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1.
Dolichols are isoprenologues which constitute an important component of biological membranes. However, an understanding of the effects of dolichols on the organization and dynamics of biological membranes has not been forthcoming. The experiments reported here are aimed at understanding the effects of dolichols on the physical properties of mouse brain synaptic plasma membranes. The effect of dolichols incorporated into mouse brain synaptic plasma membranes on fluorescent and electron spin resonance probes sensing the hydrophobic core differed from that of probes reporting closer to the surface of membrane bilayers. Dolichols significantly (P less than 0.01) lowered the polarization, limiting anisotropy, and order parameter of diphenylhexatriene in synaptic plasma membranes and liposomes extracted from synaptic plasma membranes, without changing the rotational relaxation time. Similarly, dolichol increased the fluidity reported by 16-doxylstearic acid in synaptic plasma membranes or liposomes extracted from synaptic plasma membranes. In contrast, dolichols exerted no effect on those properties for trans-parinaric acid or 5-doxylstearic acid in synaptic plasma membranes or liposomes derived therefrom. Dolichols can dramatically alter the structure and dynamics of lipid motion in synaptic plasma membranes and these effects are dependent on the location of the probe in the membrane. 相似文献
2.
Micrococcal nuclease digestion and light scattering are used to compare native chromatins with various histone H1[0] contents. The experimental data show that the higher the H1[0] content, the greater the ability to form compact structures with increasing ionic strength, and the lower the DNA accessibility to micrococcal nuclease. On the contrary, reconstituted samples from H1-depleted chromatin and pure individual H1 fractions behave in such a way that samples reconstituted with pure H1 degree give rise to a looser structure, more accessible to nuclease than samples reconstituted with H1-1. This contradiction suggests that the effect of H1o on chromatin structure must originate from the interaction of this histone with other components in native chromatin among which other histone H1 subfractions are good candidates. 相似文献
3.
W. Gibson Wood† Christine Gorka Friedhelm Schroeder‡ 《Journal of neurochemistry》1989,52(6):1925-1930
Alcohols, including ethanol, have a specific effect on transbilayer and lateral membrane domains. Recent evidence has shown that alcohols in vitro have a greater effect on fluidity of one leaflet as compared to the other. The present study examined effects of chronic ethanol consumption on fluidity of synaptic plasma membrane (SPM) exofacial and cytofacial leaflets using trinitrobenzenesulfonic acid (TNBS) labeling and differential polarized fluorometry of 1,6-diphenyl-1,3,5-hexatriene (DPH). Mice were administered ethanol or a control liquid diet for 3 weeks. Animals were killed and SPM prepared. The exofacial leaflet of SPM was significantly more fluid than the cytofacial leaflet in both groups, as indicated by limiting anisotropy of DPH. However, differences between the two leaflets were much smaller in the ethanol-treated group. Ethanol at concentrations seen clinically had a greater effect in vitro on the more fluid exofacial leaflet. This asymmetric effect of ethanol was significantly diminished in the exofacial leaflet of the ethanol-treated mice. Chronic ethanol consumption has a specific effect on membranes. Membrane functions that may be regulated by asymmetry of fluidity and lipid distribution may be altered by chronic ethanol consumption. 相似文献
4.
Rat liver chromatin was digested with micrococcal nuclease to various extents and fractionated into nucleosomes, di and trimers of nucleosomes on an isokinetic sucrose gradient. In conditions under which degradation of linker DNA within the particles was limited, the electrophoretic analysis of the histone content showed that the overall content of H1 histone increased from nucleosomes to higher order oligomers. Moreover, the histone H1 subfractions were found unevenly distributed among the chromatin subunits, one of them, H1--3 showing most variation. A more regular distribution of these subfractions was found in subunits obtained from a more extended digestion level of chromatin. It is suggested that the H1 subfractions differ in the protection they confer upon DNA. 相似文献
5.
Juan P. de-Torres David Blanco Ana B. Alcaide Luis M. Seijo Gorka Bastarrika María José Pajares Arrate Mu?oz-Barrutia Carlos Ortiz-de-Solorzano Ruben Pio Arantza Campo Usua Montes Victor Segura Jesús Pueyo Luis M. Montuenga Javier J. Zulueta 《PloS one》2013,8(4)
Rationale
Low-grade inflammation and emphysema have been shown to be associated with an increased risk of lung cancer. However, the systemic inflammatory response in patients with emphysema is still unknown.Objective
To compare the plasma cytokine profiles in two groups of current or former smokers without airway obstruction: a control group of individuals without computed tomography (CT) detected emphysema vs. a study group of individuals with CT detected emphysema.Methods
Subjects underwent a chest CT, spirometry, and determination of EGF, IL-15, IL-1ra, IL-8, MCP-1, MIP-1β, TGFα, TNFα, and VEGF levels in plasma. Cytokine levels in each group were compared adjusting for confounding factors.Results
160 current smokers and former smokers without airway obstruction participated in the study: 80 without emphysema and 80 subjects with emphysema. Adjusted group comparisons revealed significant reductions in EGF (−0.317, p = 0.01), IL-15 (−0.21, p = 0.01), IL-8 (−0.180, p = 0.02) and IL-1ra (−0.220, p = 0.03) in subjects with emphysema and normal spirometry.Conclusions
Current or former smokers expressing a well-defined disease characteristic such as emphysema, has a specific plasma cytokine profile. This includes a decrease of cytokines mainly implicated in activation of apoptosis or decrease of immunosurveillance. This information should be taken into account when evaluated patients with tobacco respiratory diseases. 相似文献6.
