Your browser doesn't support javascript.
loading
Mostrar: 20 | 50 | 100
Resultados 1 - 20 de 63
Filtrar
1.
Proc Natl Acad Sci U S A ; 119(2)2022 01 11.
Artículo en Inglés | MEDLINE | ID: mdl-34996869

RESUMEN

NMR-assisted crystallography-the integrated application of solid-state NMR, X-ray crystallography, and first-principles computational chemistry-holds significant promise for mechanistic enzymology: by providing atomic-resolution characterization of stable intermediates in enzyme active sites, including hydrogen atom locations and tautomeric equilibria, NMR crystallography offers insight into both structure and chemical dynamics. Here, this integrated approach is used to characterize the tryptophan synthase α-aminoacrylate intermediate, a defining species for pyridoxal-5'-phosphate-dependent enzymes that catalyze ß-elimination and replacement reactions. For this intermediate, NMR-assisted crystallography is able to identify the protonation states of the ionizable sites on the cofactor, substrate, and catalytic side chains as well as the location and orientation of crystallographic waters within the active site. Most notable is the water molecule immediately adjacent to the substrate ß-carbon, which serves as a hydrogen bond donor to the ε-amino group of the acid-base catalytic residue ßLys87. From this analysis, a detailed three-dimensional picture of structure and reactivity emerges, highlighting the fate of the L-serine hydroxyl leaving group and the reaction pathway back to the preceding transition state. Reaction of the α-aminoacrylate intermediate with benzimidazole, an isostere of the natural substrate indole, shows benzimidazole bound in the active site and poised for, but unable to initiate, the subsequent bond formation step. When modeled into the benzimidazole position, indole is positioned with C3 in contact with the α-aminoacrylate Cß and aligned for nucleophilic attack. Here, the chemically detailed, three-dimensional structure from NMR-assisted crystallography is key to understanding why benzimidazole does not react, while indole does.


Asunto(s)
Alanina/análogos & derivados , Dominio Catalítico , Cristalografía por Rayos X/métodos , Espectroscopía de Resonancia Magnética/métodos , Triptófano Sintasa/química , Catálisis , Indoles , Imagen por Resonancia Magnética , Resonancia Magnética Nuclear Biomolecular , Fosfato de Piridoxal/metabolismo , Triptófano Sintasa/metabolismo
2.
J Am Chem Soc ; 146(34): 23663-23668, 2024 Aug 28.
Artículo en Inglés | MEDLINE | ID: mdl-38980938

RESUMEN

The interactions between glycosaminoglycans (GAGs) and proteins are essential in numerous biochemical processes that involve ion-pair interactions. However, there is no evidence of direct and specific interactions between GAGs and collagen proteins in native cartilage. The resolution of solid-state NMR (ssNMR) can offer such information but the detection of GAG interactions in cartilage is limited by the sensitivity of the experiments when 13C and 15N isotopes are at natural abundance. In this communication, this limitation is overcome by taking advantage of dynamic nuclear polarization (DNP)-enhanced magic-angle spinning (MAS) experiments to obtain two-dimensional (2D) 15N-13C and 13C-13C correlations on native samples at natural abundance. These experiments unveiled inter-residue correlations in the aliphatic regions of the collagen protein previously unobserved. Additionally, our findings provide direct evidence of charge-pair salt-bridge interactions between negatively charged GAGs and positively charged arginine (Arg) residues of collagen protein. We also identified potential hydrogen bonding interactions between hydroxyproline (Hyp) and GAGs, offering atomic insights into the biochemical interactions within the extracellular matrix of native cartilage. Our approach may provide a new avenue for the structural characterization of other native systems.


Asunto(s)
Cartílago , Colágeno , Glicosaminoglicanos , Resonancia Magnética Nuclear Biomolecular , Colágeno/química , Colágeno/metabolismo , Glicosaminoglicanos/química , Glicosaminoglicanos/metabolismo , Cartílago/metabolismo , Cartílago/química , Animales , Hidroxiprolina/química , Enlace de Hidrógeno , Sales (Química)/química
3.
J Am Chem Soc ; 146(6): 4187-4211, 2024 Feb 14.
Artículo en Inglés | MEDLINE | ID: mdl-38316011

