Combined effects of headgroup charge and tail unsaturation of lipids on lateral organization and diffusion of lipids in model biomembranes

Lateral organization and dynamics of lipids in plasma membranes are crucial for several cellular processes such as signal transduction across the membrane and still remain elusive.In this paper,using coarse-grained molecular dynamics simulation,we theoretically study the combined effects of headgroup charge and tail unsaturation of lipids on the lateral organization and diffusion of lipids in ternary lipid bilayers.In neutral ternary lipid bilayers composed of saturated lipids,unsaturated lipids,and cholesterols,under the conditions of given temperature and components,the main factor for the phase separation is the unsaturation of unsaturated lipids and the bilayers can be separated into liquid-ordered domains enriched in saturated lipids and cholesterols and liquid-disordered domains enriched in unsaturated lipids.Once the headgroup charge is introduced,the electrostatic repulsion between the negatively charged lipid headgroups will increase the distance between the charged lipids.We find that the lateral organization and diffusion of the lipids in the(partially) charged ternary lipid bilayers are determined by the competition between the headgroup charge and the unsaturation of the unsaturated lipids.In the bilayers containing unsaturated lipids with lower unsaturation,the headgroup charge plays a crucial role in the lateral organization and diffusion of lipids.The headgroup charge may make the lipid domains unstable and even can suppress phase separation of the lipids in some systems.However,in the bilayers containing highly unsaturated lipids,the lateral organization and diffusion of lipids are mainly dominated by the unsaturation of the unsaturated lipids.This work may provide some theoretical insights into understanding the formation of nanosized domains and lateral diffusion of lipids in plasma membranes.     

Characterization of the Lipid Binding Pocket in GM2AP and SapB with EPR Spectroscopy

Electron paramagnetic resonance (EPR) spectroscopy of spin-labeled lipids in complex with the sphingolipid activator proteins, GM2AP and SapB, was utilized to characterize the hydrophobic binding pocket of these lipid transfer proteins. Specifically, the EPR line shapes reveal that the mobility of the labeled lipids within the binding pockets of the transfer proteins are more restricted than when in a lipid bilayer environment and that lipids in GM2AP are slightly more restricted than in SapB. EPR accessibility based relaxation measurements show that the relative ratios of oxygen and water accessibility to sites along the acyl chains in lipids in complex with GM2AP are similar to the profiles obtained for a lipid bilayer albeit with lowered values. The results for SapB are quite different, with the oxygen profile mimicking a lipid bilayer, but there is a higher degree of water accessibility to the acyl chains in the SapB complex, likely because of the location of the lipid at the dimer interface in SapB coupled to dynamics of the dimer.     

Lateral diffusion in equimolar mixtures of natural sphingomyelins with dioleoylphosphatidylcholine

Cellular membranes of mammals are composed of a complex assembly of diverse phospholipids. Sphingomyelin (SM) and phosphatidylcholine (PC) are important lipids of eukaryotic cellular membranes and neuronal tissues, and presumably participate in the formation of membrane domains, known as "rafts," through intermolecular interaction and lateral microphase decomposition. In these two-dimensional membrane systems, lateral diffusion of lipids is an essential dynamic factor, which might even be indicative of lipid phase separation process. Here, we used pulsed field gradient nuclear magnetic resonance to study lateral diffusion of lipid components in macroscopically oriented bilayers composed of equimolar mixtures of natural SMs of egg yolk, bovine brain, bovine milk and dipalmitoylphosphatidylcholine (DPPC) with dioleoylphosphatidylcholine (DOPC). In addition, differential scanning calorimetry was used as a complementary technique to characterize the phase state of the lipid bilayers. In fully liquid bilayers, the lateral diffusion coefficients in both DOPC/DPPC and DOPC/SM systems exhibit mean values of the pure bilayers. For DOPC/SM bilayer system, this behavior can be explained by a model where most SM molecules form short-lived lateral domains with preferential SM-SM interactions occurring within them. However, for bilayers in the presence of their low-temperature gel phase, lateral diffusion becomes complicated and cannot simply be understood solely by a simple change in the liquid phase decomposition.     

