Pulsed Field Gradient Diffusion Research Articles

A Complementary Multitechnique Approach to Assess the Bias in Molecular Weight Determination of Lignin by Derivatization-Free Gel Permeation Chromatography.

The growing interest in lignin valorization in the past decades calls for analytical techniques for lignin characterization, ranging from wet chemistry techniques to highly sophisticated chromatographic and spectroscopic methods. One of the key parameters to consider is the molecular weight profile of lignin, which is routinely determined by size-exclusion chromatography; however, this is by no means straightforward and is prone to being hampered by considerable errors. Our study expands the fundamental understanding of the bias-inducing mechanisms in gel permeation chromatography (GPC), the magnitude of error originating from using polystyrene standards for mass calibration, and an evaluation of the effects of the solvent and type of lignin on the observed bias. The developed partial least-squares (PLS) regression model for lignin-related monomers revealed that lignin is prone to association mainly via hydrogen bonding. This hypothesis was supported by functional group-based analysis of the bias as well as pulse field gradient (pfg) diffusion NMR spectroscopy of model compounds in THF-d8. Furthermore, although the lack of standards hindered drawing conclusions based on functionalities, direct infusion electrospray ionization mass spectrometry indicated that the relative bias decreases considerably for higher molecular weight species. The results from pfg-diffusion NMR spectroscopy on whole lignin samples were comparable when the same solvents were used in both experiments; in addition, the comparison between results obtained by pfg-diffusion NMR in different solvents gives some additional insights into the aggregation.

Insight into the molecular basis of the soluble storage-state to fibrous-state structural transformation of spider pyriform silk

Pyriform (or piriform) silk is a fibrous spider silk used in a composite material to attach web silks to substrates and disparate silk types together. Argiope argentata pyriform silk is formed from a protein with a central repetitive region consisting of a 234-amino acid unit (the "Py unit") repeated 21 times. We have previously shown that the Py unit behaves modularly, with a structured core of 6 α-helices that retains its structuring upon truncation of N- and C-terminal intrinsically disordered segments. Here, we apply solution-state nuclear magnetic resonance (NMR) spectroscopy to investigate the structured core of the Py unit. Based on 15N spin relaxation measurements and reduced spectral density mapping at two field strengths, the α-helical core of the Py unit exhibits minimal backbone motion beyond the tumbling of the globular domain except for one interhelical linker that exhibits a notably higher degree of dynamics. Evaluation of rotational diffusion based on 15N spin relaxation and translational diffusion by pulsed field gradient diffusion NMR demonstrates that the core of the Py unit behaves as a compact structure, with hydrodynamic behaviour consistent with a well-packed protein. Pyriform silk protein undergoes a structural transformation upon conversion from the soluble-state to the fibrous-state, with a loss of αhelical content and gain of β-sheet content. The more dynamic segment we observe midway through the structured core of the Py core, especially in the context of the intrinsically disordered segments that flank the core, would provide an initiation point for decompaction, structural transformation, and/or intermolecular interactions.

Lithium Ion Battery Electrolytes Based on Deep Eutectic Solvents

Deep eutectic solvents (DESs) have emerged as an alternative to both common organic solvents and ionic liquids (ILs). DESs share physicochemical properties with ILs such as low vapor pressure, high thermal stability, while offering advantages such as low toxicity, lower cost, and ease of preparation. Moreover, DESs are attractive candidates for electrochemical applications due to their large voltage windows and solubility properties. DESs as a solvent class share a general composition of a hydrogen bond donor (HBD), typically a polyol, amide, or acid, and a hydrogen bond acceptor (HBA), usually a quaternary ammonium or phosphonium salt. At a specific molar composition of a HBD and HBA, the DES forms a eutectic mixture resulting in a large melting point depression.Despite being widely studied, the microscopic structures of DESs have remained largely uncharacterized. Herein, we present a multitechnique NMR study of ethaline (ethylene glycol + choline chloride). Pulsed field gradient diffusion, nuclear Overhauser effect spectroscopy (NOESY), 13C NMR relaxation experiments are deployed to sample a range of frequencies and modes of motion of the polyol and choline components of the DES. Generally, translational and rotational diffusion of polyols are more rapid than those of choline while short-range rotational motions observed from 13C relaxation indicate slow local motion of ethylene glycol at low choline chloride (ChCl) content.Additionally, we have investigated the solubility properties of several lithium salts (LiTFSI, LiFSI, LiBF4, and LiOAc) in these DESs and utilized the same suite of NMR techniques to understand how they act as solvents. One interesting observation is that the Li+ diffusivity exceeds that of the counterion for TFSI-, but not for FSI-. Finally, large changes in the structural organization of DESs that result from the presence of lithium salts may serve as a guide to the design of a new class of electrolytes for lithium-ion batteries.

