Regulation Of Lipid Composition Research Articles

Lipid flip-flop vs. lateral diffusion in the relaxation of hemifusion diaphragms

Molecular dynamics simulations of a solvent-free coarse-grained lipid model are used to characterize the mechanisms by which lipid-bilayer hemifusion diaphragm (HD) intermediates relax, across a range of global compositions of negative intrinsic curvature (NIC) lipids and neutral-curvature lipids. At low concentrations of NIC lipids, rapid fission produces a double bilayer end state through a lateral diffusion-based mechanism enabled by spontaneous rim-pore defects. At moderately higher NIC lipid concentrations, rim pores are absent and stable leaflet three-junctions persist, revealing an HD relaxation mechanism entirely reliant on lipid flip-flop, and end states that are either stable fusion pores or stable HD's. These fusogenic systems exhibit dynamics highly dependent on NIC lipid concentration via an underlying sensitivity of flip-flop rates for neutral lipids on NIC lipid concentration. This work illustrates that HD dynamics may be altered through regulation of lipid composition in the immediate three-junction region. This work further highlights the potential role of flippases in biological fusion and the importance of lipid composition on fusion dynamics.

Remodeling of the postsynaptic plasma membrane during neural development.

Neuronal synapses are the fundamental units of neural signal transduction and must maintain exquisite signal fidelity while also accommodating the plasticity that underlies learning and development. To achieve these goals, the molecular composition and spatial organization of synaptic terminals must be tightly regulated; however, little is known about the regulation of lipid composition and organization in synaptic membranes. Here we quantify the comprehensive lipidome of rat synaptic membranes during postnatal development and observe dramatic developmental lipidomic remodeling during the first 60 postnatal days, including progressive accumulation of cholesterol, plasmalogens, and sphingolipids. Further analysis of membranes associated with isolated postsynaptic densities (PSDs) suggests the PSD-associated postsynaptic plasma membrane (PSD-PM) as one specific location of synaptic remodeling. We analyze the biophysical consequences of developmental remodeling in reconstituted synaptic membranes and observe remarkably stable microdomains, with the stability of domains increasing with developmental age. We rationalize the developmental accumulation of microdomain-forming lipids in synapses by proposing a mechanism by which palmitoylation of the immobilized scaffold protein PSD-95 nucleates domains at the postsynaptic plasma membrane. These results reveal developmental changes in lipid composition and palmitoylation that facilitate the formation of postsynaptic membrane microdomains, which may serve key roles in the function of the neuronal synapse.

Membrane-Mediated Regulation of the Intrinsically Disordered CD3ϵ Cytoplasmic Tail of the TCR

The regulation of T-cell-mediated immune responses depends on the phosphorylation of immunoreceptor tyrosine-based activation motifs (ITAMs) on T-cell receptors. Although many details of the signaling cascades are well understood, the initial mechanism and regulation of ITAM phosphorylation remains unknown. We used molecular dynamics simulations to study the influence of different compositions of lipid bilayers on the membrane association of the CD3ϵ cytoplasmic tails of the T-cell receptors. Our results show that binding of CD3ϵ to membranes is modulated by both the presence of negatively charged lipids and the lipid order of the membrane. Free-energy calculations reveal that the protein-membrane interaction is favored by the presence of nearby basic residues and the ITAM tyrosines. Phosphorylation minimizes membrane association, rendering the ITAM motif more accessible to binding partners. In systems mimicking biological membranes, the CD3ϵ chain localization is modulated by different facilitator lipids (e.g., gangliosides or phosphoinositols), revealing a plausible regulatory effect on activation through the regulation of lipid composition in cell membranes.

