Study of the cholesterol-GM3 ganglioside interaction by surface pressure measurements and fluorescence microscopy

The nature of the cholesterol/glycolipid interaction in rafts being poorly understood, the interaction of cholesterol with the GM₃ ganglioside has been studied by surface pressure measurements and fluorescence microscopy. Results have been compared to those obtained with sphingomyelin (SM)-cholesterol and palmitoyl-oleoyl-phosphatidylcholine (POPC)–cholesterol monolayers. The analysis of (π–A) isotherms of mixed monolayers show a condensing effect of cholesterol on GM₃ molecules, in the same range than the effect observed with POPC and higher than the effect on SM. This is likely due to the similar state of GM₃ and POPC, since both molecules are in liquid expanded phases in our experimental conditions. The study of the cholesterol desorption induced by β-cyclodextrin suggests also that the GM₃–cholesterol interaction is rather weak as in the case of POPC–cholesterol interaction, and clearly lower than SM–cholesterol one. This lack of interaction is discussed in terms of nature of lipid chains and molecular shape, and suggests that no hydrogen bond is formed between GM₃ and cholesterol polar heads. Fluorescence microscopy performed on mixed GM₃–cholesterol monolayers shows the presence, at surface pressure higher than 10 mN/m, of particular blurring patterns without defined boundary, which could be due to a partial solubilization in one phase of different phases observed at lower surface pressure, whereas SM–cholesterol and POPC–cholesterol monolayers are homogeneous at the lateral resolution of our microscopy set-up.     

Heat-stabilized glycosphingolipid films for biosensing applications

We have investigated a means of producing thin, oriented lipid monolayers which are stable under repeated washing and which may be useful in biosensing or surface-coating applications. Phosphatidylcholine and the glycosphingolipid GM1 were used as representative lipids for this work. Initially, a mixed self-assembled monolayer of octanethiol and hexadecanethiol was produced on a gold surface. This hydrophobic monolayer was then brought into contact with a thin lipid film that had been assembled at the liquid/air interface of a solution, allowing the lipid to deposit on the gold surface through hydrophobic interactions. The lipid layer was then heated to cause intermingling of the fatty acid and alkanethiol chains and cooled to form a highly stable film which withstood repeated rinsing and solution exposure. Presence and stability of the film were confirmed via ellipsometry, Fourier transform infrared spectroscopy, and quartz crystal microbalance (QCM), with an average overall film thickness of approximately 3.5 nm. This method was then utilized to produce GM1 layers on gold-coated QCM crystals for affinity sensing trials with cholera toxin. For these sensing elements, the lower detection limit of cholera toxin was found to be approximately 0.5 microg/mL, with a logarithmic relationship between toxin concentration and frequency response spanning over several orders of magnitude. Potential sites for nonspecific adsorption were blocked using serum albumin without sacrificing toxin specificity.     

Colloid Stabilization by an Oppositely Charged Polysaccharide: Mechanism of Interaction and Interface Studied with Synchrotron X-Rays

The liposome surface is modeled by a 2-D lipid monolayer made of behenic acid forming a negatively charged interface. The properties at the air/liquid interface were examined by pressure-area isotherms in a Langmuir trough introducing diluted chitosan solution in the subphase. X-ray reflectivity of the interface was measured in the same conditions in order to determine the layer thickness of the chitosan adsorbed on the behenic acid monolayer formed on water. Influence of pH of the subphase and molecular weight of adsorbed chitosan was investigated. All these results allow confirming that the charge and the stability of the lipid layer are controlled by the nature of the polyelectrolyte and the nature of the medium. In particular, the use of biocompatible charged polysaccharides is of interest for biomedical applications.     

Structural characterization of the smectic polymorphism of chiral liquid crystalline polysiloxanes containing biphenyl mesogens

