The effect of hydrostatic pressure on the bilayer structure of phosphatidylcholines containing omega-cyclohexyl fatty acyl chains

The barotropic behavior of aqueous dispersions of two representative omega-cyclohexyl phosphatidylcholines was investigated by pressure-tuning Fourier transform infrared spectroscopy. In the even-numbered homologue, 1,2-di-14-cyclohexyltetradecanoyl-sn-glycero-3-phosphocholine (14cyPC), the lipid molecules are orientationally disordered until the applied pressure reaches 2.1 kbar. This pressure marks the onset of correlation field splitting of the scissoring and rocking modes of the linear chain methylenes, as well as that of the cyclohexyl ring methylenes. It indicates immobilization of the entire acyl chains, whereby the zig-zag planes of the neighboring straight chain all-trans methylenes are oriented mainly perpendicular to each other. As judged from the magnitude of the correlation field splittings, the interchain interaction is weaker in 14cyPC than that in linear lipids (e.g., DMPC or DPPC). Upon an increase in pressure, up to 20 kbar, the zig-zag methylene planes in 14cyPC undergo a gradual transformation to a parallel orientation. In the odd-numbered homologue, 1,2-di-13-cyclohexyltridecanoyl-sn-glycero-3-phosphocholine (13cyPC), there is no correlation field splitting originating from the straight chain methylenes (up to 21 kbar). The linear, nonbranched segments of the omega-cyclohexyl chains in 13cyPC are closely packed with the all-trans methylene zig-zag planes oriented parallel to each other. There is, however, correlation field splitting of the ring methylenes, indicating interring interactions between the bulky cyclohexyl rings in opposing bilayer leaflets. There are major structural differences between the even- and odd-numbered homologues in the interfacial region, which remain even at high pressures. The ester carbonyl C = O stretching band in 14cyPC is a composite of two discrete bands which do not change considerably in intensity or frequency in the pressure range 2-20 kbar. In contrast, 13cyPC possesses an additional, low-frequency C = O stretching component at low pressures. As the pressure increases, the three component bands coalesce into a single C = O stretching band. Our results suggest equally oriented, fully hydrogen-bonded carbonyl groups in 13cyPC at pressures above approx. 10 kbar.     

Electrochemical and photon polarization modulation infrared reflection absorption spectroscopy study of the electric field driven transformations of a phospholipid bilayer supported at a gold electrode surface

Electrochemistry and polarization modulation Fourier transform infrared reflection absorption spectroscopy (PM-FTIRRAS) was employed to investigate fusion of small unilamellar vesicles of 1,2dioyl-sn-glycero-3-phosphatidyl choline (DOPC) onto the Au(111) electrode. Electrochemical studies demonstrated that the DOPC vesicles fuse and spread onto the gold electrode surface at small charge densities -8 microC cm(-2)相似文献   

Formation of oriented polypeptides on Au(111) surface depends on the secondary structure controlled by peptide length.

We synthesized three different lengths of poly(L-lysine) containing an -SH group at the terminal (PLL(n)-SH, n (polymerization degree) = 4, 10, 30) and adsorbed them on an Au(111) surface. To analyze the formation process and the structure of self-assembled monolayers (SAMs), we used atomic force microscopy (AFM) and Fourier transform infrared reflection absorption spectra (FT-IR RAS). At the initial stage of SAM growth, formation of nanosize domains was confirmed by AFM imaging. The alpha-helical PLL(30)-SH exhibited a well-defined SAM structure after adsorption reached equilibrium. The alpha-helical PLL(30)-SH was almost perpendicular to the gold surface and exhibited interesting molecular packing due to the secondary structure of PLL(30)-SH and the underlying Au(111) array. The tilt angle of the helix axis from the substrate normal was estimated to be about 50 degrees (AFM) and 44 degrees (FT-IR RAS) respectively. On the other hand, PLL(4)-SH and PLL(10)-SH formed beta-sheet-type SAMs on the Au(111) surface based on the structure determined by FT-IR RAS spectrum.     

The effect of surface curvature on the head-group structure and phase transition properties of phospholipid bilayer vesicles

Proton nuclear magnetic resonance spectra at 360 MHz of small sonicated distearoyl phosphatidylcholine vesicles show easily distinguishable resonances due to choline N-methyl head-group protons located in the inner and outer bilayer halves. A study of the chemical shift of these resonances as a function of temperature reveals that the splitting between them increases below the phase transition. This occurs as a result of an upfield shift of the inner layer resonance at the phase transition. Consideration of the possible causes of this effect results in the conclusion that, at the phase transition, there is a change in the organization of the inner layer head-groups which does not occur for the outer layer head-groups.     