Double pulsed field gradient diffusion MRI to assess skeletal muscle microstructure.

Preliminary study to determine whether double pulsed field gradient (PFG) diffusion MRI is sensitive to key features of muscle microstructure related to function. The restricted diffusion profile of molecules in models of muscle microstructure derived from histology were systematically simulated using a numerical simulation approach. Diffusion tensor subspace imaging analysis of the diffusion signal was performed, and spherical anisotropy (SA) was calculated for each model. Linear regression was used to determine the predictive capacity of SA on the fiber area, fiber diameter, and surface area to volume ratio of the models. Additionally, a rat model of muscle hypertrophy was scanned using a single PFG and a double PFG pulse sequence, and the restricted diffusion measurements were compared with histological measurements of microstructure. Excellent agreement between SA and muscle fiber area (r2 = 0.71; p < 0.0001), fiber diameter (r2 = 0.83; p < 0.0001), and surface area to volume ratio (r2 = 0.97; p < 0.0001) in simulated models was found. In a scanned rat leg, the distribution of these microstructural features measured from histology was broad and demonstrated that there is a wide variance in the microstructural features observed, similar to the SA distributions. However, the distribution of fractional anisotropy measurements in the same tissue was narrow. This study demonstrates that SA-a scalar value from diffusion tensor subspace imaging analysis-is highly sensitive to muscle microstructural features predictive of function. Furthermore, these techniques and analysis tools can be translated to real experiments in skeletal muscle. The increased dynamic range of SA compared with fractional anisotropy in the same tissue suggests increased sensitivity to detecting changes in tissue microstructure.

Silicone-specific identification of trace polydimethylsiloxanes in wines with 2D-diffusion-ordered nuclear magnetic resonance spectroscopy (DOSY-NMR)

Present work stresses a novel analytical approach for increasing the specificity of standard NMR approaches for identifying polydimethylsiloxane (PDMS) and further silicone moieties in wines’ organic extracts, by including a second dimension that correlates chemical shifts with diffusion coefficients by means of pulsed-field gradient diffusion ordered spectroscopy (DOSY-NMR). Each silicone source in wines is unambiguously assigned by correlation of both local chemical environments and by a unique diffusion coefficient value, in turn related to a hydrodynamic radius (RH) that can be obtained with respect proper internal standards. Obtained PDMS diffusion coefficient values and hydrodynamic radii in wines’ extracts, in agreement with expected values, present a selectivity and specificity so far not reported, that positions DOSY-NMR spectroscopy as an alternative in oenology for controlling PDMS limits.

(Digital Presentation) Fundamentals of Deep Eutectic Solvents As Electrolytes for Lithium-Ion Batteries

