共查询到20条相似文献,搜索用时 15 毫秒
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Monitoring the Glutathione Redox Reaction in Living Human Cells by Combining Metabolic Labeling with Heteronuclear NMR 下载免费PDF全文
Xing Jin Soeun Kang Prof. Shinya Tanaka Prof. Sunghyouk Park 《Angewandte Chemie (International ed. in English)》2016,55(28):7939-7942
The glutathione (GSH) redox reaction is critical for defense against cellular reactive oxygen species (ROS). However, direct and real‐time monitoring of this reaction in living mammalian cells has been hindered by the lack of a facile method. Herein, we describe a new approach that exploits the GSH biosynthetic pathway and heteronuclear NMR. [U‐13C]‐labeled cysteine was incorporated into GSH in U87 glioblastoma cells, and the oxidation of GSH to GSSG by a ROS‐producing agent could be monitored in living cells. Further application of the approach to cells resistant to temozolomide (TMZ), an anti‐glioblastoma drug, suggested a possible new resistance mechanism involving neutralization of ROS. This result was corroborated by the observation of up‐regulation of glutathione peroxidase 3 (GPx3). This new approach could be easily applied to redox‐dependent signaling pathways and drug resistance involving ROS. 相似文献
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Robert Hänsel Laura M. Luh Ivan Corbeski Lukáš Trantirek Prof. Volker Dötsch 《Angewandte Chemie (International ed. in English)》2014,53(39):10300-10314
The dream of cell biologists is to be able to watch biological macromolecules perform their duties in the intracellular environment of live cells. Ideally, the observation of both the location and the conformation of these macromolecules with biophysical techniques is desired. The development of many fluorescence techniques, including superresolution fluorescence microscopy, has significantly enhanced our ability to spot proteins and other molecules in the crowded cellular environment. However, the observation of their structure and conformational changes while they attend their business is still very challenging. In principle, NMR and EPR spectroscopy can be used to investigate the conformation and dynamics of biological macromolecules in living cells. The development of in‐cell magnetic resonance techniques has demonstrated the feasibility of this approach. Herein we review the different techniques with a focus on liquid‐state in‐cell NMR spectroscopy, provide an overview of applications, and discuss the challenges that lie ahead. 相似文献
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Real‐Time Monitoring of Cancer Cell Metabolism and Effects of an Anticancer Agent using 2D In‐Cell NMR Spectroscopy 下载免费PDF全文
Dr. He Wen Yong Jin An Wen Jun Xu Prof. Keon Wook Kang Prof. Sunghyouk Park 《Angewandte Chemie (International ed. in English)》2015,54(18):5374-5377
Altered metabolism is a critical part of cancer cell properties, but real‐time monitoring of metabolomic profiles has been hampered by the lack of a facile method. Here, we propose real‐time metabolomic monitoring of live cancer cells using 13C6‐glucose and heteronuclear two‐dimensional (2D) NMR. The method allowed for metabolomic differentiation between cancer and normal cells on the basis of time‐dependent changes in metabolite concentrations. Cancer cells were found to have large in‐ and out‐flux of pyruvate as well as increased net production of alanine and acetate. The method also enabled evaluation of the metabolic effects of galloflavin whose anticancer effects have been attributed to its specific inhibition of lactate dehydrogenase. Our approach revealed previously unknown functional targets of galloflavin, which were further confirmed at the protein levels. Our method is readily applicable to the study of metabolic alterations in other cellular disease model systems. 相似文献
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Liang Qiao Hongyan Bi Jean‐Marc Busnel Dr. Jerome Waser Prof. Pengyuan Yang Prof. Hubert H. Girault Prof. Baohong Liu Prof. 《Chemistry (Weinheim an der Bergstrasse, Germany)》2009,15(27):6711-6717
In‐source oxidation : In‐source photocatalytic redox reactions for inducing peptide fragmentation are achieved on a TiO2‐derived target plate during laser desorption ionization mass spectrometry in the presence of samples and glucose acting as both an electron donor and a hole conductor (see scheme).
