Noncanonical Relationship between Heterogeneity and the Stokes-Einstein Breakdown in Deep Eutectic Solvents.

The relationship between Stokes-Einstein breakdown (SEB) and dynamical heterogeneity (DH) is of paramount importance in the physical chemistry of complex fluids. In this work, we employ neutron scattering to probe the DH and SEB in a series of deep eutectic solvents (DESs) composed of acetamide and lithium salts. Quasielastic neutron scattering experiments reveal SEB in the jump diffusion of acetamide, represented by a fractional Stokes-Einstein relationship. Among these DESs, lithium perchlorate exhibits the most pronounced SEB while lithium bromide displays the weakest. Concurrently, elastic incoherent neutron scans identify that bromide DES is the most heterogeneous and perchlorate is the least. For the first time, our study unveils a counterintuitive incommensurate relationship between DH and SEB. Further, it reveals the intricate contrasting nature of the SEB-DH relationship when investigated in proximity to the glass-transition temperature and further away from it.

Diffusion of confined fluids in microporous zeolites and clay materials.

Fluids exhibit remarkable variation in their structural and dynamic properties when they are confined at the nanoscopic scale. Various factors, including geometric restriction, the size and shape of the guest molecules, the topology of the host, and guest-host interactions, are responsible for the alterations in these properties. Due to their porous structures, aluminosilicates provide a suitable host system for studying the diffusion of sorbates in confinement. Zeolites and clays are two classes of the aluminosilicate family, comprising very ordered porous or layered structures. Zeolitic materials are important due to their high catalytic activity and molecular sieving properties. Guest molecules adsorbed by zeolites display many interesting features including unidimensional diffusion, non-isotropic rotation, preferred orientation and levitation effects, depending on the guest and host characteristics. These are useful for the separation of hydrocarbons which commonly exist as mixtures in nature. Similarly, clay materials have found application in catalysis, desalination, enhanced oil recovery, and isolation barriers used in radioactive waste disposal. It has been shown that the bonding interactions, level of hydration, interlayer spacing, and number of charge-balancing cations are the important factors that determine the nature of diffusion of water molecules in clays. Here, we present a review of the current status of the diffusion mechanisms of various adsorbed species in different microporous zeolites and clays, as investigated using quasielastic neutron scattering and classical molecular dynamics simulation techniques. It is impossible to write an exhaustive review of the subject matter, as it has been explored over several decades and involves many research topics. However, an effort is made to cover the relevant issues specific to the dynamics of different molecules in microporous zeolites and clay materials and to highlight a variety of interesting features that are important for both practical applications and fundamental aspects.

Enhanced Microscopic Dynamics of a Liver Lipid Membrane in the Presence of an Ionic Liquid.

Ionic liquids (ILs) are an important class of emerging compounds, owing to their widespread industrial applications in high-performance lubricants for food and cellulose processing, despite their toxicity to living organisms. It is believed that this toxicity is related to their actions on the cellular membrane. Hence, it is vital to understand the interaction of ILs with cell membranes. Here, we report on the effects of an imidazolium-based IL, 1-decyl-3-methylimidazolium tetrafluoroborate (DMIM[BF4]), on the microscopic dynamics of a membrane formed by liver extract lipid, using quasielastic neutron scattering (QENS). The presence of significant quasielastic broadening indicates that stochastic molecular motions of the lipids are active in the system. Two distinct molecular motions, (i) lateral motion of the lipid within the membrane leaflet and (ii) localized internal motions of the lipid, are found to contribute to the QENS broadening. While the lateral motion could be described assuming continuous diffusion, the internal motion is explained on the basis of localized translational diffusion. Incorporation of the IL into the liver lipid membrane is found to enhance the membrane dynamics by accelerating both lateral and internal motions of the lipids. This indicates that the IL induces disorder in the membrane and enhances the fluidity of lipids. This could be explained on the basis of its location in the lipid membrane. Results are compared with various other additives and we provide an indication of a possible correlation between the effects of guest molecules on the dynamics of the membrane and its location within the membrane.

Contrasting Behaviors of FA and MA Cations in APbBr3.