Deep eutectic solvents (DESs) have emerged as an alternative to both common organic solvents and ionic liquids (ILs). DESs share physicochemical properties with ILs such as low vapor pressure, high thermal stability, high viscosity while offering advantages such as low toxicity, lower cost, and ease of preparation. Moreover, DESs are attractive candidates for electrochemical applications due to their large voltage windows and solubility properties. DESs as a solvent class share a general composition of a hydrogen bond donor (HBD), typically a polyol, amide, or acid, and a hydrogen box acceptor (HBA), usually a quaternary ammonium or phosphonium salt. At a specific molar composition of a HBD and HBD, the DES forms a eutectic mixture resulting in a large melting point depression due to extensive hydrogen bonding between the components.Despite being widely studied, the microscopic structures of DESs have remained largely uncharacterized. Herein, we present a multitechnique NMR study of two DESs: glyceline (glycerol + choline chloride) and ethaline (ethylene glycol + choline chloride). Fast-field cycling 1H relaxometry, pulsed field gradient diffusion, nuclear Overhauser effect spectroscopy (NOESY), 13C NMR relaxation, and pressure-dependent NMR experiments are deployed to sample a range of frequencies and modes of motion of the polyol and choline components of the DES. Generally, translational and rotational diffusion of polyols are more rapid than those of choline while short-range rotational motions observed from 13C relaxation indicate slow local motion of glycerol at low choline chloride (ChCl) content. We show how the additional hydroxyl group present in glycerol contributes to not only higher viscosities, but a larger perturbation of the hydrogen bonding network by the addition of ChCl.Additionally, we have investigated the solubility properties of several lithium salts (LiTFSI, LiFSI, LiPF6, LiBF4, and LiOAc) in these DESs and utilized the same suite of NMR techniques to understand how they act as solvents. We observe that due to anion effects, the heterogeneities present in DES result in differential solvation for some species where there is a distinction of lithium salts co-existing in these holes as well as the bulk. The large changes in the structural organization of DESs that result from the presence of lithium salts will serve as a guide to the design of a new class of electrolytes for lithium-ion batteries.

(Invited) NMR Investigation of Proton Transport in Concentrated Aqueous and Polymer Electrolytes Based on Polyphosphoric Acid

Concentrated aqueous electrolytes have received much attention in recent years due to their high ionic conductivity and electrochemical window that usually exceeds that of the water electrolysis limitation. In collaboration with Ziyue Li and Fei Wang (Fudan University) we have investigated water/polyphosphoric acid solutions ranging in concentration from 1.25 to 16.6 molar being considered for proton-based batteries. Self-diffusion coefficients measured by pulsed field gradient NMR as well as spin-lattice time relaxation measurements were determined for 31P and 1H. The results shed light on the observed maximum in conductivity at the 5.9 molar concentration.For polymer electrolyte membrane fuel cells (PEMFC), polybenzimidazole (PBI) membranes with high phosphoric acid content have been developed previously by collaborator Brian Benicewicz (University of South Carolina) using the so-called PPA process. Recently, a carefully controlled drying method has been discovered by Benicewicz and co-worker Laura Murdock in which a PBI membrane prepared by the PPA process is transformed into a dense PBI film. Though originally developed for flow battery applications, it was found that this dense film could be re-acidified to yield a mechanically robust PEM. Furthermore this method provides a synthetic route to PBI films without the use of any organic solvents. The re-doped PBI films display high ionic conductivity at elevated temperatures, similar to the starting PBI gel membranes prepared by the PPA process, while containing nearly a factor of two less acid than the latter and also exhibiting enhanced mechanical properties. NMR spectroscopy was used to characterize the structure and dynamics of the modified and the original PBI/PPA membranes. In addition to pulsed field gradient diffusion studies, we have performed solid state one-dimensional 31P, 13C and two-dimensional 31P{1H} and 13C{1H} HETCOR NMR measurements, which yield additional information on structural differences between the membranes and how these differences may affect transport properties.

Dynamics of Glyceline and Interactions of Constituents: A Multitechnique NMR Study.

The dynamics of the organic components of the deep eutectic solvent (DES) glyceline are analyzed using an array of complementary nuclear magnetic resonance (NMR) methods. Fast-field cycling 1H relaxometry, pulsed field gradient diffusion, nuclear overhauser effect spectroscopy (NOESY), 13C NMR relaxation, and pressure-dependent NMR experiments are deployed to sample a range of frequencies and modes of motion of the glycerol and choline components of the DES. Generally, translational and rotational diffusion of glycerol are more rapid than those of choline while short-range rotational motions observed from 13C relaxation indicate slow local motion of glycerol at low choline chloride (ChCl) content. The rates of glycerol and choline local motions become more similar at higher ChCl. This result taken together with pressure-dependent NMR studies show that the addition of ChCl makes it easier to disrupt glycerol packing. Finally, a relatively slow hydroxyl H-exchange process between glycerol and choline protons is deduced from the data. Consistent with this, NOESY results indicate relatively little direct H-bonding between glycerol and choline. These results suggest that the glycerol H-bonding network is disrupted as choline is added, but primarily in regions where there is intimate mixing of the two components. Thus, the local dynamics of most of the glycerol resembles that of pure glycerol until substantial choline chloride is present.