RESUMEN

Although Bu3Sn-mediated radical alkyne peri-annulations allow access to phenalenyl ring systems, the oxidative termination of these cascades provides only a limited selection of the possible isomeric phenalenone products with product selectivity controlled by the intrinsic properties of the new cyclic systems. In this work, we report an oxidant-free termination strategy that can overcome this limitation and enable selective access to the full set of isomerically functionalized phenalenones. The key to preferential termination is the preinstallation of a "weak link" that undergoes C-O fragmentation in the final cascade step. Breaking a C-O bond is assisted by entropy, gain of conjugation in the product, and release of stabilized radical fragments. This strategy is expanded to radical exo-dig cyclization cascades of oligoalkynes, which provide access to isomeric π-extended phenalenones. Conveniently, these cascades introduce functionalities (i.e., Bu3Sn and iodide moieties) amenable to further cross-coupling reactions. Consequently, a variety of polyaromatic diones, which could serve as phenalenyl-based open-shell precursors, can be synthesized.

4.
J Biomol NMR ; 78(2): 95-108, 2024 Jun.
Artículo en Inglés | MEDLINE | ID: mdl-38520488

RESUMEN

With the sensitivity enhancements conferred by dynamic nuclear polarization (DNP), magic angle spinning (MAS) solid state NMR spectroscopy experiments can attain the necessary sensitivity to detect very low concentrations of proteins. This potentially enables structural investigations of proteins at their endogenous levels in their biological contexts where their native stoichiometries with potential interactors is maintained. Yet, even with DNP, experiments are still sensitivity limited. Moreover, when an isotopically-enriched target protein is present at physiological levels, which typically range from low micromolar to nanomolar concentrations, the isotope content from the natural abundance isotopes in the cellular milieu can outnumber the isotope content of the target protein. Using isotopically enriched yeast prion protein, Sup35NM, diluted into natural abundance yeast lysates, we optimized sample composition. We found that modest cryoprotectant concentrations and fully protonated environments support efficient DNP. We experimentally validated theoretical calculations of the limit of specificity for an isotopically enriched protein in natural abundance cellular milieu. We establish that, using pulse sequences that are selective for adjacent NMR-active nuclei, proteins can be specifically detected in cellular milieu at concentrations in the hundreds of nanomolar. Finally, we find that maintaining native stoichiometries of the protein of interest to the components of the cellular environment may be important for proteins that make specific interactions with cellular constituents.


Asunto(s)
Resonancia Magnética Nuclear Biomolecular , Protones , Resonancia Magnética Nuclear Biomolecular/métodos , Proteínas de Saccharomyces cerevisiae/química , Proteínas de Saccharomyces cerevisiae/análisis , Saccharomyces cerevisiae , Proteínas/química , Proteínas/análisis , Factores de Terminación de Péptidos/química
5.
Chem Rev ; 122(10): 10036-10086, 2022 05 25.
Artículo en Inglés | MEDLINE | ID: mdl-34878762

RESUMEN

Extracellular matrixes (ECMs), such as the cell walls and biofilms, are important for supporting cell integrity and function and regulating intercellular communication. These biomaterials are also of significant interest to the production of biofuels and the development of antimicrobial treatment. Solid-state nuclear magnetic resonance (ssNMR) and magic-angle spinning-dynamic nuclear polarization (MAS-DNP) are uniquely powerful for understanding the conformational structure, dynamical characteristics, and supramolecular assemblies of carbohydrates and other biomolecules in ECMs. This review highlights the recent high-resolution investigations of intact ECMs and native cells in many organisms spanning across plants, bacteria, fungi, and algae. We spotlight the structural principles identified in ECMs, discuss the current technical limitation and underexplored biochemical topics, and point out the promising opportunities enabled by the recent advances of the rapidly evolving ssNMR technology.


Asunto(s)
Pared Celular , Hongos , Bacterias , Pared Celular/química , Espectroscopía de Resonancia Magnética , Resonancia Magnética Nuclear Biomolecular , Plantas
6.
Phys Chem Chem Phys ; 26(6): 5669-5682, 2024 Feb 07.
Artículo en Inglés | MEDLINE | ID: mdl-38288878