Effect of surface coverage of gold(111) electrode with cysteine on the chiral discrimination of DOPA

The enantioselectivity imparted to a gold electrode by modifying its surface with a self-assembled monolayer (SAM) of cysteine (Cys) was investigated for the electrochemical redox reaction of 3,4-dihydroxyphenylalanine (DOPA). A cyclic voltammetric study of the redox reaction revealed that the enantioselectivity was determined by the surface coverage of the gold electrode with Cys molecules. The electrode modified with approximately 1.8 x 10(14) Cys molecules cm(-2) exhibited enantioselectivity in the voltammogram for the oxidation and reduction of DOPA, while the voltammograms obtained by the electrodes with either more or less surface coverages did not exhibit significant enantioselectivity. It is suggested that the accessibility of DOPA to that area of the gold surface which is not blocked by Cys molecules at an optimum surface coverage, is required for the enantioselective redox reaction of DOPA to proceed.     

Synthesis, solid-state structure, and surface properties of end-capped poly(L-lactide)

End-capped poly(L-lactide) (PLLA) samples with dodecyl or 2-(2-(2-methoxyethoxy)ethoxy)ethyl (MEEE) ester were synthesized by ring-opening polymerization of L-lactide in the presence of zinc dodecanoxide or zinc 2-(2-(2-methoxyethoxy)ethoxy)ethoxide as a catalyst, respectively. On the basis of NMR analysis, it was confirmed that the carboxylic acid chain ends of PLLA molecules were selectively substituted by dodecyl or MEEE ester groups. To evaluate the wettability on the surface of end-capped PLLA films, the advancing contact angle (thetaa) with water was measured. The amorphous PLLA films showed relatively similar thetaa values regardless of the chemical structure of the polymer chain end. In contrast, the thetaa values of semicrystalline films were varied over a wide range, dependent on the chemical structure of the chain end. In addition, the thetaa values of dodecyl ester end-capped PLLA film with low molecular weight increased with an increase in the crystallization temperature. Both the crystallinity and lamellar thickness of dodecyl ester end-capped PLLA films increased with the crystallization temperature. These results suggest that the segregation of the chain ends on the PLLA film surface was strongly affected by the crystallization conditions.     

Primary structure of the glycan chains of normal C 1 esterase inhibitor (C 1-INH) after NMR analysis at 400 MHz

The primary structural analysis of O- and N-linked carbohydrate chains of the C-1-esterase inhibitor purified from normal serum was carried out by 400-MHz 1H-NMR spectroscopy. C-1-esterase inhibitor protein of a molecular weight of 116,000 daltons contains 24 O-glycans: NeuAc (alpha 2-3) Gal (beta 1-3) GalNAc, 4 N-glycans: NeuAc (alpha 2-6) Gal (beta 1-4) (GlcNAc (beta 1-2) Man (alpha 1-3) [NeuAc (alpha 2-6) Gal (beta 1-4) GlcNAc (beta 1-2) Man (alpha 1-6)] Man (beta 1-4) GlcNAc (beta 1-4) GlcNAc and 2 N-glycans: NeuAc (alpha 2-3) Gal (beta 1-4) GlcNAc (beta 1-2) Man (alpha 1-3) [NeuAc (alpha 2-3) Gal (beta 1-4) GlcNAc (beta 1-2) Man (alpha 1-6)] Man (beta 1-4) GlcNAc (beta 1-4) GlcNAc. 30% of the N-glycans are fucosylated.     

Synthesis, structure, magnetic and electrochemical properties of an oxydiacetate iron(II) complex

Compound [{Fe(oda)(H₂O)₂} · H₂O]_n (1) [oda=O(CH₂COO)₂ ²⁻] has been obtained by reaction of FeCl₂ · 4H₂O with a 1:1 mixture of O(CH₂COOH)₂ and Na₂CO₃ in water. The structure is polymeric with concatenated {Fe(oda)(H₂O)₂} units extended in one direction. Each iron centre is six-coordinated by the tridentate planar oda ligand, two mutually trans water molecules and one oda oxygen atom from an adjacent {Fe(oda)(H₂O)₂} unit. Complex 1 is isomorphous with cobalt and zinc analogues. Magnetic susceptibility measurements down to 2 K showed high-spin non-correlated Fe(II) ions with a typical Curie-Weiss behaviour. Differential-pulse voltammetry establishes a value of 0.488 V versus NHE for the Fe(III)/Fe(II) redox potential of this iron complex in 0.25 mol dm⁻³ NaNO₃.     