7.
Jorge Aramburu Raúl Antón Alejandro Rivas Juan Carlos Ramos Gorka S. Larraona Bruno Sangro 《Computer methods in biomechanics and biomedical engineering》2019,22(5):518-532
Balloon-occluded transarterial chemoembolisation (B-TACE) is an intraarterial transcatheter treatment for liver cancer. In B-TACE, an artery-occluding microballoon catheter occludes an artery and promotes collateral circulation for drug delivery to tumours. This paper presents a methodology for analysing the haemodynamics during B-TACE, by combining zero-dimensional and three-dimensional modelling tools. As a proof of concept, we apply the methodology to a patient-specific hepatic artery geometry and analyse two catheter locations. Results show that the blood flow redistribution can be predicted in this proof-of-concept study, suggesting that this approach could potentially be used to optimise catheter location. 相似文献
8.
Interaction of hagfish cathelicidin antimicrobial peptides with model lipid membranes 总被引:2,自引:0,他引:2
Hagfish intestinal antimicrobial peptides (HFIAPs) are a family of polycationic peptides exhibiting potent, broad-spectrum bactericidal activity. In an attempt to unravel the mechanism of action of HFIAPs, we have studied their interaction with model membranes. Synthetic HFIAPs selectively bound to liposomes mimicking bacterial membranes, and caused the release of vesicle-encapsulated fluorescent markers in a size-dependent manner. In planar lipid bilayer membranes, HFIAPs induced erratic current fluctuations and reduced membrane line tension according to a general theory for lipidic pores, suggesting that HFIAP pores contain lipid molecules. Consistent with this notion, lipid transbilayer redistribution accompanied HFIAP pore formation, and membrane monolayer curvature regulated HFIAP pore formation. Based on these studies, we propose that HFIAPs kill target cells, at least in part, by interacting with their plasma membrane to induce formation of lipid-containing pores. Such a membrane-permeabilizing function appears to be an evolutionarily conserved host-defense mechanism of antimicrobial peptides. 相似文献
9.
10.
Witold G. Szymanski Henrik Zauber Alexander Erban Michal Gorka Xu Na Wu Waltraud X. Schulze 《Molecular & cellular proteomics : MCP》2015,14(9):2493-2509
The plasma membrane is an important compartment that undergoes dynamic changes in composition upon external or internal stimuli. The dynamic subcompartmentation of proteins in ordered low-density (DRM) and disordered high-density (DSM) membrane phases is hypothesized to require interactions with cytoskeletal components. Here, we systematically analyzed the effects of actin or tubulin disruption on the distribution of proteins between membrane density phases. We used a proteomic screen to identify candidate proteins with altered submembrane location, followed by biochemical or cell biological characterization in Arabidopsis thaliana. We found that several proteins, such as plasma membrane ATPases, receptor kinases, or remorins resulted in a differential distribution between membrane density phases upon cytoskeletal disruption. Moreover, in most cases, contrasting effects were observed: Disruption of actin filaments largely led to a redistribution of proteins from DRM to DSM membrane fractions while disruption of tubulins resulted in general depletion of proteins from the membranes. We conclude that actin filaments are necessary for dynamic movement of proteins between different membrane phases and that microtubules are not necessarily important for formation of microdomains as such, but rather they may control the protein amount present in the membrane phases.Living cells need borders and molecular compartments for biochemical reactions and storage of metabolites. The plasma membrane therefore is a prerequisite for the evolution of different life forms. It consists of a phospholipid bilayer into which proteins and special lipid species such as sterols, sphingolipids, and glycolipids are inserted. The first complex model of plasma membrane was proposed in 1972 by Jonathan Singer and Garth Nicolson (1), replacing the concept of the plasma membrane as a strict protein–lipid–protein sandwich that was generally accepted until then. In Singer and Nicolson''s model, the cell membrane is a two-dimensionally oriented viscous solution in which the membrane constituents are orientated in the most thermodynamically favorable manner, hiding hydrophobic hydrocarbon chains inside the lipid bilayer and exposing polar and ionic groups to the aqueous phase. This fluid mosaic model also implied that membrane proteins as well as lipid components are distributed in a homogeneous lipid bilayer at long range, but they can form specific aggregates and phases at short range, which were also termed “lipid rafts” or membrane microdomains.Over the past 30 years, it has become evident that the plasma membrane is not such a homogeneous structure as it was initially proposed. We now know that the lipid bilayer is asymmetric (2) and that the free diffusion of membrane proteins is restricted by their interactions with intracellular and extracellular components (3). More recently, Simons and Ikonen suggested that large ordered phases, enriched with cholesterol and sphingolipids, emerge within the plasma membrane and that they function as platforms for enrichment of certain