Influence of cholesterol on tetraphenylborate transport through rat brain lipid artificial membranes

Summary The influence of cholesterol on TΦB⁻ transport through rat brain lipid model membranes has been studied by performing conductance studies. As a model system was used a rat brain lipid filter supported membrane. The weight percent of cholesterol as referred to total lipids was varied in a wide range. Cholesterol produced an increase of the conductance of membranes induced by the lipophilic anion tetraphenylborate. Moroever, an increase in the cholesterol mole fraction had a strong influence on steady-state I–V characteristics of rat brain lipid model membranes. To speed up publication, the authors of this paper have agreed to not receive the proofs for correction.     

(1)H-MRS metabolic patterns for distinguishing between meningiomas and other brain tumors

Yet meningiomas have characteristic neuroimaging features, some other lesions are still confusing with meningiomas. The aim of this study was trying to find the typical (1)H-MRS metabolic factors of histologic subtyped meningiomas, schwannomas, metastases, and other brain tumors for differential diagnosis among them. (1)H-MRS using STEAM (TE/30 ms, TR/2 sec) and PRESS (TE/288 ms, TR/2 sec) sequences were performed on 44 untreated brain tumors. Obtained metabolic patterns from the typical spectra of meningioma, schwannoma, metastasis were compared with each other or other brain tumors to evaluate the usefulness for diagnosis between them. Alanine(Ala) was observed in 15 cases of the 19 meningiomas with a little variation to three histologic subtypes, while minimal lipids were observed in every 19 meningiomas. Elevated glutamate/glutamine(Glx) was detected in 12 cases of the meniningiomas. Increased myo-inositol(mI) was detected in 11 cases of the 13 schwannomas. Dominant lipids signals as well as long-T2 lipids were detected in every metastasis in conjunction with elevated choline (Cho). Enhanced Glx was observed in 4 cases of the 8 metastases without correlation of primary tumor site or types. Hemangiopericytoma showed different spectral patterns from typical meningiomas: only dominant Cho, minimal lipids and absence of Ala or Glx signals. These metabolic patterns in typical tumors may provide a basis for differential diagnosis (average value of chi(2) = 23.33, p < 0.01) between meningiomas and schwannomas as well as metastases. However proton spectral distinction among the different histologic subtypes of meningiomas was not definite.     

基于 1H NMR谱的给药赭石大鼠血清代谢组学研究

基于¹H NMR 的代谢组学方法对灌胃给药赭石的成年 Wistar 大鼠血清进行分析,运用主成分分析模式识别方法对所得数据进行处理,并对给药大鼠血清进行生化指标检测．研究结果表明,大鼠体内β-羟异丁酸、乙酸、丙酮、胆碱、甘油磷脂酰胆碱、葡萄糖、乳酸、低密度脂蛋白、极低密度脂蛋白和脂质等内源性代谢物浓度发生明显变化,可作为赭石的特征代谢物． 2 g/kg 和 5 g/kg 体重剂量赭石使大鼠机体产生大量活性氧化物(ROS),造成过氧化损伤,导致能量代谢和糖代谢紊乱,糖酵解反应增强,并且对肝功能造成了影响．
     

7 T 下脂肪对基于 NOE 的磁共振对比成像的影响

核 Overhauser 增强(NOE)效应在高场下的化学交换饱和转移中会造成 Z 谱高场位置的负背景信号,有望成为一种新的磁共振成像(MRI)对比机制．然而,研究指出, NOE信号的发生区域与主要的脂肪信号的频率位置有重叠,因此 MRI 中观察到的 NOE 信号有可能混合了脂肪信号．该文通过鸡蛋的模型实验,初步分析了脂肪含量较高的组织内脂肪信号对 NOE 效应的影响,并通过健康大鼠鼠脑及颅内肿瘤大鼠鼠脑实验,分析了脂肪对脑部 NOE 对比成像及基于 NOE 对比成像的疾病诊断的影响．结果表明,脂肪含量较高的组织内脂肪信号会引起伪 NOE 效应,并影响 NOE对比图像的准确性．
     

