Solution Conformation Research Articles

Osmolyte-induced conformational stabilization of a hydrophobic polymer.

Elucidating the mechanistic role of osmolytes on conformations of hydrophobic prototypical macromolecules in principle is the stepping stone towards understanding the effect of osmolytes on proteins. Motivated by this, we use equilibrium simulations and umbrella sampling techniques to dissect the underlying mechanism of osmolyte-induced conformational stability of a hydrophobic polymer. Our results unveil a remarkable osmolyte-dependent conformational stabilization of the polymer. In an aqueous solution of 4 M choline chloride (ChCl), the polymer has an even more compact structure than in water. On the other hand, an aqueous solution of 8 M urea stabilizes the extended state of the polymer. Interestingly, the polymer adopts an intermediate hairpin conformation in a mixed osmolyte solution of 4 M ChCl and 8 M urea in water due to the interplay of ChCl and urea. Our simulations identify the relative accumulation of water and the hydrophilic part of choline or preferential binding of urea near the collapsed and the extended states, respectively. Analyses split out the enthalpic and entropic contributions to the overall free energy. This decides the stabilization of the preferred conformation in the chosen osmolyte solution. Our simulations show that in an aqueous solution of ChCl, the hairpin state is stabilized by entropy gain. In contrast, the enthalpic contribution stabilizes the hairpin state in mixed environments. However, a collapsed state is energetically not favored in the presence of urea. In brief, via employing an in silico approach, the current findings indicate the importance of osmolytes in stabilizing the conformational states of hydrophobic polymers.

Research progress on the conformational properties of comb-like polymers in dilute solutions.

As a special type of branched polymers, comb-like polymers simultaneously possess the structural characteristics of a linear backbone profile and crowded sidechain branches/grafts, and such structural uniqueness leads to reduced interchain entanglement, enhanced molecular orientation, and unique stimulus-response behavior, which greatly expands the potential applications in the fields of super-soft elastomers, molecular sensors, lubricants, photonic crystals, etc. In principle, all these molecular features can be traced back to three structural parameters, i.e., the degree of polymerization of the backbone (Nb), the degree of polymerization of the graft sidechain (Ng), and the grafting density (σ). Consequently, it is of great importance to understand the correlation mechanism between the structural characteristics and physicochemical properties, among which, the conformational properties in dilute solution have received the most attention due to its central position in polymer science. In the past decades, the development of synthetic chemistry and characterization techniques has greatly stimulated the progress of this field, and a number of experiments have been executed to verify the conformational properties; however, due to the complexity of the structural parameters and the diversity of the chemical design, the achieved experimental progress displays significant controversies compared with the theoretical predictions. This review aims to provide a full picture of recent research progress on this topic, specifically, (1) first, a few classical theoretical models regarding the chain conformation are introduced, and the quasi-two-parameter (QTP) theory for the conformation analysis is highlighted; (2) second, the research progress of the static conformation of comb-like polymers in dilute solution is discussed; (3) third, the research progress of the dynamic conformation in dilute solution is further discussed. The key issues, existing controversies and future research directions are also highlighted. We hope that this review can provide insightful information for the understanding of the conformational properties of comb-like polymers, open a new door for the regulation of conformational behavior in related applications, and promote related theoretical and experimental research in the community.

Conformational control of antimicrobial peptide amphiphilicity: consequences for boosting membrane interactions and antimicrobial effects of photocatalytic TiO2 nanoparticles.