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Evidence of CuI/CuII Redox Process by X‐ray Absorption and EPR Spectroscopy: Direct Synthesis of Dihydrofurans from β‐Ketocarbonyl Derivatives and Olefins 下载免费PDF全文
Hong Yi Zhixiong Liao Guanghui Zhang Guoting Zhang Chao Fan Xu Zhang Prof. Dr. Emilio E. Bunel Dr. Chih‐Wen Pao Prof. Dr. Jyh‐Fu Lee Prof. Dr. Aiwen Lei 《Chemistry (Weinheim an der Bergstrasse, Germany)》2015,21(52):18925-18929
The CuI/CuII and CuI/CuIII catalytic cycles have been subject to intense debate in the field of copper‐catalyzed oxidative coupling reactions. A mechanistic study on the CuI/CuII redox process, by X‐ray absorption (XAS) and electron paramagnetic resonance (EPR) spectroscopies, has elucidated the reduction mechanism of CuII to CuI by 1,3‐diketone and detailed investigation revealed that the halide ion is important for the reduction process. The oxidative nature of the thereby‐formed CuI has also been studied by XAS and EPR spectroscopy. This mechanistic information is applicable to the copper‐catalyzed oxidative cyclization of β‐ketocarbonyl derivatives to dihydrofurans. This protocol provides an ideal route to highly substituted dihydrofuran rings from easily available 1,3‐dicarbonyls and olefins. 相似文献
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Jakob T. Nielsen Dr. Morten Bjerring Dr. Martin D. Jeppesen Ronnie O. Pedersen Jan M. Pedersen Dr. Kim L. Hein Thomas Vosegaard Dr. Troels Skrydstrup Prof. Daniel E. Otzen Prof. Niels C. Nielsen Prof. 《Angewandte Chemie (International ed. in English)》2009,48(12):2118-2121
The fibril structure formed by the amyloidogenic fragment SNNFGAILSS of the human islet amyloid polypeptide (hIAPP) is determined with 0.52 Å resolution. Symmetry information contained in the easily obtainable resonance assignments from solid‐state NMR spectra (see picture), along with long‐range constraints, can be applied to uniquely identify the supramolecular organization of fibrils.
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Islam Alshamleh Nina Krause Christian Richter Nina Kurrle Hubert Serve Ulrich L. Günther Harald Schwalbe 《Angewandte Chemie (International ed. in English)》2020,59(6):2304-2308
Current metabolomics approaches utilize cellular metabolite extracts, are destructive, and require high cell numbers. We introduce here an approach that enables the monitoring of cellular metabolism at lower cell numbers by observing the consumption/production of different metabolites over several kinetic data points of up to 48 hours. Our approach does not influence cellular viability, as we optimized the cellular matrix in comparison to other materials used in a variety of in‐cell NMR spectroscopy experiments. We are able to monitor real‐time metabolism of primary patient cells, which are extremely sensitive to external stress. Measurements are set up in an interleaved manner with short acquisition times (approximately 7 minutes per sample), which allows the monitoring of up to 15 patient samples simultaneously. Further, we implemented our approach for performing tracer‐based assays. Our approach will be important not only in the metabolomics fields, but also in individualized diagnostics. 相似文献
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Samuel F. Cousin Dr. Pavel Kadeřávek Baptiste Haddou Dr. Cyril Charlier Thorsten Marquardsen Jean‐Max Tyburn Dr. Pierre‐Alain Bovier Dr. Frank Engelke Dr. Werner Maas Prof. Dr. Geoffrey Bodenhausen Dr. Philippe Pelupessy Prof. Dr. Fabien Ferrage 《Angewandte Chemie (International ed. in English)》2016,55(34):9886-9889
Nuclear magnetic resonance (NMR) studies have benefited tremendously from the steady increase in the strength of magnetic fields. Spectacular improvements in both sensitivity and resolution have enabled the investigation of molecular systems of rising complexity. At very high fields, this progress may be jeopardized by line broadening, which is due to chemical exchange or relaxation by chemical shift anisotropy. In this work, we introduce a two‐field NMR spectrometer designed for both excitation and observation of nuclear spins in two distinct magnetic fields in a single experiment. NMR spectra of several small molecules as well as a protein were obtained, with two dimensions acquired at vastly different magnetic fields. Resonances of exchanging groups that are broadened beyond recognition at high field can be sharpened to narrow peaks in the low‐field dimension. Two‐field NMR spectroscopy enables the measurement of chemical shifts at optimal fields and the study of molecular systems that suffer from internal dynamics, and opens new avenues for NMR spectroscopy at very high magnetic fields. 相似文献