Extremely High Proton Mobility in Nanoscopic Titania Hydrates

Using protons as the ionic charge carriers in energy conversion and storage devices is an attractive prospect, but designing electrode/electrolyte materials with fast proton mobility at room temperature is still a challenge. Here we report on two novel titania hydrates with high proton mobility: one is one-dimensional H2Ti5O11·H2O material with ~2 µm length and ~150 nm width; the other is zero-dimensional TiO2·0.4H2O material with average dimension ~5 nm. To gain a deeper understanding of the correlation between materials local structure and fast proton transport mechanisms, extended X-ray absorption fine structure, high resolution transmission electron microscopy, and nuclear magnetic resonance pulsed field gradient diffusion measurements were conducted. The H-bonded species are identified with nano-confined water located between the TiO6 octahedra layered structures in H2Ti5O11·H2O and on the surface of TiO6 octahedra in TiO2·0.4H2O. The proton self-diffusion coefficients at room temperature are about 4×10−9 m2 s−1 for both H2Ti5O11·H2O and TiO2·0.4H2O, which exceed that in liquid water at ambient temperature by nearly a factor of two. These values decrease to about 1×10−9 m2 s−1 at −25 oC in both materials. These unusually high diffusion coefficients imply a Grotthuss transport mechanism and suggest attractive possible application in proton batteries or resistive switching. Further, this materials protocol can create new opportunities in designing other proton-based electrodes and electrolytes with nano-confined water as the fast proton carrier in solids.

NMR investigation of the thermogelling properties, anomalous diffusion, and structural changes in a Pluronic F127 triblock copolymer in the presence of gold nanoparticles

We studied the thermogelation of a triblock copolymer Pluronic F127 (poly(ethylene oxide)-poly(propylene oxide)-poly(ethylene oxide)) in an aqueous solvent in the presence of gold nanoparticles, using pulsed field gradient diffusion, NMR temperature experiments, relaxation measurements, and 2D heteronuclear NMR experiments. Pulsed field gradient diffusion NMR is a powerful technique to study the transition between diffusive regimes in a polymer mesh which are modulated by phase transitions in the polymeric network. In the isotropic phase, the triblock copolymer diffusion is a classical Fickian process. As the onset of gelation occurs, diffusion in the system becomes anomalous and the mean square displacement in the direction of the applied magnetic field gradient shows a power law dependence. Our experiments show that the introduction of gold nanoparticles leads to a disruption of gelation and the shifting of the formation of the ordered phase of the triblock copolymer to a higher temperature.

NMR Relaxometry and Diffusometry Analysis of Dynamics in Ionic Liquids and Ionogels for Use in Lithium-Ion Batteries.

We have investigated the charge transport dynamics of a novel solid-like electrolyte material based on mixtures of the ionic liquid (IL) 1-butyl-3-methylimidazolium bis(trifluoromethylsulfonyl)imide ([BMIM] TFSI) and various concentrations of lithium salt bis(trifluoromethylsulfonyl)imide (LiTFSI) confined within a SiO2 matrix, prepared via a sol-gel method. The translational diffusion coefficients of BMIM+, TFSI-, and Li+ in ILs and confined ILs (ionogels, IGs) with different concentrations of lithium salt have been measured at variable temperatures, covering the 20-100 °C range, using nuclear magnetic resonance (NMR) pulsed field gradient diffusion spectroscopy. The mobility of BMIM+, TFSI-, and Li+ was found to increase with the [BMIM] TFSI/LiTFSI ratio, exhibiting an almost liquid-like mobility in IGs. Additionally, the effect of confinement on IL rotational dynamics has been analyzed by measuring 1H, 19F, and 7Li spin-lattice relaxation rate dispersions of IGs at different temperatures, using fast field-cycling NMR relaxometry. The analysis of the experimental data was performed assuming the existence of two fractions of the liquid: a bulk fraction (at least several ionic radii from the silica particles) and a surface fraction (close to the silica particles) and using two different models based on translational and rotational diffusion and reorientation mediated by translational displacements. The existence and weighting of these two fractions of ions were obtained from the direct diffusion measurements. The results show that the ion dynamics slowed only modestly under confinement, which evidences that IGs preserve IL transport properties, and this behavior is an encouraging indication for using IGs as a solid electrolyte for Li+ batteries.

