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Showing papers on "Membrane lipids published in 2011"


Journal ArticleDOI
TL;DR: The emerging principles of membrane architecture are reviewed with special emphasis on lipid organization and domain formation, which combines the potential for sphingolipid-cholesterol self-assembly with protein specificity to focus and regulate membrane bioactivity.
Abstract: Cell membranes are composed of a lipid bilayer, containing proteins that span the bilayer and/or interact with the lipids on either side of the two leaflets. Although recent advances in lipid analytics show that membranes in eukaryotic cells contain hundreds of different lipid species, the function of this lipid diversity remains enigmatic. The basic structure of cell membranes is the lipid bilayer, composed of two apposing leaflets, forming a twodimensional liquid with fascinating properties designed to perform the functions cells require. To coordinate these functions, the bilayer has evolved the propensity to segregate its constituents laterally. This capability is based on dynamic liquid‐liquid immiscibility and underlies the raft concept of membrane subcompartmentalization. This principle combines the potential for sphingolipid-cholesterol self-assembly with protein specificity to focus and regulate membrane bioactivity. Here we will review the emerging principles of membrane architecture with special emphasis on lipid organization and domain formation.

931 citations


Reference BookDOI
18 Jul 2011
TL;DR: In this article, the roles of cholesterol in the Biology of cells were discussed and the role of G-protein-protein coupled receptive receptors was discussed. But the authors focused on the structure and function of the G-Protein Coupled Receptors.
Abstract: MEMBRANE LIPID STRUCTURE AND FUNCTION Lipid Structure, Helmut Hauser and Guy Poupart The Mesomorphic Phase Behavior of Lipid Bilayers, Ruthven N.A.H. Lewis, Ronald N. McElhaney Lipid Bilayer Interdigitation, James L. Slater and Ching-hsien Huang The Dynamics of Membrane Lipids, Klaus Gawrisch Non-Lamellar Lipid Phases, Sol M. Gruner The Forces Between Interacting Bilayer Membranes and the Hydration of Phospholipid Assemblies, Richard P. Rand and V.A. Parsegian The Roles of Cholesterol in the Biology of Cells, Philip L. Yeagle Lipid Membrane Fusion, David P. Siegel Membrane Rafts, Kathleen Boesze-Battaglia MEMBRANE PROTEIN STRUCTURE AND FUNCTION Passive and Facilitated Transport, Shinpei Ohki, Robert A. Spangler Membrane Protein Dynamics: Rotational Dynamics, Richard J. Cherry Translational Diffusion of Membrane Proteins, Michael Edidin Inorganic Anion Transporter AE1, Michael L. Jennings The Structures of G-Protein Coupled Receptors, Philip L. Yeagle Role of Membrane Lipids in Modulating the Activity of Membrane-Bound Enzymes, Richard M. Epand Viral Fusion Mechanisms, Aditya Mittal and Joe Bentz

535 citations


Journal ArticleDOI
TL;DR: A shotgun-based lipidomics workflow was developed that enabled the absolute quantification of mammalian membrane lipidomes with minimal sample processing from low sample amounts and investigated the remodeling of the total cell membrane lipidome during the transition from a nonpolarized morphology to an epithelial morphology and vice versa.
Abstract: Tissue differentiation is an important process that involves major cellular membrane remodeling. We used Madin–Darby canine kidney cells as a model for epithelium formation and investigated the remodeling of the total cell membrane lipidome during the transition from a nonpolarized morphology to an epithelial morphology and vice versa. To achieve this, we developed a shotgun-based lipidomics workflow that enabled the absolute quantification of mammalian membrane lipidomes with minimal sample processing from low sample amounts. Epithelial morphogenesis was accompanied by a major shift from sphingomyelin to glycosphingolipid, together with an increase in plasmalogen, phosphatidylethanolamine, and cholesterol content, whereas the opposite changes took place during an epithelial-to-mesenchymal transition. Moreover, during polarization, the sphingolipids became longer, more saturated, and more hydroxylated as required to generate an apical membrane domain that serves as a protective barrier for the epithelial sheet.