Sterol regulatory element binding protein-dependent regulation of lipid synthesis supports cell survival and tumor growth

BackgroundRegulation of lipid metabolism via activation of sterol regulatory element binding proteins (SREBPs) has emerged as an important function of the Akt/mTORC1 signaling axis. Although the contribution of dysregulated Akt/mTORC1 signaling to cancer has been investigated extensively and altered lipid metabolism is observed in many tumors, the exact role of SREBPs in the control of biosynthetic processes required for Akt-dependent cell growth and their contribution to tumorigenesis remains unclear.ResultsWe first investigated the effects of loss of SREBP function in non-transformed cells. Combined ablation of SREBP1 and SREBP2 by siRNA-mediated gene silencing or chemical inhibition of SREBP activation induced endoplasmic reticulum (ER)-stress and engaged the unfolded protein response (UPR) pathway, specifically under lipoprotein-deplete conditions in human retinal pigment epithelial cells. Induction of ER-stress led to inhibition of protein synthesis through increased phosphorylation of eIF2α. This demonstrates for the first time the importance of SREBP in the coordination of lipid and protein biosynthesis, two processes that are essential for cell growth and proliferation. SREBP ablation caused major changes in lipid composition characterized by a loss of mono- and poly-unsaturated lipids and induced accumulation of reactive oxygen species (ROS) and apoptosis. Alterations in lipid composition and increased ROS levels, rather than overall changes to lipid synthesis rate, were required for ER-stress induction.Next, we analyzed the effect of SREBP ablation in a panel of cancer cell lines. Importantly, induction of apoptosis following SREBP depletion was restricted to lipoprotein-deplete conditions. U87 glioblastoma cells were highly susceptible to silencing of either SREBP isoform, and apoptosis induced by SREBP1 depletion in these cells was rescued by antioxidants or by restoring the levels of mono-unsaturated fatty acids. Moreover, silencing of SREBP1 induced ER-stress in U87 cells in lipoprotein-deplete conditions and prevented tumor growth in a xenograft model.ConclusionsTaken together, these results demonstrate that regulation of lipid composition by SREBP is essential to maintain the balance between protein and lipid biosynthesis downstream of Akt and to prevent resultant ER-stress and cell death. Regulation of lipid metabolism by the Akt/mTORC1 signaling axis is required for the growth and survival of cancer cells.

Molecular Convergence of Bacterial and Eukaryotic Surface Order

The conservation of fluidity is a theme common to all cell membranes. In this study, an analysis of lipid packing was conducted via C-laurdan spectroscopy of cell surface membranes prepared from representative species of Bacteria and Eukarya. We found that despite their radical differences in composition (namely the presence and absence of membrane-rigidifying sterol) the membrane order of all taxa converges on a remarkably similar level. To understand how this similarity is constructed, we reconstituted membranes with either bacterial or eukaryotic components. We found that transmembrane segments of proteins have an important role in buffering lipid-mediated packing. This buffering ensures that sterol-free and sterol-containing membranes exhibit similar barrier properties.

Molecular Dynamics Simulations of Monolayers and Membranes with Phosphatidylinositol Bisphosphate

Proper functionality of biological membranes depends on the regulation of lipid composition and localization. Spatial localization of molecules within the lipid bilayer depends on both steric effects due to their acyl chains and attractive or repulsive interactions between lipid headgroups, such as those mediated by the electrostatic charge of the lipid. Most eukaryotic lipids are zwitterionic or have a charge of −1 at physiological pH, but some lipids such as phosphatidylinositol bisphosphate (PtdInsP2) bear a net charge of −4. The ability of these highly charged lipids to interact with monovalent and divalent cations affects their spatial organization and temporal distribution on the cytoplasmic side of membranes. In turn, these lipids act as important effectors of apoptosis, inflammation, motility, and proliferation through their interactions with proteins at the membrane interface and transmembrane ion channels. In particular, PtdInsP2 is able to stabilize the open conformation of both eukaryotic potassium channels and those produced by bacteria, which normally lack PtdInsP2 lipids in their membrane. We hypothesize that in some settings PtdInsP2 acts by changing the physical-chemical properties of the membrane rather than by specific biochemical binding to proteins. To explore this hypothesis using molecular dynamics simulations, we completed a set of novel parameterizations at the MP2/6-31+G∗ level of theory for the three PtdInsP2 isoforms in several geometries and protonation states. Here we report that there are key differences in the way calcium and magnesium interact with the PtdInsP2 headgroup and that divalent cations likely influence the propensity of the lipids to form clusters.