RESUMEN

Two polarizing agents from the AsymPol family, AsymPol-TEK and cAsymPol-TEK (methyl-free version) are introduced for MAS-DNP applications in non-aqueous solvents. The performance of these new biradicals is rationalized in detail using a combination of electron paramagnetic resonance spectroscopy, density functional theory, molecular dynamics and quantitative MAS-DNP spin dynamics simulations. By slightly modifying the experimental protocol to keep the sample temperature low at insertion, we are able to obtain reproducable DNP-NMR data with 1,1,2,2-tetrachloroethane (TCE) at 100 K, which facilitates optimization and comparison of different polarizing agents. At intermediate magnetic fields, AsymPol-TEK and cAsymPol-TEK provide 1.5 to 3-fold improvement in sensitivity compared to TEKPol, one of the most widely used polarizing agents for organic solvents, with significantly shorter DNP build-up times of ∼1 s and ∼2 s at 9.4 and 14.1 T respectively. In the course of the work, we also isolated and characterized two diastereoisomers that can form during the synthesis of AsymPol-TEK; their difference in performance is described and discussed. Finally, the advantages of the AsymPol-TEKs are demonstrated by recording 2D 13C-13C correlation experiments at natural 13C-abundance of proton-dense microcrystals and by polarizing the surface of ZnO nanocrystals (NCs) coated with diphenyl phosphate ligands. For those experiments, cAsymPol-TEK yielded a three-fold increase in sensitivity compared to TEKPol, corresponding to a nine-fold time saving.

7.
J Am Chem Soc ; 145(38): 20749-20754, 2023 09 27.
Artículo en Inglés | MEDLINE | ID: mdl-37722679

RESUMEN

Nature is rich with examples of highly specialized biological materials produced by organisms for functions, including defense, hunting, and protection. Along these lines, velvet worms (Onychophora) expel a protein-based slime used for hunting and defense that upon shearing and dehydration forms fibers as stiff as thermoplastics. These fibers can dissolve back into their precursor proteins in water, after which they can be drawn into new fibers, providing biological inspiration to design recyclable materials. Elevated phosphorus content in velvet worm slime was previously observed and putatively ascribed to protein phosphorylation. Here, we show instead that phosphorus is primarily present as phosphonate moieties in the slime of distantly related velvet worm species. Using high-resolution nuclear magnetic resonance (NMR), natural abundance dynamic nuclear polarization (DNP), and mass spectrometry (MS), we demonstrate that 2-aminoethyl phosphonate (2-AEP) is associated with glycans linked to large slime proteins, while transcriptomic analyses confirm the expression of 2-AEP synthesizing enzymes in slime glands. The evolutionary conservation of this rare protein modification suggests an essential functional role of phosphonates in velvet worm slime and should stimulate further study of the function of this unusual chemical modification in nature.


Asunto(s)
Organofosfonatos , Proteínas , Proteínas/química , Espectroscopía de Resonancia Magnética , Fósforo , Espectrometría de Masas
8.
Faraday Discuss ; 241(0): 250-265, 2023 01 05.
Artículo en Inglés | MEDLINE | ID: mdl-36134444

RESUMEN

The possibility of enriching in 17O the water molecules within hydrated biominerals belonging to the Ca-pyrophosphate family was investigated, using liquid assisted grinding (LAG) in the presence of 17O-labelled water. Two phases with different hydration levels, namely triclinic calcium pyrophosphate dihydrate (Ca2P2O7·2H2O, denoted t-CPPD) and monoclinic calcium pyrophosphate tetrahydrate (Ca2P2O7·4H2O, denoted m-CPPT ß) were enriched in 17O using a "post-enrichment" strategy, in which the non-labelled precursors were ground under gentle milling conditions in the presence of stoichiometric quantities of 17O-enriched water (introduced here in very small volumes ∼10 µL). Using high-resolution 17O solid-state NMR (ssNMR) analyses at multiple magnetic fields, and dynamic nuclear polarisation (DNP)-enhanced 17O NMR, it was possible to show that the labelled water molecules are mainly located at the core of the crystal structures, but that they can enter the lattice in different ways, namely by dissolution/recrystallisation or by diffusion. Overall, this work sheds light on the importance of high-resolution 17O NMR to help decipher the different roles that water can play as a liquid-assisted grinding agent and as a reagent for 17O-isotopic enrichment.