Protein structural changes characterized by high-pressure, pulsed field gradient diffusion NMR spectroscopy

Pulsed-field gradient NMR spectroscopy is widely used to measure the translational diffusion and hydrodynamic radius (Rh) of biomolecules in solution. For unfolded proteins, the Rh provides a sensitive reporter on the ensemble-averaged conformation and the extent of polypeptide chain expansion as a function of added denaturant. Hydrostatic pressure is a convenient and reversible alternative to chemical denaturants for the study of protein folding, and enables NMR measurements to be performed on a single sample. While the impact of pressure on the viscosity of water is well known, and our water diffusivity measurements agree closely with theoretical expectations, we find that elevated pressures increase the Rh of dioxane and other small molecules by amounts that correlate with their hydrophobicity, with parallel increases in rotational friction indicated by 13C longitudinal relaxation times. These data point to a tighter coupling with water for hydrophobic surfaces at elevated pressures. Translational diffusion measurement of the unfolded state of a pressure-sensitized ubiquitin mutant (VA2-ubiquitin) as a function of hydrostatic pressure or urea concentration shows that Rh values of both the folded and the unfolded states remain nearly invariant. At ca 23 Å, the Rh of the fully pressure-denatured state is essentially indistinguishable from the urea-denatured state, and close to the value expected for an idealized random coil of 76 residues. The intrinsically disordered protein (IDP) α-synuclein shows slight compaction at pressures above 2 kbar. Diffusion of unfolded ubiquitin and α-synuclein is significantly impacted by sample concentration, indicating that quantitative measurements need to be carried out under dilute conditions.

Magnetic resonance diffusion measurements of droplet size in drilling fluid emulsions on a benchtop instrument

Oil-based drilling fluids are complex emulsio-dispersions used in the construction of subsurface wellbores. Design and control of the rheological properties are crucial for successful drilling operations because the fluid performs several critical functions, such as hole cleaning (particle transport) and ensuring wellbore stability. One of the physical characteristics of the fluid that relates to rheology is the size of the water-in-oil emulsion droplets. Here we demonstrate the use of nuclear magnetic resonance (NMR) pulsed field gradient (PFG) diffusion measurements to determine droplet size distributions in oil-based drilling fluid analogues and a commercial formulation. NMR PFG allows these optically opaque samples to be measured without dilution, and the measurement is sensitive only to the liquid components. Although the NMR technique is established, the application to drilling fluids is novel. Through simulation and experiment, the suitability of a low-field benchtop NMR instrument (commonly used for rock core analysis in the petroleum industry) for measuring droplet size distributions in these fluids is explored. Despite limitations on the available gradient strength and the bulk diffusion coefficient of the brine, droplet radii in the range a = 1 μm to 50 μm are measured. The low-field NMR instrument is shown to be suitable for characterizing drilling fluids and similar complex emulsions.

Apelin conformational and binding equilibria upon micelle interaction primarily depend on membrane-mimetic headgroup

Apelin is one of two peptide hormones that activate the apelin receptor (AR or APJ) to regulate the cardiovascular system, central nervous system, and adipoinsular axis. Here, we apply circular dichroism (CD) spectropolarimetry and nuclear magnetic resonance (NMR) spectroscopy to characterize the potential membrane binding by the two longest bioactive apelin isoforms, apelin-55 and -36, using membrane-mimetic dodecylphosphocholine (DPC), sodium dodecyl sulfate (SDS), and 1-palmitoyl-2-hydroxy-sn-glycero-3-[phospho-rac-(1-glycerol)] (LPPG) micelles. Pulsed field gradient diffusion NMR experiments demonstrated preferential interaction of both apelin-55 and -36 with anionic SDS and LPPG micelles over zwitterionic DPC micelles. Chemical shift perturbations and changes in ps-ns scale dynamics of apelin-55 in all micelles were similarly localized along the polypeptide backbone, demonstrating clear dependence upon detergent headgroup, while comparison of chemical shifts between apelin-55 and apelin-36 showed negligible differences indicative of highly similar modes of micelle interaction. Notably, the observed behaviour was consistent with an ensemble averaged pair of free and bound states in fast exchange on the NMR timescale proportional to the fraction of micelle-bound protein, implying a similar conformational equilibrium regardless of headgroup and tailgroup. Membrane catalysis of apelin-AR binding would thus give rise to analogous behaviour in the essential C-terminal region common to all apelin isoforms.

