Book reviews

The GLUT4 facilitative glucose transporter protein is primarily expressed in muscle and adipose tissue and accounts for the majority of post-prandial glucose uptake. In the basal or non-stimulated state, GLUT4 is localized to intracellular membrane compartments sequestered away from circulating glucose. However, in response to agonist stimulation, there is a marked redistribution of the GLUT4 protein to the cell surface membrane providing a transport route for the uptake of glucose. This GLUT4 translocation can be divided into four general steps: (i) GLUT4 vesicle trafficking outofthe storage pool, (ii) docking just below the cell surface, (iii) priming via the interactions of the SNARE proteins present on the vesicular and plasma membranes, and (iv) fusion of the GLUT4 vesicle with the plasma membrane. This review focuses on recent advances made in identification and characterization of the molecular events and protein interactions involved in these steps of insulin-stimulated GLUT4 translocation.     

Lipid Segregation and Membrane Budding Induced by the Peripheral Membrane Binding Protein Annexin A2

The formation of dynamic membrane microdomains is an important phenomenon in many signal transduction and membrane trafficking events. It is driven by intrinsic properties of membrane lipids and integral as well as membrane-associated proteins. Here we analyzed the ability of one peripherally associated membrane protein, annexin A2 (AnxA2), to induce the formation of phosphatidylinositol 4,5-bisphosphate (PI(4,5)P₂)-rich domains in giant unilamellar vesicles (GUVs) of complex lipid composition. AnxA2 is a cytosolic protein that can bind PI(4,5)P₂ and other acidic phospholipids in a Ca²⁺-dependent manner and that has been implicated in cellular membrane dynamics in endocytosis and exocytosis. We show that AnxA2 binding to GUVs induces lipid phase separation and the recruitment of PI(4,5)P₂, cholesterol and glycosphingolipids into larger clusters. This property is observed for the full-length monomeric protein, a mutant derivative comprising the C-terminal protein core domain and for AnxA2 residing in a heterotetrameric complex with its intracellular binding partner S100A10. All AnxA2 derivatives inducing PI(4,5)P₂ clustering are also capable of forming interconnections between PI(4,5)P₂-rich microdomains of adjacent GUVs. Furthermore, they can induce membrane indentations rich in PI(4,5)P₂ and inward budding of these membrane domains into the lumen of GUVs. This inward vesiculation is specific for AnxA2 and not shared with other PI(4,5)P₂-binding proteins such as the pleckstrin homology (PH) domain of phospholipase Cδ1. Together our results indicate that annexins such as AnxA2 can efficiently induce membrane deformations after lipid segregation, a mechanism possibly underlying annexin functions in membrane trafficking.     

Dynamic modification of sphingomyelin in lipid microdomains controls development of obesity, fatty liver, and type 2 diabetes

Lipid microdomains or caveolae, small invaginations of plasma membrane, have emerged as important elements for lipid uptake and glucose homeostasis. Sphingomyelin (SM) is one of the major phospholipids of the lipid microdomains. In this study, we investigated the physiological function of sphingomyelin synthase 2 (SMS2) using SMS2 knock-out mice, and we found that SMS2 deficiency prevents high fat diet-induced obesity and insulin resistance. Interestingly, in the liver of SMS2 knock-out mice, large and mature lipid droplets were scarcely observed. Treatment with siRNA for SMS2 also decreased the large lipid droplets in HepG2 cells. Additionally, the siRNA of SMS2 decreased the accumulation of triglyceride in liver of leptin-deficient (ob/ob) mice, strongly suggesting that SMS2 is involved in lipid droplet formation. Furthermore, we found that SMS2 exists in lipid microdomains and partially associates with the fatty acid transporter CD36/FAT and with caveolin 1, a scaffolding protein of caveolae. Because CD36/FAT and caveolin 1 exist in lipid microdomains and are coordinately involved in lipid droplet formation, SMS2 is implicated in the modulation of the SM in lipid microdomains, resulting in the regulation of CD36/FAT and caveolae. Here, we established new cell lines, in which we can completely distinguish SMS2 activity from SMS1 activity, and we demonstrated that SMS2 could convert ceramide produced in the outer leaflet of the plasma membrane into SM. Our findings demonstrate the novel and dynamic regulation of lipid microdomains via conformational changes in lipids on the plasma membrane by SMS2, which is responsible for obesity and type 2 diabetes.     

ABC lipid transporters: extruders, flippases, or flopless activators?

Many mammalian ABC transporters move membrane lipids to acceptor lipid assemblies in the extracellular aqueous milieu. Because the desorption from the membrane costs more energy than provided by two ATPs, the transporter probably only translocates the lipid to a partially hydrophilic site on its extracellular face. From this high-energy site, the lipid may efficiently move to the acceptor, which ideally is bound to the transporter, or, in the absence of an acceptor, fall back into the membrane. If the lipid originated from the cytosolic membrane surface, this represents lipid flop and is probably a side activity of the transporters.     

Yeast Lipids Can Phase-separate into Micrometer-scale Membrane Domains

The lipid raft concept proposes that biological membranes have the potential to form functional domains based on a selective interaction between sphingolipids and sterols. These domains seem to be involved in signal transduction and vesicular sorting of proteins and lipids. Although there is biochemical evidence for lipid raft-dependent protein and lipid sorting in the yeast Saccharomyces cerevisiae, direct evidence for an interaction between yeast sphingolipids and the yeast sterol ergosterol, resulting in membrane domain formation, is lacking. Here we show that model membranes formed from yeast total lipid extracts possess an inherent self-organization potential resulting in liquid-disordered-liquid-ordered phase coexistence at physiologically relevant temperature. Analyses of lipid extracts from mutants defective in sphingolipid metabolism as well as reconstitution of purified yeast lipids in model membranes of defined composition suggest that membrane domain formation depends on specific interactions between yeast sphingolipids and ergosterol. Taken together, these results provide a mechanistic explanation for lipid raft-dependent lipid and protein sorting in yeast.     