This study reports on the effects of conformationally controlled amphiphilicity of antimicrobial peptides (AMPs) on their ability to coat TiO2 nanoparticles (NPs) and boost the photocatalytic antimicrobial effects of such NPs. For this, TiO2 NPs were combined with AMP EFK17 (EFKRIVQRIKDFLRNLV), displaying a disordered conformation in aqueous solution but helix formation on interaction with bacterial membranes. The membrane-bound helix is amphiphilic, with all polar and charged amino acid residues located at one side and all non-polar and hydrophobic residues on the other. In contrast, the d-enantiomer variant EFK17-d (E(dF)KR(dI)VQR(dI)KD(dF)LRNLV) is unable to form the amphiphilic helix on bacterial membrane interaction, whereas the W-residues in EFK17-W (EWKRWVQRWKDFLRNLV) boost hydrophobic interactions of the amphiphilic helix. Circular dichroism results showed the effects displayed for the free peptide, to also be present for peptide-coated TiO2 NPs, causing peptide binding to decrease in the order EFK17-W > EFK17 > EFK17-d. Notably, the formation of reactive oxygen species (ROS) by the TiO2 NPs was essentially unaffected by the presence of peptide coating, for all the peptides investigated, and the coatings stabilized over hours of UV exposure. Photocatalytic membrane degradation from TiO2 NPs coated with EFK17-W and EFK17 was promoted for bacteria-like model bilayers containing anionic phosphatidylglycerol but suppressed in mammalian-like bilayers formed by zwitterionic phosphatidylcholine and cholesterol. Structural aspects of these effects were further investigated by neutron reflectometry with clear variations observed between the bacteria- and mammalian-like model bilayers for the three peptides. Mirroring these results in bacteria-like model membranes, combining TiO2 NPs with EFK17-W and EFK17, but not with non-adsorbing EFK17-d, resulted in boosted antimicrobial effects of the resulting cationic composite NPs already in darkness, effects enhanced further on UV illumination.

Why phenomenological theory cannot be used to analyze the dynamics of dark photorefractive solitons

The currently used theory of both bright and dark spatial photorefractive (PR) solitons is a phenomenological theory. In the work we consider the dynamics of dark beams using a microscopic model based on the PR transport equations. It is generally believed that such a theory is more accurate. It has been found that these two approaches can give completely different predictions regarding the formation time of dark beams. The discrepancies can reach up to two orders of magnitude. An approximate analytical solution was presented and a new time constant was introduced taking into account crystals from three classes of PR materials: ferroelectrics, sillenites and semiconductors. Full Text: PDF References A. Katti, R.A. Yadav, Optical Spatial Solitons in Photorefractive Materials (Singapore, Springer, 2021) CrossRef E. DelRe, M. Segev, Ch. 23 in Self-focusing: Past and Present, ed. by R.W. Boyd, S.G. Lukishova, Y.R. Shen (New York, Springer, 2009). CrossRef M. Wichtowski, "On conformity of solutions for one-dimensional photorefractive screening solitons with the Kukhtarev–Vinetskii model", Appl. Phys. B 120, 527 (2015). CrossRef M. Segev, B. Crosignani, P. Di Porto et al., in Photorefractive Spatial Solitons, (Kluwer Academic Publisher, New York, 2002). DirectLink K. Zhan, Ch. Hou, S. Pu, "Temporal behavior of spatial solitons in centrosymmetric photorefractive crystals", Opt. Laser Technol. 43, 1274 (2011). CrossRef J. Maufoy, N. Fressengeas, D.Wolfersberger, G. Kugel, "Simulation of the temporal behavior of soliton propagation in photorefractive media", Phys. Rev. E 59, 6116 (1999). CrossRef A. Katti, M.S. Pande, "Formation characteristics of dark solitons in photorefractive crystals having both linear and quadratic electro-optic effect simultaneously", Opt. Quantum Electron. 55, 1132 (2023). CrossRef

Electrostatic-Dominated Conformational Fluctuations and Transition States of Phase Separation in Charge-Balanced Protein Polymer.

Hydration of the protein/polymer is the most important aspect of stability. It is well-known that salts alter the charged polymer's electrostatic forces, ultimately impacting its conformations in solution. The solvent effects lead to certain conformational fluctuations. Previous studies have shown the screening of electrostatic repulsion within the charge-imbalanced protein following charge inversion owing to counterion condensation and phase separation. This article studies conformation stability and phase separation of charge-balanced gelatin (a protein polymer at the isoelectric point) with the addition of different salts. A phenomenon has been reported where the electrostatic effect of salts results in conformational fluctuations in gelatin due to its insufficient hydrations (termed as starvation), which scales with salt concentration. This article also presents different transition states for charge-balanced proteins prior to phase separation. It is concluded that phase separation of a charge-balanced protein passes through a stable state followed by an unstable transition state, where certain unique interactions with salts occur.