Spectroscopic studies on the interaction of Au(III) with nucleosides,nucleotides, and dimethyl phosphate

A series of complexes of Au(III) with nucleosides and nucleotides and their methyl derivatives in different stoichiometry have been prepared. Ultraviolet, visible, ir, and nmr studies have been performed to determine the site of binding of these ligands with the metal ion. In (1:4) Au(III): guanosine complex, N₇ is the binding site, whereas at 1:1 complex, a bidentate type of chelation through C₆O and N₇ is observed. C₆-NH₂ is favored over N₁ as coordinating site at all stoichiometry in the adenosine complex. Inosine binds through N₁ at r = 1. In cytidine, N₁ is the binding site, whereas thymidine reacts only at high pH. In the case of nucleotides a bidentate type of chelation through the phosphate and the ring nitrogen occurs. The phosphate binding ability of Au(III) was further confirmed by studying the interaction of Au(III) with dimethyl phosphate—a conformational analog of the phosphate backbone in DNA chain.     

Aggregation properties of semisynthetic GM1 ganglioside (II3Neu5AcGgOse4Cer) containing an acetyl group as acyl moiety

The aggregative properties of GM1 ganglioside containing an acetyl group as acyl moiety [GM1(acetyl)] in aqueous solution have been studied by static and dynamic light scattering measurements and surface tension experiments. GM1 (acetyl) spontaneously aggregates as small micelles showing a hydrodynamic radius and molecular weight of 34 A and 102 kDa, respectively, down to a concentration of 2.0 x 10(-5) M.     

Crystal structure at 2.8 A of Huntingtin-interacting protein 1 (HIP1) coiled-coil domain reveals a charged surface suitable for HIP1 protein interactor (HIPPI)

Huntington's disease is a genetic neurological disorder that is triggered by the dissociation of the huntingtin protein (htt) from its obligate interaction partner Huntingtin-interacting protein 1 (HIP1). The release of the huntingtin protein permits HIP1 protein interactor (HIPPI) to bind to its recognition site on HIP1 to form a HIPPI/HIP1 complex that recruits procaspase-8 to begin the process of apoptosis. The interaction module between HIPPI and HIP1 was predicted to resemble a death-effector domain. Our 2.8-Å crystal structure of the HIP1 371-481 subfragment that includes F432 and K474, which is important for HIPPI binding, is not a death-effector domain but is a partially opened coiled coil. The HIP1 371-481 model reveals a basic surface that we hypothesize to be suitable for binding HIPPI. There is an opened region next to the putative HIPPI site that is highly negatively charged. The acidic residues in this region are highly conserved in HIP1 and a related protein, HIP1R, from different organisms but are not conserved in the yeast homologue of HIP1, sla2p. We have modeled ∼ 85% of the coiled-coil domain by joining our new HIP1 371-481 structure to the HIP1 482-586 model (Protein Data Bank code: 2NO2). Finally, the middle of this coiled-coil domain may be intrinsically flexible and suggests a new interaction model where HIPPI binds to a U-shaped HIP1 molecule.     

Synthesis, structure and cytotoxicity studies of four-coordinate bis (cis-bis(diphenylphosphino)ethene) gold(I) complexes, [Au(dppey)2]X

Four-coordinate 1:2 gold(I) complex salts with cis-bis(diphenylphosphino)ethene, [Au(dppey)₂]X have been synthesized for X = PF₆⁻, CF₃SO₃⁻, BF₄⁻, Cl⁻, Br⁻ and BPh₄⁻ and characterized by NMR spectroscopy and electrospray mass spectrometry. Single crystal X-ray structure determinations show the BF₄⁻, Cl⁻ and Br⁻ complexes to be isostructural, although with different degrees of hydration, while the BPh₄⁻ complex crystallizes as an acetone solvate with two molecules in the asymmetric unit. The Au(P-P)₂ core for the BF₄⁻, Cl⁻ and Br⁻ complexes adopts D₂ symmetry with Au-P bond lengths 2.3980(7)-2.4009(7) Å and inter-ligand P-Au-P angles 114.78(2)-127.82(2))°. The Au(P-P)₂ core in the BPh₄⁻ complex is unsymmetrical with Au-P bond lengths 2.364(1)-2.420(1) Å and inter-ligand P-Au-P angles 104.76(5)-137.50(4)°. In vitro cytotoxicity studies show the PF₆⁻, CF₃SO₃⁻, BF₄⁻, Cl⁻, Br⁻ and I⁻ complexes to be potent and selective growth inhibitors of the human cell lines MCF7 (hormone-dependent breast cancer), MDA-MB-231 (hormone-independent breast cancer), MM96L (melanoma), CI80-13S (cisplatin resistant ovarian cancer) and a normal cell line NFF (neonatal foreskin fibroblasts), achieving IC₅₀ values between 13 and 196 nM. The halogen and triflate salts were approximately twice as potent towards the MCF7 and MDA-MB-231 cell lines compared to the PF₆⁻ and BF₄⁻ derivatives; while the cytotoxicity of all complexes towards the sensitive CI80-13S and MM96L cancer cell lines was approximately 10-fold greater than that displayed towards the normal human cell line (NFF).     