It is known that the organic units in hybrid halide perovskites are free to rotate, but it is not clear if this freedom is of any relevance to the structure-property relationship of these compounds. We have employed quasi-elastic neutron scattering using two different spectrometers, thus providing a wide dynamic range to investigate the cation dynamics in methylammonium lead bromide (MAPbBr3) and formamidinium lead bromide (FAPbBr3) over a large temperature range covering all known crystallographic phases of these two compounds. Our results establish a plastic crystal-like phase forming above 30 K within the orthorhombic phase of MAPbBr3 related to 3-fold rotations of MA units around the C-N axis with an activation energy, Ea, of ∼27 meV, which has no counterpart in the FA compound. MA exhibits an additional 4-fold orientational motion of the whole molecule via rotation of the C-N axis itself with an Ea of ∼68 meV common for the high-temperature tetragonal and cubic phases. In contrast, the FA compound exhibits only an isotropic orientational motion of the whole FA unit with Ea ≈ 106 meV within the orthorhombic phase and a substantially reduced common Ea of ∼62 meV for the high-temperature tetragonal and cubic phases. Our results suggest that the rotational dynamics of the organic units, crystallographic phases, and physical properties of these compounds are intimately connected.

Solvation and transport of lithium ions in deep eutectic solvents.

Lithium based deep eutectic solvents (DESs) are excellent candidates as eco-friendly electrolytes for lithium ion batteries. While some of these DESs have shown promising results, a clear mechanism of lithium ion transport in DESs is not yet established. This work reports the study on the solvation and transport of lithium in a DES made from lithium perchlorate and acetamide using Molecular Dynamics (MD) simulation and inelastic neutron scattering. Based on hydrogen bonding (H-bonding) of acetamide with neighboring molecules/ions, two states are largely prevalent: (1) acetamide molecules that are H-bonded to lithium ions (∼36%) and (2) acetamide molecules that are entirely free (∼58%). Analyzing their stochastic dynamics independently, it is observed that the long-range diffusion of the former is significantly slower than that of the latter. This is also validated from the neutron scattering experiment on the same DES system. Furthermore, the analysis of the lithium dynamics shows that the diffusion of acetamide molecules in the first category is strongly coupled to that of lithium ions. On an average, the lithium ions are H-bonded to ∼3.2 acetamide molecules in their first solvation. These observations are further bolstered through the analysis of the H-bond correlation function between acetamide and lithium ions, which shows that ∼90% of lithium ionic transport is achieved by vehicular motion where the ions diffuse along with their first solvation shell. It is also observed that the ionic motions are largely uncorrelated and the conductivity of lithium ions in the DES is found to be 11 mS/cm. The findings of this work are an important advancement in understanding solvation and transport of lithium in the DES.

Transport Mechanism of Acetamide in Deep Eutectic Solvents.

Over the last couple of decades, deep eutectic solvents (DESs) have emerged as novel alternatives to ionic liquids that are extensively used in the synthesis of innovative materials, metal processing, catalysis, etc. However, their usage is limited, primarily because of their large viscosity and poor conductivity. Therefore, an understanding of the molecular origin of these transport properties is essential to improve their industrial applicability. Here, we present the report of the nanoscopic diffusion mechanism of acetamide in a DES synthesized with lithium perchlorate as studied using neutron scattering and molecular dynamics (MD) simulation techniques. A diffusion model is constructed with the help of MD simulation data comprising two distinct processes, corresponding to long-range jump diffusion and localized diffusion within a restricted volume. This diffusion model is validated through the analysis of neutron scattering data in the acetamide based DES (ADES) and molten acetamide. Although ADES has a remarkably lower freezing point compared to pure acetamide, the molecular mobility is found to be enormously restricted in the former. Particularly, the long-range jump diffusion process of acetamide is slower by a factor of 3 in ADES in comparison with molten acetamide. Further, the geometry of localized diffusion is found to be unaltered, but the dynamics is observed to be slightly slower in ADES. The diffusion model is found to be consistent over a wide temperature range for the ADES. Both long-range and localized diffusion show Arrhenius dependence with temperature in ADES. MD simulation analysis reveals that the long-range diffusion in ADES is restricted mainly due to the formation of hydrogen bond mediated complexes between the ionic species of the salt and acetamide molecules. Hence, the origin of higher viscosity observed in ADES can be attributed to the complexation in the ADES. The complex formation also explains the inhibition of the crystallization process while cooling and thereby results in depression of the freezing point of ADES.

Probing relaxation dynamics of a cationic lipid based non-viral carrier: a time-resolved fluorescence study.