An NMR-based approach for the design of crystallization processes for rigid molecules and effects of molecular rigidity and impurities on crystallization pathways

A methodology based on NMR analysis to enable development of crystallization procedures for relatively rigid molecules is presented. The number of relatively stable conformers in solution and their relative proportions are determined from peak splitting of equivalent protons and their relative areas as measured by NMR. Experimental data indicated that slow conformational interconversion was limiting crystallization at low temperatures. Variable temperature NMR analysis was utilized to determine rotamers’ peak coalescence and identify a suitable crystallization solvent. This NMR-based approach led to the development of a manufacturing crystallization recipe of a relatively rigid molecule that was impossible to crystallize via a scalable process. Furthermore, prediction of relatively stable conformer in gas phase, determined by Molecular Dynamics (MD) calculations were in acceptable agreement with data obtained from NMR analyses. Finally, the effects of molecular rigidity and impurities on crystallization and oiling-out pathways are described and an approach to avoid oiling-out is presented.

Exploring Self-Assembled Cationic Nanostructures of Amphiphilic β-Peptides for Amplifying Drug Delivery Efficiency

In this study, a set of short sequences of amphiphilic peptides composed of synthetic amino acids β-alanine and ornithine were designed. The synthesis of these peptides was carried out in a solution phase and then self-assembled into cationic nanostructures. The formation of cationic nanostructures was confirmed using dynamic light scattering (DLS) and the morphology of the nanostructures was studied using transmission electron microscopy (TEM), which revealed that the nanostructure adopted a spherical shape. To gain insight into the conformation of both solid samples and peptide solutions, the Fourier transform infrared (FTIR) and circular dichroism (CD) spectroscopic analysis were performed. Furthermore, the ability of these nanostructures to encapsulate hydrophobic drugs were also investigated. Three different drugs were tested as model drugs: L-dopa, an anti-Parkinson’s drug; curcumin, anti-cancerous drug and tetrabenazine, anti-Huntington’s drug. The nanostructures exhibited the capacity to encapsulate these drugs in varying ratios.

Variation of CI-2 Conformers upon Addition of Methanol to Water: An IMS-MS-Based Thermodynamic Analysis.

Chymotrypsin inhibitor 2 (CI-2) is a well-studied, textbook example of a cooperative, two-state, native ↔ denatured folding transition. A recent hybrid ion mobility spectrometry (IMS)/mass spectrometry (MS) thermal denaturation study of CI-2 (the well-studied truncated 64-residue model) in water reported evidence that this two-state transition involves numerous (∼41) unique native and non-native (denatured) solution conformations. The characterization of so many, often low-abundance, states is possible because of the very high dynamic range of IMS-MS measurements of ionic species that are produced upon electrospraying CI-2 solutions from a variable temperature electrospray ionization source. A thermodynamic analysis of these states revealed large changes in enthalpy (ΔH) and entropy (ΔS) at different temperatures, and it was suggested that such variation might arise because of temperature-dependent conformational changes of the protein in response to changes in the conformational entropy and the dielectric permeability of water, which drops from a value of ε ∼ 79 at 24 °C to ∼ 60 at 82 °C. Herein, we examine how adding methanol to water influences the distributions of CI-2 conformers and their ensuing stabilities. The dielectric constant of a 60:40 water:methanol (MeOH) drops from ε ∼ 60 at 24 °C to ∼ 51 at 64 °C. Although the same set of conformers observed in water appears to be present in 60:40 water:MeOH, the abundance of each is substantially altered by the presence of methanol. Relative free energy values (ΔG) and thermodynamic values [ΔH and ΔS and heat capacities (ΔCp)] are derived from a Gibbs-Helmholtz analysis. A comparison of these data from water and water:MeOH systems allows rare insight into how variations in solvation and temperature affect many-state protein equilibria. While these studies confirm that variations in solvent dielectric constant with temperature affect the distributions of conformers that are observed, our findings suggest that other solvent differences may also affect abundances.