Asunto(s)
Pirofosfato de Calcio , Difosfatos , Cristalización , Pirofosfato de Calcio/química , Agua/química
9.
J Am Chem Soc ; 144(51): 23448-23464, 2022 Dec 28.
Artículo en Inglés | MEDLINE | ID: mdl-36516873

RESUMEN

This work introduces an approach to uncoupling electrons via maximum utilization of localized aromatic units, i.e., the Clar's π-sextets. To illustrate the utility of this concept to the design of Kekulé diradicaloids, we have synthesized a tridecacyclic polyaromatic system where a gain of five Clar's sextets in the open-shell form overcomes electron pairing and leads to the emergence of a high degree of diradical character. According to unrestricted symmetry-broken UCAM-B3LYP calculations, the singlet diradical character in this core system is characterized by the y0 value of 0.98 (y0 = 0 for a closed-shell molecule, y0 = 1 for pure diradical). The efficiency of the new design strategy was evaluated by comparing the Kekulé system with an isomeric non-Kekulé diradical of identical size, i.e., a system where the radical centers cannot couple via resonance. The calculated singlet-triplet gap, i.e., the ΔEST values, in both of these systems approaches zero: -0.3 kcal/mol for the Kekulé and +0.2 kcal/mol for the non-Kekulé diradicaloids. The target isomeric Kekulé and non-Kekulé systems were assembled using a sequence of radical periannulations, cross-coupling, and C-H activation. The diradicals are kinetically stabilized by six tert-butyl substituents and (triisopropylsilyl)acetylene groups. Both molecules are NMR-inactive but electron paramagnetic resonance (EPR)-active at room temperature. Cyclic voltammetry revealed quasi-reversible oxidation and reduction processes, consistent with the presence of two nearly degenerate partially occupied molecular orbitals. The experimentally measured ΔEST value of -0.14 kcal/mol confirms that K is, indeed, a nearly perfect singlet diradical.

10.
Plant Cell ; 31(5): 1113-1126, 2019 05.
Artículo en Inglés | MEDLINE | ID: mdl-30886126

RESUMEN

Acetylation, a prevalent modification of cell-wall polymers, is a tightly controlled regulatory process that orchestrates plant growth and environmental adaptation. However, due to limited characterization of the enzymes involved, it is unclear how plants establish and dynamically regulate the acetylation pattern in response to growth requirements. In this study, we identified a rice (Oryza sativa) GDSL esterase that deacetylates the side chain of the major rice hemicellulose, arabinoxylan. Acetyl esterases involved in arabinoxylan modification were screened using enzymatic assays combined with mass spectrometry analysis. One candidate, DEACETYLASE ON ARABINOSYL SIDECHAIN OF XYLAN1 (DARX1), is specific for arabinosyl residues. Disruption of DARX1 via Tos17 insertion and CRISPR/Cas9 approaches resulted in the accumulation of acetates on the xylan arabinosyl side chains. Recombinant DARX1 abolished the excess acetyl groups on arabinoxylan-derived oligosaccharides of the darx1 mutants in vitro. Moreover, DARX1 is localized to the Golgi apparatus. Two-dimensional 13C-13C correlation spectroscopy and atomic force microscopy further revealed that the abnormal acetylation pattern observed in darx1 interrupts arabinoxylan conformation and cellulose microfibril orientation, resulting in compromised secondary wall patterning and reduced mechanical strength. This study provides insight into the mechanism controlling the acetylation pattern on arabinoxylan side chains and suggests a strategy to breed robust elite crops.


Asunto(s)
Oryza/enzimología , Proteínas de Plantas/metabolismo , Xilanos/metabolismo , Acetilación , Pared Celular/metabolismo , Pared Celular/ultraestructura , Celulosa/metabolismo , Productos Agrícolas , Esterasas/genética , Esterasas/metabolismo , Aparato de Golgi/metabolismo , Aparato de Golgi/ultraestructura , Mutación , Oligosacáridos/metabolismo , Oryza/genética , Oryza/ultraestructura , Fitomejoramiento , Proteínas de Plantas/genética
11.
Biomacromolecules ; 23(6): 2290-2301, 2022 06 13.
Artículo en Inglés | MEDLINE | ID: mdl-35341242

RESUMEN

Cellulose, the most abundant biopolymer, is a central source for renewable energy and functionalized materials. In vitro synthesis of cellulose microfibrils (CMFs) has become possible using purified cellulose synthase (CESA) isoforms from Physcomitrium patens and hybrid aspen. The exact nature of these in vitro fibrils remains unknown. Here, we characterize in vitro-synthesized fibers made by CESAs present in membrane fractions of P. patens over-expressing CESA5 by cryo-electron tomography and dynamic nuclear polarization (DNP) solid-state NMR. DNP enabled measuring two-dimensional 13C-13C correlation spectra without isotope-labeling of the fibers. Results show structural similarity between in vitro fibrils and native CMF in plant cell walls. Intensity quantifications agree with the 18-chain structural model for plant CMF and indicate limited fibrillar bundling. The in vitro system thus reveals insights into cell wall synthesis and may contribute to novel cellulosic materials. The integrated DNP and cryo-electron tomography methods are also applicable to structural studies of other carbohydrate-based biomaterials.