Cation Dynamics in Supercooled and Solid Alkyl Methylimidazolium Bromide Ionic Liquids

The molecular dynamics of alkyl methylimidazolium bromide ionic liquids with different side groups of the cation are studied over a wide range of temperatures, covering the supercooled and crystalline states. Nuclear magnetic resonance relaxation dispersion (NMRD) at different magnetic field strengths was combined with NMR pulsed field gradient (PFG) diffusion measurements in order to obtain a description of the temperature dependence of the cationic mobility. While an Arrhenius dependence of the correlation times was found at high temperatures, a deviation is observed below a critical temperature of Tdyn ∼ 275 K which corresponds to about 1.25 Tg for two of the substances. The macroscopic diffusion coefficient, on the other hand, is best described by a VFT dependence down to a similar temperature, and a much weaker temperature dependence below. Measurements carried out in the crystalline state of 1-butyl-3-methylimidazolium bromide (Bmim Br) exhibit a dramatically increased self-diffusion coefficient in agreement with earlier reports of strong dynamic heterogeneity in the presence of minute amounts of water.

(Invited) Liquid and Solid State NMR Investigations of Electrolytes for Beyond Lithium Ion Applications

Nuclear magnetic resonance (NMR) has been productively employed to investigate ion transport and solvation in standard carbonate-based Li ion battery electrolytes and in solid polymer electrolytes based on poly(ethylene oxide) (PEO). Future electrochemical power sources require new electrolytes to adapt to disruptive changes in the basic working chemistry, such as moving from Li ion to Na ion, or to Li metal electrodes. We highlight two recent collaborative activities on electrolytes in our group (i) Na ion; (ii) Li metal polymer. Using a combination of 23Na, 19F and natural abundance 17O NMR, and pulsed field gradient diffusion in a collaboration with the U.S. Army Research Lab, we have examined NaPF6 solutions in binary solvent mixtures ethylene carbonate (EC) and ethylmethyl carbonate (EMC) for possible cation solvation preference as a function of EC/EMC ratio. Spectroscopic evidence demonstrate that the Li+ and Na+ cations share a number of similar ion–solvent and ion-ion interaction trends, such as a preference for a solvation shell rich in cyclic carbonates over linear carbonates and fluorinated carbonates. However some differences are noted, in particular a somewhat weaker ion-solvent interaction for Na+. Ionic Materials, Inc. has invented a novel polymer with extremely high ionic conductivity over a range of temperatures, even surpassing that of a commercial porous separator containing the standard liquid carbonate-based electrolyte at room temperature. This solid polymer can be reliably extruded into very thin films, is non-flammable, has attractive mechanical properties for lithium dendrite suppression, is electrochemically stable against Li, and is compatible with a variety of different anodes and cathodes. The polymer electrolyte is based on an inexpensive semicrystalline commercially available polymer such as polyether ether ketone or polyphenylene sulfide and Li salts familiar to the battery and polymer electrolyte communities. The ionic transport mechanism is unlike that which characterizes the PEO salt complexes – that is ion mobility is completely decoupled from polymer host motion, as verified by differential scanning calorimetry and NMR. In fact NMR pulsed gradient measurements reveal Li self-diffusion coefficients at room temperature that are an order of magnitude higher than in the ceramic ion conductors of the Li10GeP2S12 class and thus the highest in any known solid. Furthermore, the Li+ transference number determined electrochemically and by NMR is > 0.6.

Local diffusion and diffusion-T2 distribution measurements in porous media

Slice-selective pulsed field gradient (PFG) and PFG-T2 measurements are developed to measure spatially-resolved molecular diffusion and diffusion-T2 distributions. A spatially selective adiabatic inversion pulse was employed for slice-selection. The slice-selective pulse is able to select a coarse slice, on the order of 1cm, at an arbitrary position in the sample.The new method can be employed to characterize oil-water mixtures in porous media. The new technique has an inherent sensitivity advantage over phase encoding imaging based methods due to signal being localized from a thick slice. The method will be advantageous for magnetic resonance of porous media at low field where sensitivity is problematic.Experimental CPMG data, following PFG diffusion measurement, were compromised by a transient ΔB0(t) field offset. The off resonance effects of ΔB0(t) were examined by simulation. The ΔB0 offset artifact in D-T2 distribution measurements may be avoided by employing real data, instead of magnitude data.