Structural recognition of an ICAM-1 peptide by its receptor on the surface of T cells: conformational studies of cyclo (1, 12)-Pen-Pro-Arg-Gly-Gly-Ser-Val-Leu-Val-Thr-Gly-Cys-OH.

The purpose of this study is to elucidate the solution conformation of cyclic peptide 1 (cIBR), cyclo (1, 12)-Pen1-Pro2-Arg3-Gly4-Gly5-Ser6-Val7-Leu8-V al9-Thr10-Gly11-Cys12-OH, using NMR, circular dichroism (CD) and molecular dynamics (MD) simulation experiments. cIBR peptide (1), which is derived from the sequence of intercellular adhesion molecule-1 (ICAM-1, CD54), inhibits homotypic T-cell adhesion in vitro. The peptide hinders T-cell adhesion by inhibiting the leukocyte function-associated antigen-1 (LFA-1, CD11a/CD18) interaction with ICAM-1. Furthermore, Molt-3 T cells bind and internalize this peptide via cell surface receptors such as LFA-1. Peptide internalization by the LFA-1 receptor is one possible mechanism of inhibition of T-cell adhesion. The recognition of the peptide by LFA-1 is due to its sequence and conformation; therefore, this study can provide a better understanding for the conformational requirement of peptide-receptor interactions. The solution structure of 1 was determined using NMR, CD and MD simulation in aqueous solution. NMR showed a major and a minor conformer due to the presence of cis/trans isomerization at the X-Pro peptide bond. Because the contribution of the minor conformer is very small, this work is focused only on the major conformer. In solution, the major conformer shows a trans-configuration at the Pen1-Pro2 peptide bond as determined by HMQC NMR. The major conformer shows possible beta-turns at Pro2-Arg3-Gly4-Gly5, Gly5-Ser6-Val7-Leu8, and Val9-Thr10-Gly11-Cys12. The first beta-turn is supported by the ROE connectivities between the NH of Gly4 and the NH of Gly5. The connectivities between the NH of Ser6 and the NH of Val7, followed by the interaction between the amide protons of Val7 and Leu8, support the presence of the second beta-turn. Furthermore, the presence of a beta-turn at Val9-Thr10-Gly11-Cys12 is supported by the NH-NH connectivities between Thr10 and Gly11 and between Gly11 and Cys12. The propensity to form a type I beta-turn structure is also supported by CD spectral analysis. The cIBR peptide (1) shows structural similarity at residues Pro2 to Val7 with the same sequence in the X-ray structure of D1-domain of ICAM-1. The conformation of Pro2 to Val7 in this peptide may be important for its binding selectivity to the LFA-1 receptor.     

The preparation,spectral studies,and the crystal structure of dimethylbis(O-ethylxanthato)tin(IV)

Tin-119 NMR data indicate that the tin atom in (CH₃)₂Sn(S₂COC₂H₅)₂ is four co-ordinated in dichloromethane solution. However, single crystal X-ray analysis shows the tin atom to be six co-ordinated in the solid state in which the bidentate xanthate ligands display gross asymmetry in their mode of co-ordination to the tin. The crystals are molecular and there is no association between neighbouring molecules. The unit cell of Me₂Sn(exa)₂ is orthorhombic, Pnma, a = 14.165(1), b = 7.675(9), c = 13.977(2) Å with Z = 4. The structure was refined by conventional least squares methods with final R 0.041 and R_w 0.043 for 1229 unique reflections with 1 ? 2σ(I).     

Ganglioside GM(1) biphasically regulates the activity of protein kinase C by the effects on the structure of the lipid bilayer

Addition of a small amount of ganglioside GM(1) to phosphatidylserine (PS) liposomes, a gradual increase of protein kinase C (PKC) activity was recorded up to about 2 mol% GM(1) where the maximal enzyme activity was obtained. Then the activity of PKC began to decline and even turned to be inhibited with the further increase of GM(1) content. It was also indicated that GM(1)/PS binary liposomes had the highest membrane fluidity and very low spatial density of lipid headgroups which was demonstrated in the MC-540 studies due to the interposition of GM(1) when the liposomes contained about 2 mol% GM(1). Besides, the liposomes containing about 2 mol% GM(1) provided a more hydrophobic environment for PKC than the liposomes containing less or more GM(1) which was indicated in the Acrylodan experiments. These factors commonly induced PKC to be stimulated maximally. Whether at the lower or higher GM(1) content, the membrane structure was not the most suitable to support the activity of PKC, which declined as a consequence.     