Analytical Modelling of the Slot Opening Function

The slot opening function, also called relative air gap permeance, is a function which, multiplied by the flux density distribution of a slotless geometry, gives the flux density distribution of a slotted configuration. Here, the magnetic field inside the air gap of a multi-slot surface facing a smooth one was studied, by solving the Laplace equation inside the air gap, in terms of a Fourier series. To obtain the Fourier coefficients, at first, the conformal mapping analytical solution of a single-slot configuration along the smooth surface, was considered. Then, the principle of superposition of the single-slot lost flux density distributions was applied to obtain the multi-slot distribution. The approach is valid in general, and in the case of interference among the flux density distributions of adjacent slots, where their mutual effect cannot be neglected. The field distributions obtained by using the proposed slot opening functions were compared with FEM simulations, showing satisfactory agreement. The numerical accuracy limits were also analysed and discussed.

Physicochemical properties and solution conformation of polysaccharides from Toona sinensis (A. Juss) Roem leaves

In this study, two polysaccharide fractions (TSP-1 and TSP-2) were isolated from Toona sinensis leaves. The physicochemical properties and solution conformations of TSP-1 and TSP-2 were investigated. DSC and TG results showed that TSP-1 and TSP-2 had thermal stability. The intrinsic viscosities of TSP-1 and TSP-2 solutions were 11.42 and 6.13 mL/g, respectively. Rheological results showed that the viscosities of TSP-1 and TSP-2 solutions were affected by polysaccharide concentration, Ca2+ and extreme pH. Furthermore, TSP-1 exhibited a weak gel behavior at the concentrations of 0.5 %–2.0 %, while TSP-2 showed a weak gel behavior at the concentration of 2 %. HPSEC-MALLS analysis revealed that the Rg values of TSP-1 and TSP-2 were 96.8 nm and 56.2 nm, respectively. Conformation analysis indicated that TSP-1 behaved as a sphere, while TSP-2 behaved like a rigid rod. These results suggest that TSP-1 and TSP-2 can be used as additives in food, pharmaceutical and cosmetic industries.

Energy condition of wormhole and thin shell by assuming conformal symmetry in teleparallel-Rastall gravity

The static spherically symmetric wormhole solution for a non-isotropic perfect fluid energy-momentum tensor within the context of teleparallel-Rastall gravity is investigated. First, by imposing static and non-static conformal symmetry on the static spherically symmetric space-time metric, we investigated the null energy condition at the throat of traversable wormholes and showed that the null energy condition in the throat and its neighborhood is violated. Secondly, with the aim of minimize the usage of exotic matter, we found regions where the energy conditions are valid at the junction surface. We found that in the static case of conformal symmetry solution, the null energy condition (NEC), weak energy condition (WEC), and Strong energy condition (SEC) are valid for different λ 0 parameters. Moreover, we obtained that for the non-static symmetric solution of conformal symmetry, the NEC and WEC are satisfied for different sets of parameters. Finally, we have explored the stable regions by the parameter η 2 (square speed of sound) for different sets of parameters.

Analytical Ultracentrifugation Detects Quaternary Rearrangements and Antibody-Induced Conformational Selection of the SARS-CoV-2 Spike Trimer.

Analytical ultracentrifugation (AUC) analysis shows that the SARS-CoV-2 trimeric Spike (S) protein adopts different quaternary conformations in solution. The relative abundance of the "open" and "close" conformations is temperature-dependent, and samples with different storage temperature history have different open/close distributions. Neutralizing antibodies (NAbs) targeting the S receptor binding domain (RBD) do not alter the conformer populations; by contrast, a NAb targeting a cryptic conformational epitope skews the Spike trimer toward an open conformation. The results highlight AUC, which is typically applied for molecular mass determination of biomolecules as a powerful tool for detecting functionally relevant quaternary protein conformations.

Quantum chemical investigation of predominant conformation of the antibiotic azithromycin in water and DMSO solutions: thermodynamic and NMR analysis.