Hydrophobic characterization of intracellular lipids in situ by Nile Red red/yellow emission ratio

Nile Red (9-diethylamino-5H-benzo [alpha] phenoxazine-5-one) is a fluorescent lipophilic dye characterized by a shift of emission from red to yellow according to the degree of hydrophobicity of lipids. Polar lipids (i.e., phospholipids) which are mostly present in membranes, are stained in red whereas neutral lipids (esterified cholesterol and triglycerides) which are present in lipid droplets, are stained in yellow. Besides this marked, qualitative contrast between polar and neutral lipids, small differences of the hydrophobic strength could be assessed by the quantitative ratio of red and yellow emissions, in order to extend the discrimination of lipids within the groups of neutral and polar lipids. On the other hand, ratiometric data of red and yellow emissions have not yet been evaluated in the numerous previous light microscopy investigations which used Nile Red. In this work we show that the Nile Red red/yellow ratio enables discrimination of different lipids (monooleine>oleic acid>phosphatidylcholine>free cholesterol>trioleine>oleyl cholesteryl ester). We also show changes in the Nile Red red/yellow emission ratio of lipid droplets of 3T3 mouse fibroblasts induced by drugs interfering with the cholesterol cycle.     

2H NMR and polyelectrolyte-induced domains in lipid bilayers

2H NMR studies of polyelectrolyte-induced domain formation in lipid bilayer membranes are reviewed. The 2H NMR spectrum of choline-deuterated phosphatidylcholine (PC) reports on any and all sources of lipid bilayer surface charge, since these produce a conformation change in the choline head group of PC, manifest as a change in the 2H NMR quadrupolar splitting. In addition, homogeneous and inhomogeneous surface charge distributions are differentiated. Adding polyelectrolytes to lipid bilayers consisting of mixtures of oppositely charged and zwitterionic lipids produces 2H NMR spectra which are superpositions of two Pake sub-spectra: one corresponding to a polyelectrolyte-bound lipid population and the other to a polyelectrolyte-free lipid population. Quantitative analysis of the quadrupolar splittings and spectral intensities of the two sub-spectra indicate that the polyelectrolyte-bound populations is enriched with oppositely charged lipid, while the polyelectrolyte-free lipid population is correspondingly depleted. The same domain-segregation effect is produced whether cationic polyelectrolytes are added to anionic lipid bilayers or anionic polyelectrolytes are added to cationic lipid bilayers. The 2H NMR spectra permit a complete characterization of domain composition and size. The anion:cation ratio within the domains is always stoichiometric, as expected for a process driven by Coulombic interactions. The zwitterionic lipid content of the domains is always statistical, reflecting the systems tendency to minimize the entropic cost of demixing charged lipids into domains. Domain formation is observed even with rather short polyelectrolytes, suggesting that individual polyelectrolyte chains aggregate at the surface to form "superdomains". Overall, the polyelectrolyte bound at the lipid bilayer surface appears to lie flat along the surface and to be essentially immobilized through its multiple electrostatic contacts.     