Fourier transform infrared studies of secondary structure and orientation of pulmonary surfactant SP-C and its effect on the dynamic surface properties of phospholipids

SP-C, a highly hydrophobic, 3.7-kDa protein constituent of lung surfactant, has been isolated from bovine lung lavage, purified, and reconstituted into binary lipid mixtures of 1,2-dipalmitoyl-phosphatidylcholine (DPPC) and 1,2-dipalmitoylphosphatidylglycerol (DPPG). Fourier transform infrared (FT-IR) spectroscopy has been applied to examine SP-C secondary structure, the average orientation of alpha-helical segments relative to the bilayer normal in membrane films, and the effect of protein on the thermotropic properties of the phospholipid acyl chains. In addition, dynamic surface measurements were made on phospholipid films at the A/W interface in the presence and absence of SP-C. SP-C (0.5 mol %) was found to possess about 60% alpha-helical secondary structure in lipid vesicles. Higher levels (1.5 mol %) of SP-C resulted in a slight increase of beta-forms, possibly resulting from protein aggregation. The helical segments exhibited an average angle of orientation of about 24 degrees with respect to the bilayer normal, suggesting a trans-bilayer orientation of the peptide. The observation that 70% of the peptide bond hydrogens are hard to exchange in D2O further reflects the hydrophobic nature of the molecule. SP-C produced little effect on the thermotropic properties of the binary lipid mixture, as measured from acyl chain C-H and C-D stretching frequencies. However, the presence of 1 mol % protein markedly reduced the viscance and increased the elasticity of surface films suggesting a mechanism by which SP-C facilitates the spreading of phospholipids on an aqueous surface. The possible physiological consequences of these observations are discussed.     

Genomic structure and chromosome mapping of the genes encoding clathrin-associated adaptor medium chains mu1A (Ap1m1) and mu1B (Ap1m2)

The protein mu1B is a member of the medium chain family of the clathrin-associated adaptor complex and is expressed exclusively in epithelial cells. We determined the genomic structure of previously cloned murine genes for mu1B (Ap1m2) and its closely related homolog, mu1A (Ap1m1). Comparison of their genomic structures revealed that the positions of introns are identical between these two genes, except for the insertion of an additional intron in Ap1m1 (intron 4). By contrast, these structures are different from that of the more distantly related Ap2m1 gene encoding mu2. Taken together with the similarity of amino acid sequences among these genes, the data presented in this study suggest that Ap1m1/2 and Ap2m1 diverged long before the separation of Ap1m1 and Ap1m2, which most likely resulted from a relatively recent gene duplication. We also mapped AP1M2 to human chromosome 19p13.2 and Ap1m2 to the proximal region of mouse chromosome 9. The results are consistent with the fact that these regions are syntenic.     

Hydrido iridium(III) complexes of azoaromatic ligands. Isolation, structure and studies of their physicochemical properties

Reactions of 2-(arylazo)pyridine (L^a-c) with [IrCl₃(PPh₃)₂] in two different solvents, viz. ethanol and toluene are reported. In refluxing toluene two new isomeric (mer and fac geometries) iridium complexes, having molecular formula [IrCl₃(PPh₃)(L)] (1 and 2) have been isolated. The reaction in refluxing ethanol yielded two new hydrido complexes of molecular formula [IrHCl₂(PPh₃)(L)] (3) and [IrHCl(PPh₃)₂(L)]Cl (4) along with the compound 2. All the complexes have been thoroughly characterized by NMR, UV-Vis spectroscopy, cyclic voltammetry and X-ray crystallographic analysis. The ¹H NMR spectra of the hydrido complexes 3 and 4 showed a doublet and a triplet signals at δ −20.43 and −14.82 respectively due to coupling with magnetically equivalent phosphorous nuclei. Strong trans influence of the π-acceptor ligands guided the X-ray structural parameters; bonds trans to the these ligands are unusually long. Similar elongation effect was also noted for the bonds trans to the coordinated hydrido ligand. UV-Vis-NIR spectrum consisted of multiple transitions in the UV and visible regions. Cyclic voltammetry of each of these complexes has exhibited a reductive response between −0.25 and −0.55 V, which has been assigned to azo-ligand reduction. The compound 3, however, showed a quasireversible oxidative wave near 1.45 V, due to Ir^III/Ir^IV couple.