proteins while excluding others (4). This current membrane model suggests that the mixture of sterols and polar lipids within the plasma membrane can appear in two distinct phases: liquid disordered (Ld) and liquid ordered (Lo) phase (5). In this view, the so-called membrane microdomains are considered to be part of the Lo phase. Based on work on model membranes, it is suggested that lateral segregation of components into Ld and Lo phases occurs spontaneously (6) with the self-associating properties between sterols and highly saturated hydrocarbon chains of phopsho- and sphingolipids as the main driving force (7). Additionally, it is suggested that also specific lipid-protein and protein-protein interactions are essential for the formations of membrane domains as well as for stabilization of smaller nanodomains which subsequently may cause formation of larger platforms. In contrast to the animal cells, in plants these membrane microdomains seem to be rather immobile (8), possibly due to their attachment to the outer cell wall. More recently, it became obvious that membrane microdomains within a single cell are highly diverse and of different compositions (9). Generally, in the plant model, organisms'' plasma membrane microdomains turned out to be important in plant defense (10, 11), cell polarity (12, 13), and general signaling properties of the plasma membrane (14, 15).The cytoskeleton was identified as an essential cellular component with important roles in membrane topography, bordering, trafficking, and organelle movement (16). Single particle tracking in mammalian cells revealed that the transferrin receptor and macroglobulin receptor demonstrate normal Brownian diffusion but only within a specific membrane compartment (17). Two hypothetical models were proposed in order to explain this phenomenon (supplemental Fig. 1). Direct interactions between transmembrane proteins and cytoskeleton are suggested to creates a barrier, called “fence,” where cytosolic parts of transmembrane proteins collides with cytoskeletal components, limiting their diffusion to certain areas. These molecules can jump over the “fence” to a neighboring compartment, possibly due to the dynamic nature of the interaction of membrane proteins and cytoskeleton, where they are again temporally trapped (17). This phenomenon was recently described also in A. thaliana where the interplay between membrane microdomains and microtubules plays a role in secondary cell wall formation (reviewed in (18)). The second model assumes, additionally, that particular transmembrane proteins are anchored to and lined up along cytoskeleton and act as “pickets” to arrest free diffusion of other membrane components, including nontransmembrane proteins, within the enclosed compartment (19).For plants, the composition of these sterol-rich membranes phases was analyzed in several biochemical studies (14, 20–22). Thereby, low-density preparations of plasma membrane fractions after treatment with nonionic detergents (DRM1 fractions) were considered as a biochemical representation enriched in cellular membrane ordered phases or microdomains. Proteomic studies in mammalian cells consistently reported that the DRM fraction is highly enriched with several cytoskeletal proteins such as actin, tubulin, myosin, dynamin, actinin, and supervillin (23–25). Additionally, the level of phosphatidylinositol 4,5-bisphosphate (PI(4,5)P2), a lipid connecting the plasma membrane to actin filaments, was also significantly elevated in DRM preparations (26). Treatment with microtubule and actin depolymerizing agent results in drastic loss of many signaling proteins from these DRM fractions prepared from adult rat cardiac myocytes (27) or human embryonic retinal cells (28).Based on this knowledge, we propose two hypothetical models for the relationship between cytoskeleton and membrane microdomains for plant cells: (i) Actin filaments and microtubules could be important in the membrane phase separation or formation of the membrane microdomains themselves. In this case, disruption of the cytoskeleton would cause a lack of phase segregation in the plasma membrane. (ii) The cytoskeleton is only important for the incorporation of specific protein into the sterol-enriched regions but not for the general formation of these phase separations. This view implies that phase separations or membrane microdomains would still be present after cytoskeleton disruption but their protein composition can be different. Another possible scenario is (iii) that cytoskeletal elements serve as anchors for membrane microdomains at particular position in the plasma membrane, so the absence of these anchors would cause the increased mobility of microdomains (supplemental Fig. 1).The primary aim of this study was to characterize the interplay between cytoskeletal components and different membrane phases (microdomains) in A. thaliana suspension cell cultures. To reach this goal, biochemical and proteomic approaches were combined with confocal microscopy and activity assays measuring the influence of actin or tubulin disruption on the composition, localization, and biochemical properties of the sterol-enriched membrane microdomains. Thereby, for biochemical analyses, low-density detergent-resistant membrane fractions are analyzed as containing cellular sterol-rich membrane compartments. 相似文献