MRI and MRS on preserved samples as a tool in fish ecology

Magnetic Resonance Imaging (MRI) and Magnetic Resonance Spectroscopy (MRS) gain increasing attention and importance as a tool in marine ecology. So far, studies were largely limited to morphological studies, e.g. for the creation of digital libraries. Here, the utility of MRI and MRS for ecologists is tested and exemplified using formalin preserved samples of the Antarctic silverfish, Pleuragramma antarctica. As this species lacks a swim bladder, buoyancy is attained by the deposition of large amounts of lipids that are mainly stored in subcutaneous and intermuscular lipid sacs. In this study MRI and MRS are not only used to study internal morphology, but additionally to investigate functional morphology and to measure parameters of high ecological interest. The data are compared with literature data obtained by means of traditional ecological methods.The results from this study show that MR scans are not only an alternative to histological sections (as shown before), but even allow the visualization of particular features in delicate soft tissues, such as Pleuragramma's lipid sacs. 3D rendering techniques proved to be a useful tool to study organ volumes and lipid content, which usually requires laborious chemical lipid extraction and analysis. Moreover, the application of MRS even allows for an analysis of lipids and fatty acids within lipid sacs, which wouldn't be possible using destructive methods. MRI and MRS, in particular when used in combination, have the capacity to provide useful data on parameters of high ecological relevance and thus have proven to be a highly valuable addition, if not alternative, to the classical methods.     

Coarse-grained simulations on interactions between spectrins and phase-separated lipid bilayers

Spectrin, the principal protein of the cytoskeleton of erythrocyte, plays a crucial role in the stability and flexibility of the plasma membrane of erythrocyte. In this work, we investigate the interactions between spectrins and phase-separated lipid bilayers using coarse-grained molecular dynamics simulation. We focus on the preference of spectrins with different lipids, the effects of the anionic lipids and the residue mutation on the interactions between spectrins and the lipid bilayers. The results indicate that spectrins prefer to contact with phosphatidylethanolamine (PE) lipids rather than with phosphatidylcholine (PC) lipids, and tend to contact with the liquid-disordered (Ld) domains enriched in unsaturated PE. Additionally, the anionic lipids, which show specific interaction with the positively charged or polar amino acids on the surface of the spectrins, can enhance the attraction between the spectrins and lipid domains. The mutation leads to the decrease of the structural stability of spectrins and increases the curvature of the lipid bilayer. This work provides some theoretical insights into understanding the erythrocyte structure and the mechanism of some blood diseases.     

How glucosylation triggers physical–chemical properties of curcumin: an experimental and theoretical study

In the present study, we investigate the structures of glucosylated curcumin derivatives with DFT at B3LYP/6‐31G* level. A conformational analysis is performed in order to determine the conformational minimum (GS) and rotational transition state (TS) of curcumin derivatives and then their electronic features are evaluated. HOMO and LUMO frontier orbitals and maps of electron density potential (MEPs) are plotted and compared. In order to correlate their predicted spectroscopic properties with IR, UV–vis and NMR experimental data we extended the theoretical study on electronic properties to different solvents (H₂O, MeOH, ACN, DMSO). The main finding is that the curcuminic core maintains the same geometrical and electronic structures in all compounds miming the metal coordination capability showed by curcumin. Therefore, we may confirm that the presence of glucose does not affect the electronic properties of the derivatives. Copyright © 2010 John Wiley & Sons, Ltd.     

含缩醛正离子类脂分子与蛋白质的相互作用

合成系列含缩醛的双链正离子类脂分子 ,并用荧光光谱研究其与牛血清蛋白 (BSA)的相互作用 .通过荧光的变化 ,解释蛋白质构象的变化 .在低类脂浓度时 ,少量类脂分子束缚在牛血清蛋白周围 ,荧光有很大幅度的淬灭 ,蛋白质本身肽链被解开 ,与此同时最大发射波长从 (3 44± 1)nm蓝移到 (3 3 1± 1)nm .由于疏水相互作用 ,更多类脂分子不断地聚集在蛋白质周围 ,牛血清蛋白中的两个色氨酸残基被完全地包裹在类脂分子形成的双分子膜中 ,荧光强度不断增加直到恒定不变     