426 citations


Journal ArticleDOI
TL;DR: It is now clear that LDs are not an inert store of excess lipids but are dynamically engaged in various cellular functions, some of which are not directly related to lipid metabolism.
Abstract: Lipid droplets (LDs) are independent organelles that are composed of a lipid ester core and a surface phospholipid monolayer. Recent studies have revealed many new proteins, functions, and phenomena associated with LDs. In addition, a number of diseases related to LDs are beginning to be understood at the molecular level. It is now clear that LDs are not an inert store of excess lipids but are dynamically engaged in various cellular functions, some of which are not directly related to lipid metabolism. Compared to conventional membrane organelles, there are still many uncertainties concerning the molecular architecture of LDs and how each function is placed in a structural context. Recent findings and remaining questions are discussed.

404 citations


Journal ArticleDOI
TL;DR: This study reports that, independent of inflammasome and membrane proteins, alum binds dendritic cell (DC) plasma membrane lipids with substantial force and proposes that alum triggers DC responses by altering membrane lipid structures.
Abstract: As an approved vaccine adjuvant for use in humans, alum has vast health implications, but, as it is a crystal, questions remain regarding its mechanism. Furthermore, little is known about the target cells, receptors, and signaling pathways engaged by alum. Here we report that, independent of inflammasome and membrane proteins, alum binds dendritic cell (DC) plasma membrane lipids with substantial force. Subsequent lipid sorting activates an abortive phagocytic response that leads to antigen uptake. Such activated DCs, without further association with alum, show high affinity and stable binding with CD4(+) T cells via the adhesion molecules intercellular adhesion molecule-1 (ICAM-1) and lymphocyte function-associated antigen-1 (LFA-1). We propose that alum triggers DC responses by altering membrane lipid structures. This study therefore suggests an unexpected mechanism for how this crystalline structure interacts with the immune system and how the DC plasma membrane may behave as a general sensor for solid structures.

364 citations


Journal ArticleDOI
TL;DR: Upon both acid and base hydrolyses of total cell material, the uncommon membrane-spanning lipid 13,16-dimethyl octacosanedioic acid (iso-diabolic acid) was released in substantial amounts (22 to 43% of the total fatty acids) from all of the acidobacteria studied.
Abstract: The distribution of membrane lipids of 17 different strains representing 13 species of subdivisions 1 and 3 of the phylum Acidobacteria, a highly diverse phylum of the Bacteria, were examined by hydrolysis and gas chromatography-mass spectrometry (MS) and by high-performance liquid chromatography-MS of intact polar lipids. Upon both acid and base hydrolyses of total cell material, the uncommon membrane-spanning lipid 13,16-dimethyl octacosanedioic acid (iso-diabolic acid) was released in substantial amounts (22 to 43% of the total fatty acids) from all of the acidobacteria studied. This lipid has previously been encountered only in thermophilic Thermoanaerobacter species but bears a structural resemblance to the alkyl chains of bacterial glycerol dialkyl glycerol tetraethers (GDGTs) that occur ubiquitously in peat and soil and are suspected to be produced by acidobacteria. As reported previously, most species also contained iso-C15 and C16:1ω7C as major fatty acids but the presence of iso-diabolic acid was unnoticed in previous studies, most probably because the complex lipid that contained this moiety was not extractable from the cells; it could only be released by hydrolysis. Direct analysis of intact polar lipids in the Bligh-Dyer extract of three acidobacterial strains, indeed, did not reveal any membrane-spanning lipids containing iso-diabolic acid. In 3 of the 17 strains, ether-bound iso-diabolic acid was detected after hydrolysis of the cells, including one branched GDGT containing iso-diabolic acid-derived alkyl chains. Since the GDGT distribution in soils is much more complex, branched GDGTs in soil likely also originate from other (acido)bacteria capable of biosynthesizing these components.

360 citations


Journal ArticleDOI
TL;DR: A rapid and efficient system for the production of large quantities of affinity-purified HCV allowing a comprehensive analysis of the infectious virion, including the determination of its lipid composition is described.