Paraoxonase 1 R/Q alleles are associated with differential accumulation of saturated versus 20:5n3 fatty acid in human adipose tissue

Serum paraoxonase 1 (PON1) function has been associated with human cardiovascular disease. The projected mechanism postulates interaction of PON1 with lipoproteins and insulin signaling resulting in alterations in lipid homeostasis. Recently, PON2 was shown to directly regulate triglyceride accumulation in macrophages and PON1 was detected in the interstitial space of adipocytes. The aims of the present study were a) to examine the relationship of the PON1 function with serum parameters related to lipid homeostasis, and b) to examine a possible role of PON1 in the regulation of lipid composition in the human adipose tissue. Two important genetic variations with functional impact on PON1 activity in humans are the Q192R and the L55M. The present study evaluated the impact of the Q192R and the L55M polymorphisms in a cross-section of the population on the island of Crete, as regards to PON1 activity, plasma lipids/lipoproteins, parameters of the metabolic syndrome, and the fatty acid composition of the adipose tissue. We detected a significant association of the polymorphisms with blood pressure, fasting blood glucose, triglycerides, apolipoprotein B, serum iron, and homocysteine. Furthermore, a novel function is suggested for PON1 on the fatty acid composition in the adipose tissue through the positive association of the R allele with saturated fatty acid and of the Q allele with 20:5n3 fatty acid deposition.

Regulation of lipid composition in biological membranes—biophysical studies of lipids and lipid synthesizing enzymes

The study of the role played by membrane lipids, functional lipidomics, has become increasingly important in membrane biology. The physico-chemical properties of the lipids in biological membranes are subject to some fundamental requirements. In general, the acyl chains shall be in a liquid-like state to keep the membrane proteins active, and the lipids must form a bilayer structure in order for the membrane to be an insulating barrier. However, a potential ability of the lipids to form nonbilayer structures seems to be a prequisite for several membrane-associated cell processes. Therefore, organisms exposed to changes in the environmental conditions, such as temperature and uptake of fatty acids, adjust their membrane lipid composition. Examples of prokaryotic organisms that have been studied in this respect are the cell wall-less bacterium Acholeplasma laidlawii, the Gram-negative bacterium Escherichia coli, and the Gram-positive bacteria Bacillus megaterium and Clostridium butyricum, and among eukaroytic organisms are found fungi, higher plants, and poikilothermic animals. By synthesizing a proper combination of acyl chain and polar head group structures, the organisms modify the phase transition temperatures of the membrane lipids so that they are maintained in a lamellar liquid crystalline phase, and the formation of a lamellar gel phase as well as reserved nonlamellar phases is avoided. It has been shown that A. laidlawii and E. coli maintain a balance between lamellar-formed and nonlamellar-forming lipids. A growing body of evidence shows that nonlamellar-forming membrane lipids play essential roles in many aspects of membrance functioning. Short-lived nonbilayer structures are probably formed in the processes of fusion and fission of lipid bilayers, and long-lived bilayer structures with a small radius of curvature occur in several types of biological membranes (e.g. smooth endoplasmic reticulum, inner mitochondrial membrane, and prolamellar bodies). The activity of membrane-associated proteins can be modulated by adding detergents or nonlamellar-forming lipids to bilayers. Some examples are the regeneration of denatured bacteriorphodopsin, and the activities of protein kinase C, some phospholipases, and some key lipid synthases involved in the lipid metabolism of eukaryotic cells, A. laidlawii, and E. coli. The physico-chemical properties of the lipid matrix can be a direct feed-back signal on the activity of the lipid synthases. Finally, nonlamellar-forming lipids are essential for a proper cell division, and an efficient translocation of proteins across the plasma membrane, of E. coli cells.