Tracing Lysozyme-Lipid Interactions with Long-Wavelength Squaraine Dyes

The applicability of the two newly commercial available squaraine labels Square-670-NHS and Seta-635-NHS to exploring protein-lipid interactions has been evaluated. The labels were conjugated to lysozyme (Lz) (squaraine-lysozyme conjugates below referred to as Square-670-Lz and Seta-635-Lz), a structurally well-characterized small globular protein displaying the ability to interact both, electrostatically and hydrophobically with lipids. The lipid component of the model systems was represented by lipid vesicles composed of zwitterionic lipids egg yolk phosphatidylcholine (PC) and 1-stearoyl-2-oleoyl-sn-glycero-3-phosphocholine (SOPC), and their mixtures with anionic lipids either beef heart cardiolipin (CL) or 1-palmitoyl-2-oleoyl-sn-glycero-3-phosphoglycerol (POPG), respectively. Fluorescence intensity of Square-670-Lz was found to decrease upon association with lipid bilayer, while the fluorescence intensity of Seta-635-Lz displayed more complex behavior depending on lipid-to-protein molar ratio. Covalent coupling of squaraine labels to lysozyme exerts different influence on the properties of dye-protein conjugate. It was suggested that Square-670-NHS covalent attachment to Lz molecule enhances protein propensity for self-association, while squaraine label Seta-635-NHS is sensitive to different modes of lysozyme-lipid interactions—within the L:P range 6–11, when hydrophobic protein-lipid interactions are predominant, an aggregation of membrane-bound protein molecules takes place, thereby decreasing the fluorescence intensity of Seta-635-Lz. At higher L:P values (from 22 to 148) when electrostatic interactions are enhanced fluorescence intensity of Seta-635-Lz increases with increasing lipid concentrations.     

Ultra-stable temperature control in EPR experiments: thermodynamics of gel-to-liquid phase transition in spin-labeled phospholipid bilayers and bilayer perturbations by spin labels

An ultra-stable variable temperature accessory for EPR experiments with biological samples has been designed and tested. The accessory is comprised from a digitally controlled circulator bath that pumps fluid through high-efficiency aluminum radiators attached to an EPR resonator of a commercial X-band EPR spectrometer. Temperature stability of this new accessory after a 15 min re-equilibration is at least +/-0.007 K. For a standard 1-cm-long capillary sample arranged inside an EPR tube filled with silicon oil, the temperature variations do not exceed +/-0.033 K over the sample temperature range from 283 to 333 K. This new accessory has been tested by carrying out a comparative spin-labeling EPR and differential scanning calorimetry (DSC) study of the gel-to-liquid phase transition in multilamellar vesicles (MLV) composed of a synthetic phospholipid 1,2-dimyristoyl-sn-glycero-3-phosphatidylcholine (DMPC). We demonstrate that the gel-to-liquid phase transition temperatures of MLV DMPC measured by EPR and DSC agree within +/-0.02 K experimental error even though the sample for EPR study was labeled with 1 mol% of 5PC (1-palmitoyl-2-stearoyl-(5-doxyl)-sn-glycero-3 phosphocholine). Cooperative unit number measured by EPR, N=676+/-36, was almost 50% higher than that obtained from DSC (N=458+/-18). These high values of N indicate that (i) the lipid domains should include at least several spin-labeled lipid molecules and (ii) the spin-probe 5PC molecules are not excluded into domains that are different from the bulk lipid phase as was speculated earlier. Overall, our data provide DSC and EPR evidence that in studies of the gel-to-liquid phase transition, the effect of bilayer perturbation by spin-labeled lipids is negligible and therefore thermodynamic parameters of the phase transition can be accurately measured by spin-labeling EPR. This might serve as an indication when spin-labeled molecules with structures similar to those of lipids are introduced at low concentrations, they are easily accommodated by fluid phospholipid bilayers without significant losses of the lipid cooperativity.     