348 citations


Journal ArticleDOI
TL;DR: This study provides the first biochemical evidence for the involvement of an essential protein in the transport of lipid‐linked cell wall precursors across biogenic membranes.
Abstract: Bacterial cell growth necessitates synthesis of peptidoglycan. Assembly of this major constituent of the bacterial cell wall is a multistep process starting in the cytoplasm and ending in the exterior cell surface. The intracellular part of the pathway results in the production of the membrane-anchored cell wall precursor, Lipid II. After synthesis this lipid intermediate is translocated across the cell membrane. The translocation (flipping) step of Lipid II was demonstrated to require a specific protein (flippase). Here, we show that the integral membrane protein FtsW, an essential protein of the bacterial division machinery, is a transporter of the lipid-linked peptidoglycan precursors across the cytoplasmic membrane. Using Escherichia coli membrane vesicles we found that transport of Lipid II requires the presence of FtsW, and purified FtsW induced the transbilayer movement of Lipid II in model membranes. This study provides the first biochemical evidence for the involvement of an essential protein in the transport of lipid-linked cell wall precursors across biogenic membranes.

329 citations


Journal ArticleDOI
TL;DR: The importance of the lipid composition of bacterial membranes in determining the susceptibility of the organism to the action of certain antimicrobial agents is emphasized.
Abstract: Many antimicrobial agents that target bacteria are cationic and can interact with the anionic lipid components that are exposed on the bacterial membrane. Bacteria vary widely in the nature of the major lipid components that are in the cell membrane. Those bacteria with both anionic as well as zwitterionic or neutral lipids can be induced to form domains in the presence of antimicrobial peptides possessing several cationic charges. This segregation of anionic and zwitterionic lipids into domains can result in the arrest of cell growth or in cell death. Such agents are generally more toxic to Gram-negative bacteria, than to Gram-positive ones. These findings emphasize the importance of the lipid composition of bacterial membranes in determining the susceptibility of the organism to the action of certain antimicrobial agents.

270 citations


Journal ArticleDOI
21 Oct 2011-Science
TL;DR: It is shown that rotary adenosine triphosphatases (ATPases)/synthases from Thermus thermophilus and Enterococcus hirae can be maintained intact with membrane and soluble subunit interactions preserved in vacuum and can link specific lipid and nucleotide binding with distinct regulatory roles.
Abstract: The ability of electrospray to propel large viruses into a mass spectrometer is established and is rationalized by analogy to the atmospheric transmission of the common cold. Much less clear is the fate of membrane-embedded molecular machines in the gas phase. Here we show that rotary adenosine triphosphatases (ATPases)/synthases from Thermus thermophilus and Enterococcus hirae can be maintained intact with membrane and soluble subunit interactions preserved in vacuum. Mass spectra reveal subunit stoichiometries and the identity of tightly bound lipids within the membrane rotors. Moreover, subcomplexes formed in solution and gas phases reveal the regulatory effects of nucleotide binding on both ATP hydrolysis and proton translocation. Consequently, we can link specific lipid and nucleotide binding with distinct regulatory roles.

249 citations


Journal ArticleDOI
TL;DR: The PP-LFERs revealed that partition coefficients into membrane lipids can be two log units higher than those into storage lipids for H-bond donor compounds, suggesting that distinguishing between the two lipids is necessary to account for the bioaccumulation of these compounds.
Abstract: Lipids have been considered as the predominant components for bioaccumulation of organic chemicals. However, differences in accumulation properties between different types of lipid (e.g., storage and membrane lipids) have rarely been considered. Moreover, in view of toxic effects on organisms, chemical accumulation specifically in biological membranes is of particular importance. In this review article, partition coefficients of 240 neutral organic compounds between liposomes (phospholipid membrane vesicles) and water (Klipw), reported in the literature or measured additionally for this work, were evaluated. Values of log Klipw and log Kow (octanol–water partition coefficients) differ by 0.4 on average. Polyparameter linear free energy relationships (PP-LFERs) can describe the log Klipw data even better (standard deviations = 0.28–0.31) than the log Kow model. Recent experimental data for highly hydrophobic compounds fit well to the PP-LFERs and do not indicate the existence of a previously postulated “hy...