Lipid phase structure in the regulation of lipid composition in Acholeplasma laidlawii membranes.

In Acholeplasma laidlawii membranes the ratio between the dominating lipids, monoglucosyldiglyceride (MGDG) and diglucosyldiglyceride (DGDG), depends on temperature, configuration of incorporated fatty acids, and membrane cholesterol content, which affect the molecular geometry of the lipids. MGDG and DGDG have wedge- and rod-like molecular shapes, respectively, that are modifiable. The packing constraints of lipids in amphiphilic aggregates, i.e., the area of the hydrocarbon-water interface and the volume and length of the hydrocarbon chains, are important in determining the aggregate structure. Pure MGDG forms a reversed hexagonal- (HII) phase structure with different acyl chain contents, while DGDG forms a lamellar phase. Depending on the unsaturated acyl chain content in the lipids, an in vitro mixture of MGDG and DGDG forms lamellar or cubic phases at physiologic temperatures. A high degree of cis-unsaturation, large amounts of MGDG and high temperatures favor formation of the cubic phase. Addition of cholesterol corresponding to the maximal amount incorporable into A. laidlawii induces a transition from a lamellar or a cubic phase to a reversed hexagonal phase. Lipid mixtures containing only unsaturated acyl chains are more sensitive to the bilayer-destabilizing effect of cholesterol than are mixtures with equal amounts of saturated and unsaturated acyl chains. The lamellar phase is the only one compatible with a functional biological membrane. Consequently, the balance between lipids that form lamellar and other mesophase structures must keep within certain limits. The cubic and reversed hexagonal structures were discovered under conditions not existing in the living Acholeplasma cell. Thus, the response of A. laidlawii lipid metabolism to external and internal stimuli can be predicted on the basis of molecular shapes and is necessary to the maintenance of optimal membranes stability. The reduced capacity of Acholeplasma membranes to incorporate cholesterol is a consequence of this regulation.

Lipid bilayer stability in membranes. Regulation of lipid composition in Acholeplasma laidlawii as governed by molecular shape.

The polar lipid composition in membranes of Acholeplasma laidlawii is extensively regulated as a response to environmental changes. In particular, the ratio between the dominating lipids monoglucosyldiglyceride and diglucosyldiglyceride is altered depending on temperature, configuration of incorporated fatty acids, and membrane cholesterol content. Synthesis of monoglucosyldiglyceride is stimulated by low temperature and saturated fatty acids but diminished by the presence of cholesterol. These factors are likely to affect the molecular geometry of the membrane lipids. Monoglucosyldiglyceride and diglucosyldiglyceride have wedge- and rodlike molecular shapes, respectively, that are modifiable to a certain extent. The packing constraints of lipids in amphilphilic aggregates, i.e., hydrocarbon-water interfacial area, hydrocarbon chain volume, and hydrocarbon chain length, are very important in determining the aggregate structure [Israelachvili, J. N. Mitchell, D. J., & Ninham, B. W. (1976) J. Chem. Soc., Faraday Trans. 272, 1525]. Pure monoglucosyldiglyceride forms a reversed hexagonal (HII) phase structure with different fatty acid contents, while diglucosyldiglyceride forms a lamellar phase. However, the only lipid structure compatible with a functional biological membrane is the lamellar phase. Consequently, the balance between lipids forming lamellar and other mesophase structures must keep within certain limits. Here we show that the response in A. liaidlawii lipid metabolism following external and internal stimuli can be predicted on the basis of molecular shapes and is necessary for the cell in order to maintain optimal membrane stability. Furthermore, the reduced capacity of Acholeplasma membranes to incorporate cholesterol is another consequence of this regulation, aiming at preservation of bilayer stability.