Azithromycin (AZM) is a macrolide-type antibiotic used to prevent and treat serious infections (mycobacteria or MAC) that significantly inhibit bacterial growth. Knowledge of the predominant conformation in solution is of fundamental importance for advancing our understanding of the intermolecular interactions of AZM with biological targets. We report an extensive density functional theory (DFT) study of plausible AZM structures in solution considering implicit and explicit solvent effects. The best match between the experimental and theoretical nuclear magnetic resonance (NMR) profiles was used to assign the preferred conformer in solution, which was supported by the thermodynamic analysis. Among the 15 distinct AZM structures, conformer M14, having a short intramolecular C6-OH … N H-bond, is predicted to be dominant in water and dimethyl sulfoxide (DMSO) solutions. The results indicated that the X-ray structure backbone is mostly conserved in solution, showing that large flexible molecules with several possible conformations may assume a preferential spatial orientation in solution, which is the molecular structure that ultimately interacts with biological targets.

Human Aha1's N-terminal extension confers it holdase activity in vitro.

Molecular chaperones are key components of protein quality control system, which plays an essential role in controlling protein homeostasis. Aha1 has been identified as a co-chaperone of Hsp90 known to strongly accelerate Hsp90's ATPase activity. Meanwhile, it is reported that Aha1 could also act as an autonomous chaperone and protect stressed or disordered proteins from aggregation. Here, in this article, a series of in vitro experiments were conducted to verify whether Aha1 has a non-Hsp90-dependent holdase activity and to elucidate the associated molecular mechanism for substrate recognition. According to the results of the refolding assay, the highly conserved N-terminal extension spanning M1 to R16 in Aha1 from higher eukaryotes is responsible for the holdase activity of the protein. As revealed by the NMR data, Aha1's N-terminal extension mainly adopts a disordered conformation in solution and shows no tight contacts with the core structure of Aha1's N-terminal domain. Based on the intrinsically disordered structure feature and the primary sequence of Aha1's N-terminal extension, the fuzzy-type protein-protein interactions involving this specific region and the unfolded substrate proteins are expected. The following mutation analysis data demonstrated that the Van der Waals contacts potentially involving two tryptophans including W4 and W11 do not play a dominant role in the interaction between Aha1 and unfolded maltose binding protein (MBP). Meanwhile, since the high concentration of NaCl could abolish the holdase activity of Aha1, the electrostatic interactions mediated by those charged residues in Aha1's N-terminal extension are thus indicated to play a crucial role in the substrate recognition.

Structure of Cellulose Ether Affected by Ionic Surfactant and Solvent: A Small-Angle Neutron Scattering Investigation.

Polysaccharides and their derivatives are commonly used in pharmaceutical and agricultural formulations as rheology modifiers. Their performance is related to their conformation in solution, which in turn is affected by other ingredients present in the formulation. This study focuses on modulating the conformation of relatively rigid cellulose chains in aqueous solutions. In particular, we have investigated the nonionic cellulose derivative ethyl(hydroxyethyl)cellulose (EHEC) in water in the presence of the ionic surfactant sodium dodecyl sulfate (SDS) and/or ethanol acting as modulating agents. We have used small angle neutron scattering (SANS) with contrast variation to determine the EHEC chain conformation in the presence of (but not masked by) ethanol and SDS. In dilute and semidilute aqueous solutions, EHEC exhibits worm-like chain conformation due to the rigid cellulose backbone. Addition of ethanol does not impact the polymer conformation to a great extent. Addition of SDS alters the EHEC chain conformation, resulting in polyelectrolyte-like scattering behavior due to repulsive interactions between bound charged micelles which show similar structure as the free SDS micelles in solution (in the absence of polymers). Ethanol affects the polymer + surfactant system primarily by acting on the surfactant (bound on polymer) which, in turn, affects the polymer conformation. At higher ethanol concentrations (20 wt %), EHEC regains the worm-like chain conformation because of the detachment of the bound SDS micelles. To the best of our knowledge, this is the only study providing details on chain conformation of the rigid polymer EHEC in dilute or semidilute aqueous solutions in the presence of surfactant and alcohol and one of very few papers utilizing SANS for the characterization of polymer + surfactant + water + alcohol interactions. Such fundamental understanding of interactions and structure in multicomponent mixtures supports the design of industrial formulations.