Journal ArticleDOI
TL;DR: Computer simulations and optical microscopy are used to study the sorting of transmembrane helices into the liquid-disordered domains of phase-separated model membranes, irrespective of peptide–lipid hydrophobic mismatch.
Abstract: Cell membranes are comprised of multicomponent lipid and protein mixtures that exhibit a complex partitioning behavior. Regions of structural and compositional heterogeneity play a major role in the sorting and self-assembly of proteins, and their clustering into higher-order oligomers. Here, we use computer simulations and optical microscopy to study the sorting of transmembrane helices into the liquid-disordered domains of phase-separated model membranes, irrespective of peptide-lipid hydrophobic mismatch. Free energy calculations show that the enthalpic contribution due to the packing of the lipids drives the lateral sorting of the helices. Hydrophobic mismatch regulates the clustering into either small dynamic or large static aggregates. These results reveal important molecular driving forces for the lateral organization and self-assembly of transmembrane helices in heterogeneous model membranes, with implications for the formation of functional protein complexes in real cells.

Journal ArticleDOI
TL;DR: It is demonstrated that surprisingly low levels of phospholipids can drive protocell membrane growth during competition for single-chain lipids, and could have been a deterministic outcome of intrinsic physical processes and a key driving force for early cellular evolution.
Abstract: To understand the emergence of Darwinian evolution, it is necessary to identify physical mechanisms that enabled primitive cells to compete with one another. Whereas all modern cell membranes are composed primarily of diacyl or dialkyl glycerol phospholipids, the first cell membranes are thought to have self-assembled from simple, single-chain lipids synthesized in the environment. We asked what selective advantage could have driven the transition from primitive to modern membranes, especially during early stages characterized by low levels of membrane phospholipid. Here we demonstrate that surprisingly low levels of phospholipids can drive protocell membrane growth during competition for single-chain lipids. Growth results from the decreasing fatty acid efflux from membranes with increasing phospholipid content. The ability to synthesize phospholipids from single-chain substrates would have therefore been highly advantageous for early cells competing for a limited supply of lipids. We show that the resulting increase in membrane phospholipid content would have led to a cascade of new selective pressures for the evolution of metabolic and transport machinery to overcome the reduced membrane permeability of diacyl lipid membranes. The evolution of phospholipid membranes could thus have been a deterministic outcome of intrinsic physical processes and a key driving force for early cellular evolution.

Journal ArticleDOI
TL;DR: Viruses intricately interact with and modulate cellular membranes at several stages of their replication, but much less is known about the role of viral lipids compared to proteins and nucleic acids.
Abstract: Viruses intricately interact with and modulate cellular membranes at several stages of their replication, but much less is known about the role of viral lipids compared to proteins and nucleic acids All animal viruses have to cross membranes for cell entry and exit, which occurs by membrane fusion (in enveloped viruses), by transient local disruption of membrane integrity, or by cell lysis Furthermore, many viruses interact with cellular membrane compartments during their replication and often induce cytoplasmic membrane structures, in which genome replication and assembly occurs Recent studies revealed details of membrane interaction, membrane bending, fission, and fusion for a number of viruses and unraveled the lipid composition of raft-dependent and -independent viruses Alterations of membrane lipid composition can block viral release and entry, and certain lipids act as fusion inhibitors, suggesting a potential as antiviral drugs Here, we review viral interactions with cellular membranes important for virus entry, cytoplasmic genome replication, and virus egress

Journal ArticleDOI
TL;DR: It is shown that Arabidopsis has two classes of ceramide synthases discriminating acyl chain length and also that very-long-acyl-chain sphingolipids are required for polar auxin transport in particular during lateral root emergence and a trafficking pathway with specific endomembrane compartments and polar auxIn transport protein cargoes is defined.
Abstract: Sphingolipids are a class of structural membrane lipids involved in membrane trafficking and cell polarity. Functional analysis of the ceramide synthase family in Arabidopsis thaliana demonstrates the existence of two activities selective for the length of the acyl chains. Very-long-acyl-chain (C > 18 carbons) but not long-chain sphingolipids are essential for plant development. Reduction of very-long-chain fatty acid sphingolipid levels leads in particular to auxin-dependent inhibition of lateral root emergence that is associated with selective aggregation of the plasma membrane auxin carriers AUX1 and PIN1 in the cytosol. Defective targeting of polar auxin carriers is characterized by specific aggregation of Rab-A2a– and Rab-A1e–labeled early endosomes along the secretory pathway. These aggregates correlate with the accumulation of membrane structures and vesicle fragmentation in the cytosol. In conclusion, sphingolipids with very long acyl chains define a trafficking pathway with specific endomembrane compartments and polar auxin transport protein cargoes.