Lipid vesicles composed of cationic dioctadecyldimethylammonium bromide (DODAB) and neutral 1-monooleoyl-rac-glycerol (MO) are promising non-viral carriers of nucleic acids for delivery into cells. Among them, higher cell transfection efficiency was displayed by DODAB-rich vesicles than those enriched with MO. Structural relaxation of these mixed lipid vesicles plays a key role in their cell transfection efficiency because structural organization of the DODAB-rich vesicles are different from that of the MO-rich vesicles. Polarization-gated anisotropy in conjunction with picosecond resolved emission transients of a novel fluorophore 6-acetyl-(2-((4-hydroxycyclohexyl)(methyl)amino)naphthalene) (ACYMAN) has been employed to probe relaxation dynamics in pure DODAB vesicles, and in mixed vesicles of DODAB with varying content of MO. Both orientational relaxation of ACYMAN and relaxation dynamics of its local environment are retarded significantly in mixed lipid vesicles with increasing MO content, from a mole fraction (χ MO) of 0.2 to that of 0.8 which is consistent with increased rigidity of the MO-rich (χ MO > 0.5) vesicles relative to the DODAB-rich (χ MO < 0.5) vesicles. Therefore, higher structural rigidity of the MO-rich vesicles (χ MO > 0.5) gives rise to their lower cell transfection efficiency than the more flexible DODAB-rich (χ MO < 0.5) vesicles as observed in previous in vivo studies (Biochim. Biophys. Acta, Biomembr., 2014, 1838, 2555-2567).

Correction of Toe-Walking Gait in Children with Spastic Cerebral Palsy by using Electrical Stimulation Therapy.

Toe-walking is a very common gait abnormality seen in children with Cerebral Palsy (CP). The present study aims to improvise the toe-walking gait by applying Electrical Stimulation (ES) therapy of the Tricep Surae (TS) muscles. The study was carried out on sixteen children with spastic CP with unilateral toe-walking gait problem, divided into the intervention group that received both ES therapy along with conventional physiotherapy treatment and the control group that received only conventional physiotherapy treatment. Both groups were treated for 60 (30 + 30) minutes per day, for 5 days a week, up to 12 weeks. The gait data were analyzed for spatiotemporal and parameters influencing the walking capacity. The results showed that those children who received the intervention had a significant increase in gait speed by 17.67 % (p = 0.019) and decrease in stride length by 10.25 % (p = 0.037), resulting in improvement of body balance. There was a significant percentage increase in initial contact (heel strike) of 85.71 % (p = 0.000) and flat foot position (loading response) of 49.2 % (p = 0.005), confirming reduction in toe-walking. There was also an increase in the swing power by 39.8 % (p = 0.028) and ground impact by 19.5 % (p = 0.003) suggesting a change in foot contact pattern. The results indicate that ES therapy on TS muscle along with conventional physiotherapy may correct the toe-walking gait in children with spastic hemiplegic CP.

Effects of NSAIDs on the Dynamics and Phase Behavior of DODAB Bilayers.

Despite well-known side effects, nonsteroidal anti-inflammatory drugs (NSAIDs) are one of the most prescribed drugs worldwide for their anti-inflammatory and antipyretic properties. Here, we report the effects of two NSAIDs, aspirin and indomethacin, on the thermotropic phase behavior and the dynamics of a dioctadecyldimethylammonium bromide (DODAB) lipid bilayer as studied using neutron scattering techniques. Elastic fixed window scans showed that the addition of aspirin and indomethacin affects the phase behavior of a DODAB bilayer in both heating and cooling cycles. Upon heating, there is a change in the coagel- to fluid-phase transition temperature from 327 K for pure DODAB bilayer to 321 and 323 K in the presence of aspirin and indomethacin, respectively. More strikingly, upon cooling, the addition of NSAIDs suppresses the formation of the intermediate gel phase observed in pure DODAB. The suppression of the gel phase on addition of the NSAIDs evidences the synchronous ordering of a lipid headgroup and chain. Analysis of quasi-elastic neutron scattering data showed that only localized internal motion exists in the coagel phase, whereas both internal and lateral motions exist in the fluid phase. The internal motion is described by a fractional uniaxial rotational diffusion model in the coagel phase and by a localized translation diffusion model in the fluid phase. In the coagel phase, the rotational diffusion coefficient of DODAB is found to be almost twice for the addition of the drugs, whereas the mobility fraction did not change for indomethacin but becomes twice for aspirin. In the fluid phase, the lateral motion, described well by a continuous diffusion model, is found to be slower by about ∼30% for indomethacin but almost no change for aspirin. For the internal motion, addition of aspirin leads to enhancement of the internal motion, whereas indomethacin did not show significant effect. This study shows that the effect of different NSAIDs on the dynamics of the lipid membrane is not the same; hence, one must consider these NSAIDs individually while studying their action mechanism on the cell membrane.