Influences of the Structure of Lipids on Thermal Stability of Lipid Membranes

The binding free energy (BFE) of lipid to lipid bilayer is a critical factor to determine the thermal or mechanical stability of the bilayer. Although the molecular structure of lipids has significant impacts on BFE of the lipid, there lacks a systematic study on this issue. In this paper we use coarse-grained molecular dynamics simulation to investigate this problem for several typical phospholipids. We find that both the tail length and tail unsaturation can significantly affect the BFE of lipids but in opposite way, namely, BFE decreases linearly with increasing length, but increases linearly with addition of unsaturated bonds. Inspired by the specific structure of cholesterol which is a crucial component of biomembrane, we also find that introduction of carbo-ring-like structures to the lipid tail or to the bilayer may greatly enhance the stability of the bilayer. Our simulation also shows that temperature can influence the bilayer stability and this effect can be significant when the bilayer undergoes phase transition. These results may be helpful to the design of liposome or other self-assembled lipid systems.     

Molecular dynamics simulations of the effects of sodium dodecyl sulfate on lipid bilayer

Molecular dynamics simulations have been performed on the fully hydrated lipid bilayer with different concentrations of sodium dodecyl sulfate(SDS). SDS can readily penetrate into the membrane. The insertion of SDS causes a decrease in the bilayer area and increases in the bilayer thickness and lipid tail order, when the fraction of SDS is less than 28%.Through calculating the binding energy, we confirm that the presence of SDS strengthens the interactions among the DPPC lipids, while SDS molecules act as intermedia. Both the strong hydrophilic interactions between sulfate and phosphocholine groups and the hydrophobic interactions between SDS and DPPC hydrocarbon chains contribute to the tight packing and ordered alignment of the lipids. These results are in good agreement with the experimental observations and provide atomic level information that complements the experiments.     

Insights into the molecular mechanism of membrane fusion from simulation: evidence for the association of splayed tails

We present coarse-grained simulations of fusion between two liposomes from which a detailed picture of lipid movements emerges. In these simulations the bilayers dilate at the contact edge, and the resulting increase in the area per lipid produces a tilting of the individual molecules as predicted. Fusion is initiated when some of these tilted lipids splay their aliphatic tails, such that the molecules are shared between the opposing leaflets. Multiple splayed lipids subsequently associate with their aliphatic tails in contact, which produces a new hydrophobic core. As the tails extend into a more parallel conformation the two outer leaflets become contiguous to produce a hemifused structure. The results have interesting implications for biological membrane fusion and suggest new possibilities for designing molecules that control fusion.     

Dietary lipid content influences the activity of lipogenic enzymes in the liver and on whole body delta13C values of Nile tilapia, Oreochromis niloticus (L.)

The use of stable isotope techniques for the reconstruction of diets has increased over the last decade. However, isotopic ratios in an animal are not only affected by the composition of the feed, but also by the amount of feed consumed. An uncertainty of up to 1 per thousand for both delta13C and delta15N values has been observed when the feeding level is unknown. This may have substantial effects on the results of back-calculation. As the feeding level of animals is unknown in nature, an additional indicator for their nutritional status is needed. High feeding levels and a consequent surfeit of dietary energy lead to the synthesis of lipids. In order to test whether the level of lipogenesis could be used as an indicator, Nile tilapia (Oreochromis niloticus) were fed four isonitrogenous and isoenergetic wheat-based semi-synthetic diets with different lipid contents (2.0 %, 4.5 %, 9.5 % and 13.3 %) for eight weeks. Body composition, gross energy content and delta13C values in the lipids and the lipid-free material were determined in diets and fish bodies. The livers of three fish per feeding group were assayed for the activity of two lipogenic enzymes, ATP-citrate lyase and malic enzyme. There was a strong negative correlation between delta13C values in the lipids of the individual fish and the apparent lipid conversion. The activities of lipogenic enzymes decreased with rising lipid content in the diet. The delta13C values in the lipids decreased significantly with increasing specific activity for both enzymes. In this experiment where lipogenesis was influenced by the composition of the diet, it was possible to determine the exact value for the trophic shift in relation to the enzyme activities. Further experiments to investigate the use of enzyme activities in situations where the feeding level of an animal is unknown are recommended.