Asunto(s)
Bryopsida , Celulosa , Pared Celular/química , Celulosa/química , Tomografía con Microscopio Electrónico , Glucosiltransferasas/química , Espectroscopía de Resonancia Magnética/métodos
12.
J Phys Chem A ; 126(16): 2600-2608, 2022 Apr 28.
Artículo en Inglés | MEDLINE | ID: mdl-35417169

RESUMEN

We propose a new, more efficient, and potentially cost effective, solid-state nuclear spin hyperpolarization method combining the cross-effect mechanism and electron spin optical hyperpolarization in rotating solids. We first demonstrate optical hyperpolarization in the solid state at low temperatures and low field and then investigate its field dependence to obtain the optimal condition for high-field electron spin hyperpolarization. The results are then incorporated into advanced magic-angle spinning dynamic nuclear polarization (MAS-DNP) numerical simulations that show that optically pumped MAS-DNP could yield breakthrough enhancements at very high magnetic fields. Based on these investigations, enhancements greater than the ratio of electron to nucleus magnetic moments (>658 for 1H) are possible without microwave irradiation. This could solve at once the MAS-DNP performance decrease with increasing field and the high cost of MAS-DNP instruments at very high fields.

13.
Solid State Nucl Magn Reson ; 122: 101838, 2022 12.
Artículo en Inglés | MEDLINE | ID: mdl-36410100

RESUMEN

NMR is a valuable tool for studying insects. Solid-state NMR has been used to obtain the chemical composition and gain insight into the sclerotization process of exoskeletons. There is typically little difficulty in obtaining sufficient sample quantity for exoskeletons. However, obtaining enough sample of other insect components for solid-state NMR experiments can be problematic while isotopically enriching them is near impossible. This is especially the case for insect wing membranes which is of interest to us. Issues with obtaining sufficient sample are the thickness of wing membranes is on the order of microns, each membrane region is surrounded by veins and occupies a small area, and the membranes are separated from the wing by physical dissection. Accordingly, NMR signal enhancement methods are needed. MAS-DNP has a track record of providing significant signal enhancements for a wide variety of materials. Here we demonstrate that MAS-DNP is useful for providing high quality one-dimensional and two-dimensional solid-state NMR spectra on cicada wing membrane at natural isotopic abundance.


Asunto(s)
Insectos , Imagen por Resonancia Magnética , Animales , Examen Físico , Venas , Disección
14.
Angew Chem Int Ed Engl ; 61(12): e202114103, 2022 03 14.
Artículo en Inglés | MEDLINE | ID: mdl-35019217

RESUMEN

Efficiently hyperpolarizing proton-dense molecular solids through dynamic nuclear polarization (DNP) solid-state NMR is still an unmet challenge. Polarizing agents (PAs) developed so far do not perform well on proton-rich systems, such as organic microcrystals and biomolecular assemblies. Herein we introduce a new PA, cAsymPol-POK, and report outstanding hyperpolarization efficiency on 12.76 kDa U-13 C,15 N-labeled LecA protein and pharmaceutical drugs at high magnetic fields (up to 18.8 T) and fast magic angle spinning (MAS) frequencies (up to 40 kHz). The performance of cAsymPol-POK is rationalized by MAS-DNP simulations combined with electron paramagnetic resonance (EPR), density functional theory (DFT) and molecular dynamics (MD). This work shows that this new biradical is compatible with challenging biomolecular applications and unlocks the rapid acquisition of 13 C-13 C and 15 N-13 C correlations of pharmaceutical drugs at natural isotopic abundance, which are key experiments for structure determination.