Modulation of Solvation and Molecular Recognition of a Lipid Bilayer under Dynamical Phase Transition.

It is well accepted in contemporary biology that an ∼30 Šthick lipid bilayer film around living cells is a matter of life and death as the film typically delimits the environments that serve as a crucial margin. The dynamic organization of lipid molecules both across the lipid bilayer and in the lateral dimension are known to be crucial for cellular transport and molecular recognition by important biological macromolecules. Here, we study dilute (20 mM) Dioctadecyldimethylammonium bromide (DODAB) vesicles at different temperatures in aqueous dispersion with well-defined phases namely liquid crystalline, gel and subgel. The spectroscopic studies on two fluorescent probes 8-anilino-1-naphthalene sulfonic acid ammonium salt (ANS) and Coumarin 500 (C500), former in the head group region of the lipid-water interface and later located deeper in the lipid bilayer follow dynamics (solvation and fluidity) of their local environments in the vesicles. Binding of an anti-tuberculosis drug rifampicin has also been studied employing Förster resonance energy transfer (FRET) technique. The molecular insight concerning the effect of dynamical organization of the lipid molecules on the local dynamics of aqueous environments in different phases leading to molecular recognition becomes evident in our study.

Deciphering interactions of ionic liquids with biomembrane.

Ionic liquids (ILs) are a special class of low-temperature (typically < 100 °C) molten salts, which have huge upsurge interest in the field of chemical synthesis, catalysis, electrochemistry, pharmacology, and biotechnology, mainly due to their highly tunable nature and exceptional properties. However, practical uses of ILs are restricted mainly due to their adverse actions on organisms. Understanding interactions of ILs with biomembrane is prerequisite to assimilate the actions of these ionic compounds on the organism. Here, we review different biophysical methods to characterize interactions between ILs and phospholipid membrane, a model biomembrane. All these studies indicate that ILs interact profoundly with the lipid bilayer and modulate the structure, microscopic dynamics, and phase behavior of the membrane, which could be the fundamental cause of the observed toxicity of ILs. Effects of ILs on the membrane are found to be strongly dependent on the lipophilicity of the IL and are found to increase with the alkyl chain length of IL. This can be correlated with the observed higher toxicity of IL with the longer alkyl chain length. These informations would be useful to tune the toxicity of IL which is required in designing environment-friendly nontoxic solvents of the so-called green chemistry for various practical applications.

Negative Differential Resistance Behavior of the Iron Storage Protein Ferritin.

Realization of useful nanometer length scale devices in which metalloproteins are junction-confined in a distinct molecular arrangement for generating practical electronic signals (e.g., in bioelectronic switch configuration) is elusive till date. This is mostly due to difficulties in observing an electronically appropriate signal (i.e., reproducible and controllable), when studied under junction-assembled condition. A useful "ON"-"OFF" behavior, based on the negative differential resistance (NDR) peak characteristics in the current-voltage response curves, acquired using metal-insulator-metal (MIM) configuration, has been observed only in the case of a few proteins, namely, azurin, cytochrome c, bacteriorhodopsin, so far. The case of NDR in ferritin, an iron storage protein having a semiconducting iron core consisting of few thousands of iron atoms connected in an oxide network, has not been studied in the MIM configuration where single (or a few) molecule(s) are junction-trapped, for example, as in the case of local probe configuration of scanning probe microscopy. The present study by scanning tunneling microscopy (STM), using the naturally occurring iron-containing ferritin (human liver), as well as different iron-loaded ferritins, provides clear indication of the capability of ferritins to be NDR capable, at varying sweep conditions. As ferritin can be tailor-made in a structurally conserved manner, metal core-reconstituted ferritins, that is, Mn(III)-ferritin, Cu(II)-ferritin, and Ag-ferritin, were prepared. A correlation between the NDR peak signatures, as observed in the respective current-voltage response curves of these reconstituted ferritins, and the nature of the metal core is demonstrated. In support of our earlier proposition, here, we affirm that the ferritin protein behaves as a conductor-insulator (metal core-polypeptide shell) composite, where the overall electronic structure of the material can alter as a function of the nature of the conducting filler placed inside the insulated matrix.