A structural characterization of chiral side chain siloxanes with different average degrees of polymerization, DP_n, was performed by X-ray diffraction experiments on powder and oriented fibre specimens. Polymers (DP_n = 35) and oligomers (DP_n = 4) contained the 4,4'-biphenylene unit with either an (S)-2-methylbutoxy (An, Bn) or an (S)-2-chloro-3-methylbutanoyloxy substituent (Cll). The spacer segment connected to the siloxane backbone had a variable number, n, of methylene groups (n = 5, 8, or 11). Independent of the spacer length and the chiral tail nature, the polysiloxanes underwent the same sequence of phases: C-S_F1 (or S_I1)-S_C1-S_A1-I, whereas in the oligosiloxanes the sequence C-S_B1-S_A1-I (B11) or C-S_F1-S_C1-I (B5) occurred. The influence of the structure of the polysiloxanes on the formation of the smectic (tilted or orthogonal) mesophases was elucidated. The rather large number of reflections (three or four) detected in the X-ray patterns at low angles, allowed a drawing of the projection of the electron density profiles along the layer normal, p(z), and deduction of the most physically acceptable electron density profiles from among the numerous possibilities for each smectic phase. The electron density profiles were in agreement with monolayer smectic phases presenting a microphase separation between the siloxane backbones and the side chains, so constraining the polymer backbones within a thin layer.     

Lipid lateral mobility and membrane phase structure modulation by protein binding

Using a combination of fluorescence correlation and infrared absorption spectroscopies, we characterize lipid lateral diffusion and membrane phase structure as a function of protein binding to the membrane surface. In a supported membrane configuration, cholera toxin binding to the pentasaccharaide headgroup of membrane-incorporated GM1 lipid alters the long-range lateral diffusion of fluorescently labeled probe lipids, which are not involved in the binding interaction. This effect is prominently amplified near the gel-fluid transition temperature, Tm, of the majority lipid component. At temperatures near Tm, large changes in probe lipid diffusion are measured at average protein coverage densities as low as 0.02 area fraction. Spectral shifts of the methylene symmetric and asymmetric stretching modes in the lipid acyl chain confirm that protein binding alters the fraction of lipid in the gel phase.     

Synthesis and characterisation of the methylene-inserted mixed-chalcogenide compounds Fe2(CO)6(μ-SeCH2Te) and Fe2(CO)6(μ-SCH2Te)

The methylene-bridged, mixed-chalogen compounds Fe₂(CO)₆(μ-SeCH₂Te) (1) and Fe₂(CO)₆(μ-SCH₂Te) (3) have been synthesised from the room temperature reaction of diazomethane with Fe₂(CO)₆(μ-SeTe) and Fe₂(CO)₆(μ-STe), respectively. Compounds 1 and 3 have been characterised by IR, ¹H, ¹³C, ⁷⁷Se and ¹²⁵Te NMR spectroscopy. The structure of 1 has been elucidated by X-ray crystallography. The crystalsare monoclinic,space group P2₁/n, A = 6.695(2), B = 13.993(5), C = 14.007(4)Å, β = 103.03(2)°, V = 1278(7) ų, Z = 4, D_c = 2.599 g cm⁻³ and R = 0.030 (R_w = 0.047).     

Structural investigation of dibromobis(benzimidazole)Zn(II) complex

The molecular structure of the title complex [ZnBr₂(C₇H₆N₂)₂] was investigated by X-ray diffraction and IR spectroscopy methods. Molecules of zinc(II) complex crystallize in the triclinic crystal system with cell constants a=7.526(2) Å, b=7.8971(8) Å, c=13.431(1) Å, Z=2 and V=791.3(2) Å³. In the molecular structure, the Zn atom is coordinated tetrahedrally by two Br⁻ anions and two benzimidazole ligands. Intramolecular steric repulsions between Br⁻ anions and benzimidazole groups have been caused to cis configuration around the central metal atom.     

Structure of liquid 1-chloronaphthalene at 293 K

The structure of 1-chloronaphthalene, C₁₀H₇–Cl, at 293 K was investigated using the X-ray diffraction method. Monochromatic radiation MoK (λ=0.71069 Å) enabled the determination of the scattered radiation intensity between S₀=4πsin ₀/λ=0.430 Å⁻¹ and S_max=14.311 Å⁻¹. The interpretation of the results was carried out using the reduction method of Blum and Narten. Experimental distributions of X-ray scattered intensity were compared with theoretical results predicted for a proposed model of 1-chloronaphthalene molecule. The electron-density radial-distribution function was calculated and some intra- and intermolecular distances in liquid 1-chloronaphthalene were determined. X-ray structural analysis was applied to determine the packing coefficient of 1-chloronaphthalene molecules.     