Asunto(s)
Protones , Espectroscopía de Resonancia por Spin del Electrón , Espectroscopía de Resonancia Magnética , Preparaciones Farmacéuticas
15.
Anal Chem ; 93(46): 15365-15372, 2021 11 23.
Artículo en Inglés | MEDLINE | ID: mdl-34761922

RESUMEN

Biochar, a low-density yet carbon-rich material derived from different organic materials pyrolyzed under low or no oxygen conditions, has been widely studied as a soil amendment, for greenhouse gas mitigation and in remediation of trace element-contaminated soils. Molecular speciation of biochar compounds has been challenging due to low solubility, aggregation, and immense compositional polydispersity that challenges nearly all mass spectrometry methods routinely applied to carbon-based organic materials. Through a combined technique approach that applies advanced analytical strategies, we provide bulk and molecular characterization of Kentucky bluegrass biochar that can be applied to any biomass or biochar sample. First, we characterize Kentucky bluegrass biochar chemical functional groups by solid-state magic-angle spinning dynamic nuclear polarization NMR (MAS-DNP NMR) and resolve aromatic and aliphatic signals from the pyrogenic material and intact plant material. Next, we isolate water-soluble biochar species by solid-phase extraction followed by Fourier transform ion cyclotron resonance mass spectrometry (FT-ICR MS) and identify highly polar, oxygen species across a wide carbon number range. Solvent fractionation of biochar further expands the compositional range and identifies condensed polycyclic aromatic species across nonpolar and polar classes detected by two ionization modes (-ESI and +APPI) by FT-ICR MS. Plotting biochar species with DBE versus carbon number highlights the pericondensed molecular structural motif that persists across numerous heteroatom classes and ionization modes. To the best of our knowledge, this is the first molecular level identification of nonfunctionalized PAHs in biochar extracts by APPI FT-ICR MS. Thus, we identify biochar species that span the same compositional space as coal, heavy oil asphaltenes, and coal tar and correspond to condensed ring PAHs.


Asunto(s)
Carbón Orgánico , Agua , Espectrometría de Masas , Solventes
16.
Chemistry ; 27(49): 12574-12588, 2021 Sep 01.
Artículo en Inglés | MEDLINE | ID: mdl-34131984

RESUMEN

In recent years, there has been increasing interest in developing cost-efficient, fast, and user-friendly 17 O enrichment protocols to help to understand the structure and reactivity of materials by using 17 O NMR spectroscopy. Here, we show for the first time how ball milling (BM) can be used to selectively and efficiently enrich the surface of fumed silica, which is widely used at industrial scale. Short milling times (up to 15 min) allowed modulation of the enrichment level (up to ca. 5 %) without significantly changing the nature of the material. High-precision 17 O compositions were measured at different milling times by using large-geometry secondary-ion mass spectrometry (LG-SIMS). High-resolution 17 O NMR analyses (including at 35.2 T) allowed clear identification of the signals from siloxane (Si-O-Si) and silanols (Si-OH), while DNP analyses, performed by using direct 17 O polarization and indirect 17 O{1 H} CP excitation, agreed with selective labeling of the surface. Information on the distribution of Si-OH environments at the surface was obtained from 2D 1 H-17 O D-HMQC correlations. Finally, the surface-labeled silica was reacted with titania and using 17 O DNP, their common interface was probed and Si-O-Ti bonds identified.

17.
Phys Chem Chem Phys ; 23(24): 13768-13769, 2021 Jun 23.
Artículo en Inglés | MEDLINE | ID: mdl-34115087

RESUMEN

Correction for 'De novo prediction of cross-effect efficiency for magic angle spinning dynamic nuclear polarization' by Frédéric Mentink-Vigier et al., Phys. Chem. Chem. Phys., 2019, 21, 2166-2176, DOI: 10.1039/C8CP06819D.