Journal ArticleDOI
TL;DR: This method for creating giant vesicles can be used to test models of biological processes that depend on confined volume and complex membrane composition, and it may be useful in constructing functional systems for therapeutic and biomaterials applications.
Abstract: Growing knowledge of the key molecular components involved in biological processes such as endocytosis, exocytosis, and motility has enabled direct testing of proposed mechanistic models by reconstitution. However, current techniques for building increasingly complex cellular structures and functions from purified components are limited in their ability to create conditions that emulate the physical and biochemical constraints of real cells. Here we present an integrated method for forming giant unilamellar vesicles with simultaneous control over (i) lipid composition and asymmetry, (ii) oriented membrane protein incorporation, and (iii) internal contents. As an application of this method, we constructed a synthetic system in which membrane proteins were delivered to the outside of giant vesicles, mimicking aspects of exocytosis. Using confocal fluorescence microscopy, we visualized small encapsulated vesicles docking and mixing membrane components with the giant vesicle membrane, resulting in exposure of previously encapsulated membrane proteins to the external environment. This method for creating giant vesicles can be used to test models of biological processes that depend on confined volume and complex membrane composition, and it may be useful in constructing functional systems for therapeutic and biomaterials applications.

Journal ArticleDOI
TL;DR: This Perspective focuses on the interface of integral transmembrane proteins and membrane lipids in eukaryotic cells and how proteins and lipids interact with each other will be important for unraveling membrane protein structure and function.

Journal ArticleDOI
TL;DR: Two membrane curvature–sensing molecules with opposite chemistries are targeted to distinct vesicle classes through direct interaction with different lipid environments.
Abstract: Membrane curvature sensors have diverse structures and chemistries, suggesting that they might have the intrinsic capacity to discriminate between different types of vesicles in cells. In this paper, we compare the in vitro and in vivo membrane-binding properties of two curvature sensors that form very different amphipathic helices: the amphipathic lipid-packing sensor (ALPS) motif of a Golgi vesicle tether and the synaptic vesicle protein α-synuclein, a causative agent of Parkinson's disease. We demonstrate the mechanism by which α-synuclein senses membrane curvature. Unlike ALPS motifs, α-synuclein has a poorly developed hydrophobic face, and this feature explains its dual sensitivity to negatively charged lipids and to membrane curvature. When expressed in yeast cells, these two curvature sensors were targeted to different classes of vesicles, those of the early secretory pathway for ALPS motifs and to negatively charged endocytic/post-Golgi vesicles in the case of α-synuclein. Through structures with complementary chemistries, α-synuclein and ALPS motifs target distinct vesicles in cells by direct interaction with different lipid environments.

Journal ArticleDOI
TL;DR: It is argued that the packing preferences of the membrane-spanning α-helices in a membrane protein result in a structure that matches nicely with that of the surrounding lipid bilayer, so that lipid and protein can meet without either having to change very much.
Abstract: Intrinsic membrane proteins are solvated by a shell of lipid molecules interacting with the membrane-penetrating surface of the protein; these lipid molecules are referred to as annular lipids. Lipid molecules are also found bound between transmembrane α-helices; these are referred to as non-annular lipids. Annular lipid binding constants depend on fatty acyl chain length, but the dependence is less than expected from models based on distortion of the lipid bilayer alone. This suggests that hydrophobic matching between a membrane protein and the surrounding lipid bilayer involves some distortion of the transmembrane α-helical bundle found in most membrane proteins, explaining the importance of bilayer thickness for membrane protein function. Annular lipid binding constants also depend on the structure of the polar headgroup region of the lipid, and hotspots for binding anionic lipids have been detected on some membrane proteins; binding of anionic lipid molecules to these hotspots can be functionally important. Binding of anionic lipids to non-annular sites on membrane proteins such as the potassium channel KcsA can also be important for function. It is argued that the packing preferences of the membrane-spanning α-helices in a membrane protein result in a structure that matches nicely with that of the surrounding lipid bilayer, so that lipid and protein can meet without either having to change very much.

