozoários/genética , Antígenos de Protozoários/biossíntese , Antígenos de Superfície/biossíntese , Adesão Celular/fisiologia , Linhagem Celular , Eritrócitos/parasitologia , Humanos , Malária Falciparum/parasitologia , Plasmodium falciparum/genética , Plasmodium falciparum/metabolismo , Proteínas de Protozoários/biossíntese , Formação de Roseta
20.
J Minim Access Surg ; 14(3): 192-196, 2018.
Artigo em Inglês | MEDLINE | ID: mdl-29067946

RESUMO

PREMISE AND OBJECTIVE:: Elective laparoscopic cholecystectomy (LC) has low risk for post-operative infectious complications; still most clinicians use persistent post-operative prophylactic antibiotics out of habit, tradition, or simply as defensive practice due to evolving medicolegal implications of a large number of surgeries being showcased as daycare or next day discharge procedures. This randomised prospective trial was done to test the need for such prophylaxis in cases of elective LC in a rural/semi-urban setting. MATERIALS AND METHODS:: Two hundred and ten successive patients undergoing elective LC were randomised into groups receiving single dose of injection ceftriaxone at the time of induction of anaesthesia, (Group A = 112 cases) and those who in addition to above received injection ceftriaxone twice daily for 2 days postoperatively (Group B = 98 cases). Post-operative infectious complications between two groups were compared for variables such as age, sex, body mass index and bile/stone spillage. RESULTS:: There was no significant difference in surgical site infection rates between the groups for variables such as age, sex, body mass index, duration of symptoms, American Society of Anesthesiologists grade, duration of surgery and hospital stay. Intraoperative spillage of stones (9.8% [A]: 5.1% [B]) did not increase infectious complications even in the presence of positive bile culture (Group A, N = 7 vs. Group B, N = 3). An operative time of greater than 60 min was found to be associated with increased surgical site infection (P = 0. 0006). CONCLUSION:: Single dose of ceftriaxone at the time of induction is adequate prophylaxis following elective LC even in the rural/semi-urban Indian setting and routine continued administration of antibiotic should be abandoned as it contributes to adverse reactions, drug resistance and unnecessary financial burden.

SELEÇÃO DE REFERÊNCIAS
DETALHE DA PESQUISA