Phase Diagram of RbI-SmI_2 Binary System and Structural Investigation of RbSm_2I_5 and RbSmI_3

The RbI-SmI2 , binary system was studied with DTA and X-ray powder diffraction. Its phase diagram shows that it has a congruently melting compound,RbSm2I5 , with a melting point 524℃; an incongru-ently melting compound, Rb2SmI4 , with an inconsruently point 467℃ and two compounds, RbSmI3, and Rb3SmI5, being decomposed at 433℃ and 460℃, respectively. The measured structure of RbSm2I5 shows that it is isomorphous with NH4 Pb2 Cl5 space group P21/c, a=10 . 063(3)A, b=9.003(3)A. c =14 . 242(5)A°,β=90.23(3)°;z = 4, MVx= 1 94 . 3cm3/mol;D=5.255g/cm3 and Do =5. 334g/cm3 (25. 0℃).The structural, investigation ofRbSmI3 shows that it belongs to the orthorhombic system with a = 15.914(6)A, b=8.898(3)A c =12.193(5)A; Z=8, MVx =130.0cm3/mol, Dx =4 . 743g/cm3,Do = 4.597g/cm3(25.0℃).     

Syntheses, crystal structures and magnetic properties of Ni(II)–2,4-pyridine-dicarboxylates

Two new complexes [Ni(pydc)(H₂O)₂]_n (1) and [Ni₂(pydc)₂(H₂O)₅]_n (2) (H₂pydc = 2,4-pyridinedicarboxylic acid) have been obtained by hydrothermal synthetic method and characterized by single crystal X-ray analysis. In 1 six-coordinate Ni(II) ions are coordinated by pydc ligands to form 2-D layer structures; while in 2 six-coordinate Ni(II) ions are only connected into 1-D zigzag chains constructed by dinuclear nickel units. Although the coordination geometries around Ni(II) centers in both complexes are similar, their structure topologies are greatly tuned by coordination modes of pydc. Variable temperature magnetic susceptibility studies have shown that both compounds 1 and 2 may display antiferromagnetic coupling between paramagnetic metal centers mediated by bridging carboxylate groups.     

Tautomeric properties, conformations and structure of N-(2-hydroxy-5-chlorophenyl) salicylaldimine

The crystal structure of N-(2-hydroxy-5-chlorophenyl) salicylaldimine (C₁₃H₁₀NO₂Cl) was determined by X-ray analysis. It crystallizes orthorhombic space group P2₁2₁2₁ with a=12.967(2) Å, b=14.438(3) Å, c=6.231(3) Å, V=1166.5(6) Å³, Z=4, D_c=1.41 g cm⁻³ and μ(MoK)=0.315 mm⁻¹. The title compound is thermochromic and the molecule is nearly planar. Both tautomeric forms (keto and enol forms in 68(3) and 32(3)%, respectively) are present in the solid state. The molecules contain strong intramolecular hydrogen bonds, N1–H1O1/O2 (2.515(1) and 2.581(2) Å) for the keto form and O1–H01N1 for the enol one. There is also strong intermolecular O2–HO1 hydrogen bonding (2.599(2) Å) between neighbouring molecules. Minimum energy conformations AM1 were calculated as a function of the three torsion angles, θ₁(N1–C7–C6–C5), θ₂(C8–N1–C7–C6) and θ₃(C9–C8–N1–C7), varied every 10°. Although the molecule is nearly planar, the AM1 optimized geometry of the title compound is not planar. The non-planar conformation of the title compound corresponding to the optimized X-ray structure is the most stable conformation in all calculations.     

Tautomeric properties, conformations and structure of N-(2-hydroxyphenyl)-4-amino-3-penten-2-on

N-(2-hydroxyphenyl)-4-amino-3-penten-2-on (C₁₁H₁₃NO₂) has been studied by X-ray analysis. It crystallizes the orthorhombic space group P2₁2₁2₁ with a=8.834(1), b=10.508(2), c=11.212(2) Å, V=1040.8(3) Å³, Z=4, D_c=1.22 g cm⁻³ and μ(MoK)=0.084 mm⁻¹. The structure was solved by direct methods and refined to R=0.038 for 1373 reflections (I>2σ(I)). The title compound is photochromic and the molecule is not planar. Intramolecular hydrogen bonds occur between the pairs of atoms N(1) and O(1) [2.631(2) Å], and N(1) and O(2) [2.641(2) Å], the H atom essentially being bonded to the N atom. There is also a strong intermolecular O–HO hydrogen bonding [2.647(2) Å] between neighbouring molecules. Tautomeric properties and conformations of the title compound were investigated by semi-empirical quantum mechanical AM1 calculations and the results are compared with the X-ray results.     