Journal ArticleDOI
TL;DR: Results implicate alterations in lipid,,,&olism in the development of physical dependence on ethanol in mice kept under identical conditions except that ethanol was not present in the inspired air.
Abstract: ontinuous exposure of mice to increasing conozztions of ethanol vapour is associated first with the lopment of tolerance to ethanol and, after more nged administration, with physical dependence on 4anol (Griffiths, Littleton & Ortiz, 1974). The changes the central nervous system which lead to tolerance zd dependence are unknown, but Chin & Goldstein (1977) have reported that tolerance to the physical cBcctS .of ethanol can be demonstrated in the isolated flaPt,c membranes of ethanol-dependent mice. (1976) has suggested that development of ,lerance to ethanol by bacteria follows the incorporation of fatty acids of different degrees of unsaturation cell membrane phospholipids. Our own results (A(,,, Murad, Begg & others, 1977; Griffiths, Abu ,,gurad & Littleton, 1977) implicate alterations in lipid ,,,&olism in the development of physical dependence on ethanol. It seemed important to establish whether d m g e s in the fatty acid composition of synaptosomal membranes of mouse brain occurred a t any stage during the continuous administration of ethanol. Mice of the T O Swiss strain (LAB, Dagenham, asex) were exposed to ethanol vapour in the way p~viously described (Griffiths & others, 1974). Mice *re killed (plunging into liquid nitrogen) either 2 h rrfter commencing inhalation of ethanol, when no signs of physical dependence are seen, or 10 days after commencing inhalation, when withdrawal of ethanol results in a physical withdrawal syndrome. Brains from these ethanol-treated animals were compared with those of mice kept under identical conditions except that ethanol was not present in the inspired air. Ethanol, in the concentration obtained in brains of treated mice, was added to some control brains to establish the effect of the presence of ethanol on the measurement of fatty acids. Individual whole mouse brains were homogenized in ioGcold 0.32 M sucrose and centrifuged a t SOOg for 10 min. The supernatant was recentrifuged a t 25 OOOg for 30 min at 0\" and the crude synaptosomal fraction in the Pellet subjected to Folch extraction. Phospholipids were adsorbed onto a silicic acid slurry and then eluted Mth methanol. After alkaline hydrolysis the fatty acids methylated by boiling for 2 min with boron tlfluoride-methanol reagent (BDH). After extraction W h chloroform and washing with distilled water, the -thYlesters were dried under nitrogen and stored in the &kat4\" (maximum period 4 h) before chromatography. acid methylesters were separated by gas-liquid cbomatography (N, as carrier a t 70 ml min-' through a prolo

Journal ArticleDOI
TL;DR: The physiological function of sphingomyelin synthase 2 (SMS2) is investigated using SMS2 knock-out mice, and it is found that SMS2 deficiency prevents high fat diet-induced obesity and insulin resistance, and SMS2 could convert ceramide produced in the outer leaflet of the plasma membrane into SM.

Journal ArticleDOI
TL;DR: The steady state asymmetry of the lipids can be disrupted within seconds by the activation of phospholipases and scramblases and the active translocation also contributes to the generation of curvature in the budding of transport vesicles.
Abstract: Cells have thousands of different lipids. In the plasma membrane, and in membranes of the late secretory and endocytotic pathways, these lipids are not evenly distributed over the two leaflets of the lipid bilayer. The basis for this transmembrane lipid asymmetry lies in the fact that glycerolipids are primarily synthesized on the cytosolic and sphingolipids on the noncytosolic surface of cellular membranes, that cholesterol has a higher affinity for sphingolipids than for glycerolipids. In addition, P4-ATPases, “flippases,” actively translocate the aminophospholipids phosphatidylserine and phosphatidylethanolamine to the cytosolic surface. ABC transporters translocate lipids in the opposite direction but they generally act as exporters rather than “floppases.” The steady state asymmetry of the lipids can be disrupted within seconds by the activation of phospholipases and scramblases. The asymmetric lipid distribution has multiple implications for physiological events at the membrane surface. Moreover, the active translocation also contributes to the generation of curvature in the budding of transport vesicles.