MA'AT Analysis: Probability Distributions of Molecular Torsion Angles in Solution from NMR Spectroscopy.

ConspectusMonosaccharides adopt multiple conformations in solution, and this structural complexity increases significantly when they are assembled into oligosaccharides and polysaccharides. Characterization of the conformational properties of saccharides in solution by NMR spectroscopy has been hampered by several complicating factors, including difficulty interpreting spectra because of significant signal overlap, population averaging of NMR parameters, and unique properties of the spectra that make accurate measurements of NMR parameters prone to error (e.g., non-first-order effects on J-couplings). Current conformational assignments rely heavily on theoretical calculations, especially molecular dynamics (MD) simulations, to interpret the experimental NMR parameters. While these studies assert that the available experimental data fit the calculated models well, a lack of independent experimental validation of the force fields from which MD models are derived and an inability to test all possible models that might be compatible with the experimental data in an unbiased manner make the approach less than ideal.NMR spin couplings or J-couplings have been used as structure constraints in organic and other types of molecules for more than six decades. The dihedral angle dependence of vicinal (three-bond) 1H-1H spin couplings (3JHH) first described by Karplus led to an explosion of applications for a wide range of conformational problems. Other vicinal J-couplings (e.g., 3JCCOP, 3JHCOP, and 3JCOCH) have been found to exhibit similar dihedral angle dependencies. 3J values have been used to assign the preferred conformation in molecules that are conformationally homogeneous. However, many molecules, particularly those in biological systems, are conformationally flexible, which complicates structural interpretations of J values in solution. Three-state staggered models are often assumed in order to deconvolute the conformationally averaged J values into conformer populations. While widely applied, this approach assumes highly idealized models of molecular torsion angles that are likely to be poor representations of those found in solution. In addition, this treatment often gives negative populations and neglects the presence of librational averaging of molecular torsion angles.Recent work in this research group has focused on the development of a hybrid experimental-computational method, MA'AT analysis, that provides probability distributions of molecular torsion angles in solution that can be superimposed on those obtained by MD. Ensembles of redundant NMR spin couplings, including 3J (vicinal), 2J (geminal), and sometimes 1J (direct) values, are used in conjunction with circular statistics to provide single- and multistate models of these angles. MA'AT analysis provides accurate mean torsion angles and circular standard deviations (CSDs) of each mean angle that describe the librational motion about the angle. Both conformational equilibria and dynamics are revealed by the method. In this Account, the salient features of MA'AT analysis are discussed, including some applications to conformational problems involving saccharides and peptides.

The antimicrobial peptide chensinin-1b alleviates the inflammatory response by targeting the TLR4/NF-κB signaling pathway and inhibits Pseudomonas aeruginosa infection and LPS-mediated sepsis

Excessive inflammatory responses are a major contributor to the high mortality associated with sepsis, a prevalent global complication. Therefore, the potential therapeutic strategy for sepsis involves targeting macrophages and reducing proinflammatory cytokine release. Chensinin-1b, an analog of the natural antimicrobial peptide derived from Rana chensinensis skin secretion, exhibits broad-spectrum antibacterial activity and adopts a random coil conformation in both PBS and membrane solution. By efficiently neutralizing LPS, chensinin-1b holds promise in alleviating LPS-induced inflammatory responses. In this study, we established a mouse septic shock model by exposing mice to multiple-drug-resistant Pseudomonas aeruginosa, as well as an endotoxin-mediated sepsis model induced by LPS. Administering chensinin-1b significantly prolonged the survival of the experimental mice, concurrently mitigating inflammatory responses and reducing organ damage. Additionally, we investigated the anti-inflammatory mechanism of chensinin-1b using a constructed LPS-induced mouse macrophage RAW264.7 inflammatory model. Our findings demonstrated that chensinin-1b effectively mitigated the excessive activation of the TLR4/NF-κB signaling pathway by directly neutralizing extracellular LPS, thus ameliorating the inflammatory response. Moreover, upon blocking the TLR4 signaling pathway, chensinin-1b further reduced the release of proinflammatory cytokines induced by LPS, indicating alternative modes of regulation. Notably, chensinin-1b rapidly entered RAW264.7 cells within 30 min via endocytosis, diffusing into the cytoplasm while retaining its anti-inflammatory properties intracellularly. Although further investigations are warranted to comprehensively elucidate the intracellular anti-inflammatory mechanism of chensinin-1b, our findings substantiate its possession of anti-inflammatory properties both intracellularly and extracellularly. Thus, chensinin-1b emerges as a promising candidate for mitigating excessive inflammatory responses associated with sepsis.