Dynamical Transitions and Diffusion Mechanism in DODAB Bilayer.

Dioctadecyldimethylammonium bromide (DODAB), a potential candidate for applications in drug transport or DNA transfection, forms bilayer in aqueous media exhibiting a rich phase behavior. Here, we report the detailed dynamical features of DODAB bilayer in their different phases (coagel, gel and fluid) as studied by neutron scattering techniques. Elastic intensity scans show dynamical transitions at 327 K in the heating and at 311 K and 299 K during cooling cycle. These results are consistent with calorimetric studies, identified as coagel-fluid phase transition during heating, and fluid-gel and gel-coagel phase transitions during cooling. Quasielastic Neutron Scattering (QENS) data analysis showed presence of only localized internal motion in the coagel phase. However, in the gel and fluid phases, two distinct motions appear, namely lateral motion of the DODAB monomers and a faster localized internal motion of the monomers. The lateral motion of the DODAB molecule is described by a continuous diffusion model and is found to be about an order of magnitude slower in the gel phase than in the fluid phase. To gain molecular insights, molecular dynamics simulations of DODAB bilayer have also been carried out and the results are found to be in agreement with the experiment.

Effects of ionic liquids on the nanoscopic dynamics and phase behaviour of a phosphatidylcholine membrane.

Ionic liquids (ILs) are potential candidates for new antimicrobials due to their tunable antibacterial and antifungal properties that are required to keep pace with the growing challenge of bacterial resistance. To a great extent their antimicrobial actions are related to the interactions of ILs with cell membranes. Here, we report the effects of ILs on the nanoscopic dynamics and phase behaviour of a dimyristoylphosphatidylcholine (DMPC) membrane, a model cell membrane, as studied using neutron scattering techniques. Two prototypical imidazolium-based ILs 1-butyl-3-methylimidazolium tetrafluoroborate (BMIM[BF4]) and 1-decyl-3-methylimidazolium tetrafluoroborate (DMIM[BF4]), which differ only in terms of the alkyl chain length of cations, have been used for the present study. Fixed Elastic Window Scan (FEWS) shows that the incorporation of ILs affects the phase behaviour of the phospholipid membrane significantly and the transition from a solid gel to a fluid phase shifts to lower temperature. This is found to be consistent with our differential scanning calorimetry measurements. DMIM[BF4], which has a longer alkyl chain cation, affects the phase behaviour more strongly in comparison to BMIM[BF4]. The pressure-area isotherms of the DMPC monolayer measured at the air-water interface show that in the presence of ILs, isotherms shift towards higher area-per lipid molecule. DMIM[BF4] is found to shift the isotherm to a greater extent compared to BMIM[BF4]. Quasielastic neutron scattering (QENS) data show that both ILs act as a plasticizer, which enhances the fluidity of the membrane. DMIM[BF4] is found to be a stronger plasticizing agent in comparison to BMIM[BF4] that has a cation with a shorter alkyl chain. The incorporation of DMIM[BF4] enhances not only the long range lateral motion but also the localised internal motion of the lipids. On the other hand, BMIM[BF4] acts weakly in comparison to DMIM[BF4] and mainly alters the localised internal motion of the lipids. Any subtle change in the dynamical properties of the membrane can profoundly affect the stability of the cell. Hence, the dominant effect of the IL with the longer chain length on the dynamics of the phospholipid membrane might be correlated with its cytotoxic activity. QENS data analysis has provided a quantitative description of the effects of the two imidazolium-based ILs on the dynamical and phase behaviour of the model cell membrane, which is essential for a detailed understanding of their action mechanism.

Effects of Hydrotropic Salt on the Nanoscopic Dynamics of DTAB Micelles.