Journal ArticleDOI
TL;DR: How these new bioorthogonal ligation reactions and lipid chemical reporters afford new opportunities for exploring the biology of lipid-modified proteins and lipid trafficking are discussed.
Abstract: Protein lipidation and lipid trafficking control many key biological functions in all kingdoms of life. The discovery of diverse lipid species and their covalent attachment to many proteins has revealed a complex and regulated network of membranes and lipidated proteins that are central to fundamental aspects of physiology and human disease. Given the complexity of lipid trafficking and the protein targeting mechanisms involved with membrane lipids, precise and sensitive methods are needed to monitor and identify these hydrophobic molecules in bacteria, yeast, and higher eukaryotes. Although many analytical methods have been developed for characterizing membrane lipids and covalently modified proteins, traditional reagents and approaches have limited sensitivity, do not faithfully report on the lipids of interest, or are not readily accessible. The invention of bioorthogonal ligation reactions, such as the Staudinger ligation and azide-alkyne cycloadditions, has provided new tools to address these limitations, and their use has begun to yield fresh insight into the biology of protein lipidation and lipid trafficking. In this Account, we discuss how these new bioorthogonal ligation reactions and lipid chemical reporters afford new opportunities for exploring the biology of lipid-modified proteins and lipid trafficking. Lipid chemical reporters from our laboratory and several other research groups have enabled improved detection and large-scale proteomic analysis of fatty-acylated and prenylated proteins. For example, fatty acid and isoprenoid chemical reporters in conjunction with bioorthogonal ligation methods have circumvented the limited sensitivity and hazards of radioactive analogues, allowing rapid and robust fluorescent detection of lipidated proteins in all organisms tested. These chemical tools have revealed alterations in protein lipidation in different cellular states and are beginning to provide unique insights in mechanisms of regulation. Notably, the purification of proteins labeled with lipid chemical reporters has allowed both the large-scale analysis of lipidated proteins as well as the discovery of new lipidated proteins involved in metabolism, gene expression, and innate immunity. Specific lipid reporters have also been developed to monitor the trafficking of soluble lipids; these species are enabling bioorthogonal imaging of membranes in cells and tissues. Future advances in bioorthogonal chemistry, specific lipid reporters, and spectroscopy should provide important new insight into the functional roles of lipidated proteins and membranes in biology.

Journal ArticleDOI
TL;DR: It is shown in various mammalian cell lines that both enzymes additionally localize to lipid droplets, which consist of a core of neutral lipids surrounded by a monolayer of phospholipid, mainly PC, and that LDs have the ability to locally synthesize PC.

Journal ArticleDOI
TL;DR: Two types of model for the adaptation of plants to temperature change are proposed: (1) remodelling of membrane lipids but maintenance of the degree of unsaturation are used to adapt to frequent temperature alterations; and (2) both remodelling and changes in the degreeof unsaturation to Adapt to infrequent temperature alterations.
Abstract: One major strategy by which plants adapt to temperature change is to decrease the degree of unsaturation of membrane lipids under high temperature and increase it under low temperature. We hypothesize that this strategy cannot be adopted by plants in ecosystems and environments with frequent alterations between high and low temperatures, because changes in lipid unsaturation are complex and require large energy inputs. To test this hypothesis, we used a lipidomics approach to profile changes in molecular species of membrane glycerolipids in two plant species sampled from alpine screes and in another two plant species grown in a growth chamber, with the temperature cycling daily between heat and freezing. We found that six classes of phospholipid and two classes of galactolipid showed significant changes, but the degree of unsaturation of total lipids and of three lysophospholipid classes remained unchanged. This pattern of changes in membrane lipids was distinct from that occurring during slow alterations in temperature. We propose two types of model for the adaptation of plants to temperature change: (1) remodelling of membrane lipids but maintenance of the degree of unsaturation are used to adapt to frequent temperature alterations; and (2) both remodelling and changes in the degree of unsaturation to adapt to infrequent temperature alterations.