Modeling the Binding of Cholera Toxin to a Lipid Membrane by a Non‐additive Two‐Dimensional Hard‐Disk Model

A coupled two‐layer hard‐disk model in two dimensions inspired by the interactions of mixtures of different lipids in the outer bilayer leaflet was derived. It is effectively a non‐additive mixture of hard disks. A gas and a crystal phase are observed. The recognition and binding of pathogenic cholera toxin are modeled by fixing lipid‐binding sites in space in pentagonal geometries constrained by the protein. At constant pressure, this positional constraint leads to an increase in the area per molecule as the lipid‐packing structure is distorted. This distortion increases with increasing density or pressure. Variations of the basic model are discussed. We find that the overall area per molecule scales with the area per molecule of the majority component.     

Surface immobilization of bio-functionalized cubosomes: sensing of proteins by quartz crystal microbalance

A strategy for tethering lipid liquid crystalline submicrometer particles (cubosomes) to a gold surface for the detection of proteins is reported. Time-resolved quartz crystal microbalance (QCM-D) was used to monitor the cubosome-protein interaction in real time. To achieve specific binding, cubosomes were prepared from the nonionic surfactant phytantriol, block-copolymer, Pluronic F-127, and a secondary biotinylated lipid, 1,2-distearoyl-sn-glycero-3-phosphoethanolamine-N-[biotinyl(polyethyleneglycol)-2000], which enabled attachment of the particles to a neutravidin (NAv)-alkanethiol monolayer at the gold surface of the QCM sensor chip. A second set of cubosomes was further functionalized with addition of the glycolipid (G(M1)) to facilitate a specific binding uptake of the protein, cholera toxin B subunit (CT(B)), from solution. QCM-D confirmed the specificity of the cubosome-NAv binding. The analysis of titration experiments, also performed with QCM, suggests that an optimal concentration of cubosomes is required for the efficient packing of the particles at the surface: high cubosome concentrations lead to chaotic cubosome binding onto the surface, sterically inhibiting surface attachment, or require significant reorganization to permit uniform cubosome coverage. The methodology enabled the straightforward preparation of a complex nanostructured edifice, which was then used to specifically capture analyte proteins (cholera toxin B subunit or free NAv) from solution, supporting the potential for development of this approach as a biosensing platform.     

Ni在ZrO2(111)薄膜表面的生长、电子结构及热稳定性

利用X射线光电子能谱、紫外光电子能谱和低能电子衍射研究了Ni纳米颗粒在ZrO₂(111)薄膜表面的生长模式、电子结构及热稳定性. ZrO₂(111)薄膜外延生长于Pt(111)单晶表面,厚度约为3 nm.结果表明,当Ni气相沉积到ZrO₂(111)薄膜表面上时,遵循Stranski-Krastanov生长模式,即先二维生长至0.5 ML(monolayer),然后呈三维岛状生长.随着覆盖度的减小, Ni 2p_3/2峰逐渐向高结合能位移.利用俄歇参数法分析发现,引起该峰向高结合能位移的主要原因来源于终态效应的贡献,但在低的Ni覆盖度时,也有部分初态效应的贡献,说明Ni在ZrO₂表面初始生长时,两者存在较强的界面相互作用, Ni向ZrO₂衬底传递电荷,形成带部分正电荷的Ni_δ+.两种不同覆盖度(0.05和0.5 ML)的Ni/ZrO₂(111)模型催化剂热稳定性研究表明,当温度升高时, Ni逐渐被氧化成Ni₂₊,并伴随着向ZrO₂衬底的扩散.本文从原子水平上认识了Ni与ZrO₂表面的相互作用和界面结构,为理解实际ZrO₂担载的Ni催化剂结构提供了重要的依据.     