Effects of a hydrotropic salt, sodium salicylate (NaSal), on the dynamic behavior of cationic dodecyltrimethylammonium bromide (DTAB) micelles as studied using dynamic light scattering (DLS) and quasielastic neutron scattering (QENS) techniques are reported here. DLS study showed that the addition of NaSal leads to a decrease in the apparent diffusion coefficient of the whole micelle indicating micellar growth. QENS data analysis suggested that observed dynamics involves two distinct motions, lateral motion of the surfactant over the curved micellar surface and localized segmental motion of the surfactant. It is found that the addition of NaSal slows down the lateral motion of DTAB while the localized segmental motion of the DTAB chain is not affected much. An atomistic molecular dynamics (MD) simulation was performed to gain further insight into the underlying phenomena. MD simulation results are found to be consistent with the experimental observations. MD simulation revealed that location of the salicylate ions on the micellar surface and their strong electrostatic association with their oppositely charged surfactant headgroup are the major factors in slowing down the lateral motion of the DTAB molecule. In the present work, a quantitative description of the effects of NaSal on the nanoscopic dynamics of DTAB micelles and its correlation with the microstructure of the micelle is provided.

Discriminating Intercalative Effects of Threading Intercalator Nogalamycin, from Classical Intercalator Daunomycin, Using Single Molecule Atomic Force Spectroscopy.

DNA threading intercalators are a unique class of intercalating agents, albeit little biophysical information is available on their intercalative actions. Herein, the intercalative effects of nogalamycin, which is a naturally-occurring DNA threading intercalator, have been investigated by high-resolution atomic force microscopy (AFM) and spectroscopy (AFS). The results have been compared with those of the well-known chemotherapeutic drug daunomycin, which is a non-threading classical intercalator bearing structural similarity to nogalamycin. A comparative AFM assessment revealed a greater increase in DNA contour length over the entire incubation period of 48 h for nogalamycin treatment, whereas the contour length increase manifested faster in case of daunomycin. The elastic response of single DNA molecules to an externally applied force was investigated by the single molecule AFS approach. Characteristic mechanical fingerprints in the overstretching behaviour clearly distinguished the nogalamycin/daunomycin-treated dsDNA from untreated dsDNA-the former appearing less elastic than the latter, and the nogalamycin-treated DNA distinguished from the daunomycin-treated DNA-the classically intercalated dsDNA appearing the least elastic. A single molecule AFS-based discrimination of threading intercalation from the classical type is being reported for the first time.

Enhancement of Lateral Diffusion in Catanionic Vesicles during Multilamellar-to-Unilamellar Transition.

Catanionic vesicles are formed spontaneously by mixing cationic and anionic dispersions in aqueous solution in suitable conditions. Because of spontaneity in formation, long-term stability, and easy modulation of size and charge, they have numerous advantages over conventional lipid-based vesicles. The dynamics of such vesicles is of interest in the field of biomedicine, as they can be used to deliver drug molecules into the cell membrane. Dynamics of catanionic vesicles based on sodium dodecyl sulfate (SDS) and cetyltrimethylammonium bromide (CTAB) have been studied using incoherent elastic and quasielastic neutron scattering (QENS) techniques. Neutron scattering experiments have been carried out on two backscattering spectrometers, IRIS and IN16B, which have different energy resolutions and energy transfer windows. An elastic fixed-window scan carried out using IN16B shows a phase transition at ∼307 K during the heating cycle, whereas on cooling the transition occurred at ∼294 K. DSC results are found to be in close agreement with the elastic scan data. This transition is ascribed to a structural rearrangement from a multilamellar to a unilamellar phase [ Andreozzi J. Phys. Chem. B 2010 , 114 , 8056 - 8060 ]. It is found that a model in which the surfactant molecules undergo both lateral and internal motions can describe the QENS data quite well. While the data from IRIS have contributions from both dynamical processes, the data from IN16B probe only lateral motions, as the internal motions are too fast for the energy window of the spectrometer. It is found that, through the transition, the fraction of surfactant molecules undergoing lateral motion increases of a factor of 2 from the multilamellar to the unilamellar phase, indicating an enhanced fluidity of the latter. The lateral motion is found to be Fickian in nature, while the internal motion has been described by a localized translational diffusion model. The results reported here could have direct interest for a number of applications, such as molecular transport, and the effect of specific drug molecules or hormones through the membrane.

Interactions of Histone Acetyltransferase p300 with the Nuclear Proteins Histone and HMGB1, As Revealed by Single Molecule Atomic Force Spectroscopy.