Journal ArticleDOI
TL;DR: Measurements of capsaicin-activated currents in excised patches from TRPV1-expressing HEK cells show that enrichment with cholesterol, but not its diastereoisomer epicholesterol, markedly decreased wild-type rat TRpV1 currents, suggesting that there is a cholesterol-binding site in TRPv1 and that the functions of TRP V1 depend on the genetic variant and membrane cholesterol content.

Journal ArticleDOI
TL;DR: In vitro experiments are presented that highlight the essential physics of lipid sorting at thermal equilibrium: It is driven by a trade-off between bending energy, mixing entropy, and interactions between species.
Abstract: It has often been suggested that the high curvature of transport intermediates in cells may be a sufficient means to segregate different lipid populations based on the relative energy costs of forming bent membranes. In this review, we present in vitro experiments that highlight the essential physics of lipid sorting at thermal equilibrium: It is driven by a trade-off between bending energy, mixing entropy, and interactions between species. We collect evidence that lipid sorting depends strongly on lipid–lipid and protein–lipid interactions, and hence on the underlying composition of the membrane and on the presence of bound proteins.

Journal ArticleDOI
TL;DR: It is suggested that nitration of Y125/133/136 reduces the membrane-binding affinity of aS through allosteric coupling by altering the ensemble of conformational states and depopulating those capable of membrane binding.
Abstract: Both oxidative stress and aggregation of the protein α-synuclein (aS) have been implicated as key factors in the etiology of Parkinson’s disease. Specifically, oxidative modifications to aS disrupt its binding to lipid membranes, an interaction considered critical to its native function. Here we seek to provide a mechanistic explanation for this phenomenon by investigating the effects of oxidative nitration of tyrosine residues on the structure of aS and its interaction with lipid membranes. Membrane binding is mediated by the first ∼95 residues of aS. We find that nitration of the single tyrosine (Y39) in this domain disrupts binding due to electrostatic repulsion. Moreover, we observe that nitration of the three tyrosines (Y125/133/136) in the C-terminal domain is equally effective in perturbing binding, an intriguing result given that the C-terminus is not thought to interact directly with the lipid bilayer. Our investigations show that tyrosine nitration results in a change of the conformational state...

Journal ArticleDOI
TL;DR: In low-phosphorus (P) marine systems, phytoplankton replace membrane phospholipids with nonphosphorous lipids, but it is not known how rapidly this substitution occurs as mentioned in this paper.
Abstract: In low-phosphorus (P) marine systems, phytoplankton replace membrane phospholipids with non-phosphorus lipids, but it is not known how rapidly this substitution occurs. Here, when cells of the model diatom Thalassiosira pseudonana were transferred from P-replete medium to P-free medium, the phospholipid content of the cells rapidly declined within 48 h from 45±0.9 to 21±4.5% of the total membrane lipids; the difference was made up by non-phosphorus lipids. Conversely, when P-limited T. pseudonana were resupplied with P, cells reduced the percentage of their total membrane lipids contributed by a non-phosphorus lipid from 43±1.5 to 7.3±0.9% within 24 h, whereas the contribution by phospholipids rose from 2.2±0.1 to 44±3%. This dynamic phospholipid reservoir contained sufficient P to synthesize multiple haploid genomes, suggesting that phospholipid turnover could be an important P source for cells. Field observations of phytoplankton lipid content may thus reflect short-term changes in P supply and cellular physiology, rather than simply long-term adjustment to the environment.

Journal ArticleDOI
TL;DR: It is revealed that both the hydrophobic and hydrophilic faces of the helix greatly influence binding and sensing, suggesting that the description of membrane curvature-sensing requires consideration of several factors such as short and long range electrostatic interactions, hydrogen bonding, and the volume and structure of insertedHydrophobic residues.