Solution and crystallographic studies of branched multivalent ligands that inhibit the receptor-binding of cholera toxin

The structure-based design of multivalent ligands offers an attractive strategy toward high affinity protein inhibitors. The spatial arrangement of the receptor-binding sites of cholera toxin, the causative agent of the severe diarrheal disease cholera and a member of the AB(5) bacterial toxin family, provides the opportunity of designing branched multivalent ligands with 5-fold symmetry. Our modular synthesis enabled the construction of a family of complex ligands with five flexible arms each ending with a bivalent ligand. The largest of these ligands has a molecular weight of 10.6 kDa. These ligands are capable of simultaneously binding to two toxin B pentamer molecules with high affinity, thus blocking the receptor-binding process of cholera toxin. A more than million-fold improvement over the monovalent ligand in inhibitory power was achieved with the best branched decavalent ligand. This is better than the improvement observed earlier for the corresponding nonbranched pentavalent ligand. Dynamic light scattering studies demonstrate the formation of concentration-dependent unique 1:1 and 1:2 ligand/toxin complexes in solution with no sign of nonspecific aggregation. This is in complete agreement with a crystal structure of the branched multivalent ligand/toxin B pentamer complex solved at 1.45 A resolution that shows the specific 1:2 ligand/toxin complex formation in the solid state. These results reiterate the power of the structure-based design of multivalent protein ligands as a general strategy for achieving high affinity and potent inhibition.     

The synthesis, X-ray and DFT structures of the free ansa–cyclopentadiene ligand C5H5CMe2CMe2C5H5

The title compound 2,3-dicyclopentadiene-2,3-dimethylbutane (C₅H₅CMe₂CMe₂C₅H₅) 1 shows the typical staggered conformation of a highly substituted ethane derivative with the two largest substituents (C₅H₅) adopting a trans position. The molecule shows C₂ symmetry about the central C–C bond. Due to the high substitution, the central bond of the ethane is elongated to 160.0 pm (X-ray structure analysis) while the DFT calculation finds a value of 159.2 pm.     

Reactions of nickelocene with methyl-, ethyl- and vinyl-lithium compounds. Hydrogen elimination and cyclopentadienyl ring hydrogenation

It has been shown that bis(cyclopentadienyl)(μ-cyclopentadiene)dinickel, (NiCP)₂(η-C₅H₆), and (η⁵-cyclopentadienyl) (η³-cyclopentenyl)nickel, CpNi(η³-C₅H₇), are formed in the reaction of nickelocene with methyl-lithium and with 1-phenyl-2-methyl-propenyl-lithium. The compound (NiCp)₂(μ-C₅H₆) can be only formed as a result of the reduction of the cyclopentadienyl ring bonded to the nickel atom whereas the formation of CpNi(η³-C₅H₇) can be explained by the further hydrogeneration of cyclopentadiene formed in the earlier reaction steps. (NiCp)₂(μ-C₅H₆) has been fully characterised spectrometrically and its X-ray structure determined. It crystallises in the orthorhombic system, space group Pnma, with four molecules per unit cell.     

Interaction of half-sandwich alkylmolybdenum(III) complexes with B(C6F5)3. The X-ray structure of [CpMo(η-C4H6)(μ-Cl)(μ-CH2)(O)MoCp][CH3B(C6F5)3]

The reactions of the half-sandwich molybdenum(III) complexes CpMo(η⁴-C₄H₄R₂)(CH₃)₂, where Cp=η⁵-C₅H₅ and R=H or CH₃, with equimolar amounts of B(C₆F₅)₃ have been investigated in toluene. EPR monitoring shows the formation of an addition product which does not readily react with Lewis bases such as ethylene, pyridine, or PMe₃. The analysis of the EPR properties and the X-ray structure of a decomposition product obtained from dichloromethane, [CpMo(η⁴-C₄H₆)(μ-Cl)(μ-CH₂)(O)MoCp][CH₃B(C₆F₅)₃], indicate that the borane attack has occurred at the methyl position.     

Noncentrosymmetric K2Mn3(SO4)3F2·4H2O and Rb2Mn3(SO4)3F2·2H2O with pseudo-KTP structures

Two fluoride sulfates,K₂Mn₃(SO₄)₃F₂·4H₂O(Ⅰ) and Rb₂Mn₃(SO₄)₃F₂·2H₂O (Ⅱ) are obtained by water solution method.Single-crystal X-ray diffraction analysis indicated that they crystallize in space groups of Cmc2_1.Their structures feature a pseudo-KTP structure consisting of interconnecting[Mn₃(SO₄)3F₂(H₂O)2]_∞ layers,which are further packing along the a axis with alkali metal cations balancing the charges.The structure relationships between the two compounds are discussed.Secondharmonic generation measurements manifest that Ⅰ and Ⅱ have similar second-harmonic generation responses of about 0.2 and 0.25 times that of KH₂PO₄.