One of the important properties of the transcriptional coactivator p300 is histone acetyltransferase (HAT) activity that enables p300 to influence chromatin action via histone modulation. p300 can exert its HAT action upon the other nuclear proteins too--one notable example being the transcription-factor-like protein HMGB1, which functions also as a cytokine, and whose accumulation in the cytoplasm, as a response to tissue damage, is triggered by its acetylation. Hitherto, no information on the structure and stability of the complexes between full-length p300 (p300FL) (300 kDa) and the histone/HMGB1 proteins are available, probably due to the presence of unstructured regions within p300FL that makes it difficult to be crystallized. Herein, we have adopted the high-resolution atomic force microscopy (AFM) approach, which allows molecularly resolved three-dimensional contour mapping of a protein molecule of any size and structure. From the off-rate and activation barrier values, obtained using single molecule dynamic force spectroscopy, the biochemical proposition of preferential binding of p300FL to histone H3, compared to the octameric histone, can be validated. Importantly, from the energy landscape of the dissociation events, a model for the p300-histone and the p300-HMGB1 dynamic complexes that HAT forms, can be proposed. The lower unbinding forces of the complexes observed in acetylating conditions, compared to those observed in non-acetylating conditions, indicate that upon acetylation, p300 tends to weakly associate, probably as an outcome of charge alterations on the histone/HMGB1 surface and/or acetylation-induced conformational changes. To our knowledge, for the first time, a single molecule level treatment of the interactions of HAT, where the full-length protein is considered, is being reported.

Direct Observation of Coupling between Structural Fluctuation and Ultrafast Hydration Dynamics of Fluorescent Probes in Anionic Micelles.

The coupling of structural fluctuation and the dynamics of associated water molecules of biological macromolecules is vital for various biological activities. Although a number of molecular dynamics (MD) studies on proteins/DNA predicted the importance of such coupling, experimental evidence of variation of hydration dynamics with controlled structural fluctuation even in model macromolecule is sparse and raised controversies in the contemporary literature. Here, we have investigated dynamics of hydration at the surfaces of two similar anionic micelles sodium dodecyl sulfate (SDS) and sodium dodecylbenzenesulfonate (SDBS) as model macromolecules using coumarin 500 (C500) as spectroscopic probe with femtosecond to picosecond time resolution up to 20 ns time window. The constituting surfactants SDS and SDBS are structurally similar except one benzene moiety in the SDBS may offer additional rigidity to the SDBS micelles through π-stacking and added bulkiness. The structural integrity of the micelles in the aqueous medium is confirmed in dynamic light scattering (DLS) studies. A variety of studies including polarization gated fluorescence spectroscopy and quasielastic neutron scattering (QENS) have been used to confirm differential structural fluctuation of SDS and SDBS micelles. We have also employed femtosecond-resolved Förster resonance energy transfer (FRET) in order to study binding of a cationic organic ligand ethidium bromide (EtBr) salt at the micellar surfaces. The distance distribution of the donor (C500)-acceptor (EtBr) in the micellar media reveals the manifestation of the structural flexibility of the micelles. Our studies on dynamical coupling of the structural flexibility with surface hydration in the nanoscopic micellar media may find the relevance in the "master-slave" type water dynamics in biologically relevant macromolecules.

Structure and dynamics of ionic micelles: MD simulation and neutron scattering study.

Fully atomistic molecular dynamics (MD) simulations have been carried out on sodium dodecyl sulfate (SDS), an anionic micelle, and three cationic (CnTAB; n = 12, 14, 16) micelles, investigating the effects of size, the form of the headgroup, and chain length. They have been used to analyze neutron scattering data. MD simulations confirm the dynamical model of global motion of the whole micelle, segmental motion (headgroup and alkyl chain), and fast torsional motion associated with the surfactants that is used to analyze the experimental data. It is found that the solvent surrounding the headgroups results in their significant mobility, which exceeds that of the tails on the nanosecond time scale. The middle of the chain is found to be least mobile, consolidating the micellar configuration. This dynamical feature is similar for all the ionic micelles investigated and therefore independent of headgroup form and charge and chain length. Diffusion constants for global and segmental motion of the different micelles are consistent with experimentally obtained values as well as known structural features. This work provides a more realistic model of micelle dynamics and offers new insight into the strongly fluctuating surface of micelles which is important in understanding micelle dispersion and related functionality, like drug delivery.