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Insights into the Crystallization and Structural Evolution of Glycine Dihydrate by In Situ Solid‐State NMR Spectroscopy 下载免费PDF全文
Dr. Paolo Cerreia Vioglio Dr. Giulia Mollica Marie Juramy Dr. Colan E. Hughes Dr. P. Andrew Williams Dr. Fabio Ziarelli Prof. Stéphane Viel Dr. Pierre Thureau Prof. Kenneth D. M. Harris 《Angewandte Chemie (International ed. in English)》2018,57(22):6619-6623
In situ solid‐state NMR spectroscopy is exploited to monitor the structural evolution of a glycine/water glass phase formed on flash cooling an aqueous solution of glycine, with a range of modern solid‐state NMR methods applied to elucidate structural properties of the solid phases present. The glycine/water glass is shown to crystallize into an intermediate phase, which then transforms to the β polymorph of glycine. Our in situ NMR results fully corroborate the identity of the intermediate crystalline phase as glycine dihydrate, which was first proposed only very recently. 相似文献
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Understanding J‐Modulation during Spatial Encoding for Sensitivity‐Optimized Ultrafast NMR Spectroscopy 下载免费PDF全文
Boris Gouilleux Laetitia Rouger Benoît Charrier Dr. Ilya Kuprov Prof. Serge Akoka Dr. Jean‐Nicolas Dumez Dr. Patrick Giraudeau 《Chemphyschem》2015,16(14):3093-3100
Ultrafast (UF) NMR spectroscopy is an approach that yields 2D spectra in a single scan. This methodology has become a powerful analytical tool that is used in a large array of applications. However, UF NMR spectroscopy still suffers from an intrinsic low sensitivity, and from the need to compromise between sensitivity, spectral width, and resolution. In particular, the modulation of signal intensities by the spin–spin J‐coupling interaction (J‐modulation) impacts significantly on the intensities of the spectral peaks. This effect can lead to large sensitivity losses and even to missing spectral peaks, depending on the nature of the spin system. Herein, a general simulation package (Spinach) is used to describe J‐modulation effects in UF experiments. The results from simulations match with experimental data and the results of product operator calculations. Several methods are proposed to optimize the sensitivity in UF COSY spectra. The potential and drawbacks of the different strategies are also discussed. These approaches provide a way to adjust the sensitivity of UF experiments for a large range of applications. 相似文献
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Adil Safeer Fleur Kleijburg Dr. Salima Bahri David Beriashvili Dr. Edwin J. A. Veldhuizen Jacq van Neer Dr. Martin Tegelaar Prof. Dr. Hans de Cock Prof. Dr. Han A. B. Wösten Prof. Dr. Marc Baldus 《Chemistry (Weinheim an der Bergstrasse, Germany)》2023,29(1):e202202616
Solid-state NMR (ssNMR) spectroscopy facilitates the non-destructive characterization of structurally heterogeneous biomolecules in their native setting, for example, comprising proteins, lipids and polysaccharides. Here we demonstrate the utility of high and ultra-high field 1H-detected fast MAS ssNMR spectroscopy, which exhibits increased sensitivity and spectral resolution, to further elucidate the atomic-level composition and structural arrangement of the cell wall of Schizophyllum commune, a mushroom-forming fungus from the Basidiomycota phylum. These advancements allowed us to reveal that Cu(II) ions and the antifungal peptide Cathelicidin-2 mainly bind to cell wall proteins at low concentrations while glucans are targeted at high metal ion concentrations. In addition, our data suggest the presence of polysaccharides containing N-acetyl galactosamine (GalNAc) and proteins, including the hydrophobin proteins SC3, shedding more light on the molecular make-up of cells wall as well as the positioning of the polypeptide layer. Obtaining such information may be of critical relevance for future research into fungi in material science and biomedical contexts. 相似文献