Apela exhibits isoform- and headgroup-dependent modulation of micelle binding, peptide conformation and dynamics

Apela (also referred to as ELABELA and toddler) is a peptide hormone that activates the apelin receptor (AR or APJ) to regulate cardiovascular system development and function. Here, we report the first biophysical characterization of three apela isoforms, apela-54, -32, and -11, alongside a monomeric C1S-apela-11 mutant, using circular dichroism (CD) spectropolarimetry and nuclear magnetic resonance (NMR) spectroscopy. The behaviour of apela-54 is consistent with a preprotein containing a hydrophobic, N-terminal signal peptide. The potential for apela-membrane binding, leading to membrane catalyzed interactions with AR, was tested comprehensively for apela-32 and -11 in the presence of membrane-mimetic dodecylphosphocholine (DPC), sodium dodecyl sulfate (SDS), and 1-palmitoyl-2-hydroxy-sn-glycero-3-[phospho-rac-(1-glycerol)] (LPPG) micelles. According to pulsed-field gradient diffusion NMR experiments, apela-32 interacts with all three micelles. Chemical shift perturbations indicate widespread interactions along apela, with DPC and LPPG micelles inducing short segments with α-helical character at distinct regions. Consistent with these data, ps-ns dynamics along the peptide backbone appear decreased in the presence of micelles. Apela-11 and C1S-apela-11, alternatively, interact preferentially with SDS and LPPG micelles, promoting β-turn character observable by CD. Distinct differences in membrane-interaction propensity are therefore apparent both as a function of apela isoform and of detergent headgroup. These results imply the potential for cell membrane involvement in apela-AR recognition and binding, with the implication that membrane catalysis has distinct functional and regulatory roles throughout the apelinergic system.

Differentiating brown and white adipose tissues by high-resolution diffusion NMR spectroscopy

There are two types of fat tissues, white adipose tissue (WAT) and brown adipose tissue (BAT), which essentially perform opposite functions in whole body energy metabolism. There is a large interest in identifying novel biophysical properties of WAT and BAT by a quantitative and easy-to-run technique. In this work, we used high-resolution pulsed field gradient diffusion NMR spectroscopy to study the apparent diffusion coefficient (ADC) of fat molecules in rat BAT and WAT samples. The ADC of fat in BAT and WAT from rats fed with a chow diet was compared with that of rats fed with a high-fat diet to monitor how the diffusion properties change due to obesity-associated parameters such as lipid droplet size, fatty acid chain length, and saturation. Feeding a high-fat diet resulted in increased saturation, increased chain lengths, and reduced ADC of fat in WAT. The ADC of fat was lower in BAT relative to WAT in rats fed both chow and high-fat diets. Diffusion of fat was restricted in BAT due to the presence of small multilocular lipid droplets. Our findings indicate that in vivo diffusion might be a potential way for better delineation of BAT and WAT in both lean and obese states.

(Invited) NMR Investigation of Ion Transport and Solvation in Electrolytes for Beyond Lithium Ion Applications

Nuclear magnetic resonance (NMR) has been productively employed to investigate ion transport and solvation in standard carbonate-based Li ion battery electrolytes. Future electrochemical power sources require new electrolytes to adapt to disruptive changes in the basic working chemistry, such as moving from Li to Na. This work extends the application of some of the NMR tools developed by our lab and others to study the standard electrolytes to the new electrolytes. Using a combination of 23Na, 19F and natural abundance 17O NMR, and pulsed field gradient diffusion, we have examined NaPF6 solutions in binary solvent mixtures ethylene carbonate (EC) and ethylmethyl carbonate (EMC) for possible cation solvation preference as a function of EC/EMC ratio. Using similar methods, we also investigate the transport properties of newly discovered highly conductive ultra-concentrated aqueous solutions of LiTFSI.