Dectin-1 multimerization and signaling depends on fungal β-glucan structure and exposure

Dectin-1A is a C-type lectin innate immunoreceptor that recognizes β-(1,3;1,6)-glucan, a structural component of Candida species cell walls. β-Glucans can adopt solution structures ranging from random coil to insoluble fiber due to tertiary (helical) and quaternary structure. Fungal β-glucans of medium and high molecular weight are highly structured, but low molecular weight glucan is much less structured. Despite similar affinity for Dectin-1, the ability of glucans to induce Dectin-1A-mediated signaling correlates with degree of structure. Glucan denaturation experiments showed that glucan structure determines agonistic potential, but not receptor binding affinity. We explored the impact of glucan structure on molecular aggregation of Dectin-1A. Stimulation with glucan signaling decreased Dectin-1A diffusion coefficient. Fluorescence measurements provided direct evidence of ligation-induced Dectin-1A aggregation, which positively correlated with increasing glucan structure content. In contrast, Dectin-1A is predominantly in a low aggregation state in resting cells. Molecular aggregates formed during interaction with highly structured, agonistic glucans did not exceed relatively small (<15 nm) clusters of a few engaged receptors. Finally, we observed increased molecular aggregation of Dectin-1A at fungal particle contact sites in a manner that positively correlated with the degree of exposed glucan on the particle surface. These results indicate that Dectin-1A senses the solution conformation of β-glucans through their varying ability to drive receptor dimer/oligomer formation and activation of membrane proximal signaling events.

Evaluation of nonnatural L-iminosugar C,C-glycosides, a new class of C-branched iminosugars, as glycosidase inhibitors

Capitalizing on a previously developed Staudinger/azaWittig/Grignard (SAWG)-ring contraction sequence that furnished protected six-membered L-iminosugar C,C-glycosides bearing an allyl group and various substituents at the pseudoanomeric position, the synthesis and glycosidase inhibition of a small library of six- and seven-membered L-iminosugar C,C-glycosides is reported. Their hydrogenolysis or cyclization by RCM followed by deprotection afforded eleven L-iminosugars including spirocyclic derivatives. All compounds adopt a 1C4 conformation in solution according to NMR data. Compared to previously reported branched L-iminosugars, the L-iminosugar C,C-glycosides reported herein were less potent glycosidase inhibitors. However, some of these compounds showed micromolar inhibition of human lysosome β-glucocerebrosidase suggesting that such iminosugars could be useful to access potent CGase inhibitors by adjusting the structure/length of the pseudoanomeric substituents.

Conformationally constrained cyclic grafted peptidomimetics targeting protein-protein interactions.

Sunflower trypsin inhibitor-1 (SFTI-1) structure is used for designing grafted peptides as a possible therapeutic agent. The grafted peptide exhibits multiple conformations in solution due to the presence of proline in the structure of the peptide. To lock the grafted peptide into a major conformation in solution, a dibenzofuran moiety (DBF) was incorporated in the peptide backbone structure, replacing the Pro-Pro sequence. NMR studies indicated a major conformation of the grafted peptide in solution. Detailed structural studies suggested that SFTI-DBF adopts a twisted beta-strand structure in solution. The surface plasmon resonance analysis showed that SFTI-DBF binds to CD58 protein. A model for the protein-SFTI-DBF complex was proposed based on docking studies. These studies suggested that SFTI-1 grafted peptide can be used to design stable peptides for therapeutic purposes by grafting organic functional groups and amino acids. However, when a similar strategy was used with another grafted peptide, the resulting peptide did not produce a single major conformation, and its biological activity was lost. Thus, conformational constraints depend on the sequence of amino acids used for SFTI-1 grafting.