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João Medeiros‐Silva Deni Mance Mark Daniëls Shehrazade Jekhmane Dr. Klaartje Houben Prof. Marc Baldus Dr. Markus Weingarth 《Angewandte Chemie (International ed. in English)》2016,55(43):13606-13610
1H detection can significantly improve solid‐state NMR spectral sensitivity and thereby allows studying more complex proteins. However, the common prerequisite for 1H detection is the introduction of exchangeable protons in otherwise deuterated proteins, which has thus far significantly hampered studies of partly water‐inaccessible proteins, such as membrane proteins. Herein, we present an approach that enables high‐resolution 1H‐detected solid‐state NMR (ssNMR) studies of water‐inaccessible proteins, and that even works in highly complex environments such as cellular surfaces. In particular, the method was applied to study the K+ channel KcsA in liposomes and in situ in native bacterial cell membranes. We used our data for a dynamic analysis, and we show that the selectivity filter, which is responsible for ion conduction and highly conserved in K+ channels, undergoes pronounced molecular motion. We expect this approach to open new avenues for biomolecular ssNMR. 相似文献
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Exploring the Molecular Structure of Imidazolium–Silica‐Based Nanoparticle Networks by Combining Solid‐State NMR Spectroscopy and First‐Principles Calculations 下载免费PDF全文
Dr. Marie‐Alexandra Neouze Martin Kronstein Dr. Marco Litschauer Dr. Michael Puchberger Dr. Cristina Coelho Prof. Christian Bonhomme Prof. Christel Gervais Dr. Frederik Tielens 《Chemistry (Weinheim an der Bergstrasse, Germany)》2014,20(46):15188-15196
A DFT‐based molecular model for imidazolium–silica‐based nanoparticle networks (INNs) is presented. The INNs were synthesized and characterized by using small‐angle X‐ray scattering (SAXS), NMR spectroscopy, and theoretical ab initio calculations. 11B and 31P HETCOR CP MAS experiments were recorded. Calculated 19F NMR spectroscopy results, combined with the calculated anion–imidazolium (IM) distances, predicted the IM chain density in the INN, which was also confirmed from thermogravimetric analysis/mass spectrometry results. The presence of water molecules trapped between the nanoparticles is also suggested. First considerations on possible π–π stacking between the IM rings are presented. The predicted electronic properties confirm the photoluminescence emissions in the correct spectral domain. 相似文献
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Dr. Yannick Coppel Dr. Grégory Spataro Dr. Carole Pagès Dr. Bruno Chaudret Dr. André Maisonnat Dr. Myrtil L. Kahn 《Chemistry (Weinheim an der Bergstrasse, Germany)》2012,18(17):5384-5393
Full NMR characterization of ZnO nanoparticles (NPs) stabilized by various amines (hexadecylamine, dodecylamine, and octylamine) in C7D8 demonstrated that the surface of this apparently simple system was very complex. Using different NMR spectroscopic techniques (1H, PGSE‐NMR, diffusion‐filtered 1H NMR, NOESY, ROESY), we observed at least three different modes of interaction of the amines at the surface of the NPs, in thermodynamic equilibrium with the free amines, the relative populations of which varied with their concentration. The first mode corresponded to a strong interaction between a small amount of amine and the ZnO NPs (kdesorp≈13 s?1). The second mode corresponded to a weak interaction between the amines and the surface of the ZnO NPs (koff(2)≈50–60 s?1). The third, and weakest, mode of interaction corresponded to the formation of a second ligand shell by the amine around the NPs that was held together through van der Waals interactions (koff(1)≈25×105 s?1). The second and third modes were in fast exchange on the NMR timescales with the free amines. The strongly interacting amines at the NPs surface (first mode) were in slow exchange with the other modes. A complex hydrogen‐bonding network at the NPs surface was also observed, which did not only involve the coordinated amine but also THF and water molecules that remained from the synthesis. 相似文献