18.
Magn Reson Chem ; 59(9-10): 991-1008, 2021 Sep.
Artículo en Inglés | MEDLINE | ID: mdl-33624858

RESUMEN

Detecting proximities between nuclei is crucial for atomic-scale structure determination with nuclear magnetic resonance (NMR) spectroscopy. Different from spin-1/2 nuclei, the methodology for quadrupolar nuclei is limited for solids due to the complex spin dynamics under simultaneous magic-angle spinning (MAS) and radio-frequency irradiation. Herein, the performances of several homonuclear rotary recoupling (HORROR)-based homonuclear dipolar recoupling sequences are evaluated for 27 Al (spin-5/2). It is shown numerically and experimentally on mesoporous alumina that BR 2 2 1 outperforms the supercycled S3 sequence and its pure double-quantum (DQ) (bracketed) version, [S3 ], both in terms of DQ transfer efficiency and bandwidth. This result is surprising since the S3 sequence is among the best low-power recoupling schemes for spin-1/2. The superiority of BR 2 2 1 is thoroughly explained, and the crucial role of radio-frequency offsets during its spin dynamics is highlighted. The analytical approximation of BR 2 2 1 , derived in an offset-toggling frame, clarifies the interplay between offset and DQ efficiency, namely, the benefits of off-resonance irradiation and the trough in DQ efficiency for BR 2 2 1 when the irradiation is central between two resonances, both for spin-1/2 and half-integer-spin quadrupolar nuclei. Additionally, density matrix propagations show that the BR 2 2 1 sequence, applied to quadrupolar nuclei subject to quadrupolar interaction much larger than radio-frequency frequency field, can create single- and multiple-quantum coherences for near on-resonance irradiation. This significantly perturbs the creation of DQ coherences between central transitions of neighboring quadrupolar nuclei. This effect explains the DQ efficiency trough for near on-resonance irradiation, in the case of both cross-correlation and autocorrelation peaks. Overall, this work aids experimental acquisition of homonuclear dipolar correlation spectra of half-integer-spin quadrupolar nuclei and provides theoretical insights towards improving recoupling schemes at high magnetic field and fast MAS.

19.
Biochem Soc Trans ; 48(3): 1089-1099, 2020 06 30.
Artículo en Inglés | MEDLINE | ID: mdl-32379300

RESUMEN

Solid-state nuclear magnetic resonance (ssNMR) is an indispensable tool for elucidating the structure and dynamics of insoluble and non-crystalline biomolecules. The recent advances in the sensitivity-enhancing technique magic-angle spinning dynamic nuclear polarization (MAS-DNP) have substantially expanded the territory of ssNMR investigations and enabled the detection of polymer interfaces in a cellular environment. This article highlights the emerging MAS-DNP approaches and their applications to the analysis of biomolecular composites and intact cells to determine the folding pathway and ligand binding of proteins, the structural polymorphism of low-populated biopolymers, as well as the physical interactions between carbohydrates, proteins, and lignin. These structural features provide an atomic-level understanding of many cellular processes, promoting the development of better biomaterials and inhibitors. It is anticipated that the capabilities of MAS-DNP in biomolecular and biomaterial research will be further enlarged by the rapid development of instrumentation and methodology.


Asunto(s)
Proteínas de la Membrana/química , Resonancia Magnética Nuclear Biomolecular , Bacterias , Biopolímeros , Membrana Celular/química , Núcleo Celular , Pared Celular/química , Hongos , Humanos , Ligandos , Plantas , Polisacáridos/química , Unión Proteica , Temperatura
20.
Inorg Chem ; 59(18): 13050-13066, 2020 Sep 21.
Artículo en Inglés | MEDLINE | ID: mdl-32167301

RESUMEN

While 17O NMR is increasingly being used for elucidating the structure and reactivity of complex molecular and materials systems, much effort is still required for it to become a routine analytical technique. One of the main difficulties for its development comes from the very low natural abundance of 17O (0.04%), which implies that isotopic labeling is generally needed prior to NMR analyses. However, 17O-enrichment protocols are often unattractive in terms of cost, safety, and/or practicality, even for compounds as simple as metal oxides. Here, we demonstrate how mechanochemistry can be used in a highly efficient way for the direct 17O isotopic labeling of a variety of s-, p-, and d-block oxides, which are of major interest for the preparation of functional ceramics and glasses: Li2O, CaO, Al2O3, SiO2, TiO2, and ZrO2. For each oxide, the enrichment step was performed under ambient conditions in less than 1 h and at low cost, which makes these synthetic approaches highly appealing in comparison to the existing literature. Using high-resolution solid-state 17O NMR and dynamic nuclear polarization, atomic-level insight into the enrichment process is achieved, especially for titania and alumina. Indeed, it was possible to demonstrate that enriched oxygen sites are present not only at the surface but also within the oxide particles. Moreover, information on the actual reactions occurring during the milling step could be obtained by 17O NMR, in terms of both their kinetics and the nature of the reactive species. Finally, it was demonstrated how high-resolution 17O NMR can be used for studying the reactivity at the interfaces between different oxide particles during ball-milling, especially in cases when X-ray diffraction techniques are uninformative. More generally, such investigations will be useful not only for producing 17O-enriched precursors efficiently but also for understanding better mechanisms of mechanochemical processes themselves.

SELECCIÓN DE REFERENCIAS
DETALLE DE LA BÚSQUEDA