Gel domains in the plasma membrane of Saccharomyces cerevisiae: highly ordered, ergosterol-free, and sphingolipid-enriched lipid rafts

The plasma membrane of Saccharomyces cerevisiae was studied using the probes trans-parinaric acid and diphenylhexatriene. Diphenylhexatriene anisotropy is a good reporter of global membrane order. The fluorescence lifetimes of trans-parinaric acid are particularly sensitive to the presence and nature of ordered domains, but thus far they have not been measured in yeast cells. A long lifetime typical of the gel phase (>30 ns) was found in wild-type (WT) cells from two different genetic backgrounds, at 24 and 30 °C, providing the first direct evidence for the presence of gel domains in living cells. To understand their nature and location, the study of WT cells was extended to spheroplasts, the isolated plasma membrane, and liposomes from total lipid and plasma membrane lipid extracts (with or without ergosterol extraction by cyclodextrin). It is concluded that the plasma membrane is mostly constituted by ordered domains and that the gel domains found in living cells are predominantly at the plasma membrane and are formed by lipids. To understand their composition, strains with mutations in sphingolipid and ergosterol metabolism and in the glycosylphosphatidylinositol anchor remodeling pathway were also studied. The results strongly indicate that the gel domains are not ergosterol-enriched lipid rafts; they are mainly composed of sphingolipids, possibly inositol phosphorylceramide, and contain glycosylphosphatidylinositol-anchored proteins, suggesting an important role in membrane traffic and signaling, and interactions with the cell wall. The abundance of the sphingolipid-enriched gel domains was inversely related to the cellular membrane system global order, suggesting their involvement in the regulation of membrane properties.     

Sphingolipid Domains in the Plasma Membranes of Fibroblasts Are Not Enriched with Cholesterol

The plasma membranes of mammalian cells are widely expected to contain domains that are enriched with cholesterol and sphingolipids. In this work, we have used high-resolution secondary ion mass spectrometry to directly map the distributions of isotope-labeled cholesterol and sphingolipids in the plasma membranes of intact fibroblast cells. Although acute cholesterol depletion reduced sphingolipid domain abundance, cholesterol was evenly distributed throughout the plasma membrane and was not enriched within the sphingolipid domains. Thus, we rule out favorable cholesterol-sphingolipid interactions as dictating plasma membrane organization in fibroblast cells. Because the sphingolipid domains are disrupted by drugs that depolymerize the cells actin cytoskeleton, cholesterol must instead affect the sphingolipid organization via an indirect mechanism that involves the cytoskeleton.     

Amyloid beta-protein and lipid metabolism

Lipids play an important part as risk or protective factors for Alzheimer's disease. This review summarizes the current findings in which lipids influence Alzheimer's disease and introduces the molecular mechanism how these lipids are linked to amyloid production. Besides the pathological impact of amyloid in Alzheimer's disease, amyloid has a physiological function in regulating lipid homeostasis in return. The understanding of the resulting regulatory cycles between amyloid precursor protein processing and lipids provides a platform for the development of new causal therapeutic approaches for Alzheimer's disease.     

Sphingolipid trafficking and protein sorting in epithelial cells

Sphingolipids represent a minor, but highly dynamic subclass of lipids in all eukaryotic cells. They are involved in functions that range from structural protection to signal transduction and protein sorting, and participate in lipid raft assembly. In polarized epithelial cells, which display an asymmetric apical and basolateral membrane surface, rafts have been proposed as a sorting principle for apical resident proteins, following their biosynthesis. However, raft-mediated trafficking is ubiquitous in cells. Also, sphingolipids per se, which are strongly enriched in the apical domain, are subject to sorting in polarity development. Next to the trans Golgi network, a subapical compartment called SAC or common endosome appears instrumental in regulating these sorting events.     

P4 ATPases - The physiological relevance of lipid flipping transporters

P4 ATPases are integral transmembrane proteins implicated in phospholipid translocation from the exoplasmic to the cytosolic leaflet of biological membranes. Our present knowledge on the cellular physiology of P4 ATPases is mostly derived from studies in the yeast Saccharomyces cerevisiae, where P4 ATPases play a pivotal role in the biogenesis of intracellular transport vesicles, polarized protein transport and protein maturation. In contrast, the physiological and cellular functions of mammalian P4 ATPases are largely unexplored. P4 ATPases act in concert with members of the CDC50 protein family, which are putative β-subunits for P4 ATPases. This review highlights the current status of a slowly emerging research field and emphasizes the contribution of P4 ATPases to the vesicle-generating machinery.     

Detection of lipid domains in model and cell membranes by fluorescence lifetime imaging microscopy

The discovery that the lipids constituting the plasma membrane are not randomly distributed, but instead are able to form laterally segregated lipid domains with different properties has given hints how the formation of such lipid domains influences and regulates many processes occurring at the plasma membrane. While in model systems these lipid domains can be easily accessed and their properties studied, it is still challenging to determine the properties of cholesterol rich lipid domains, the so called “Rafts”, in the plasma membrane of living cells due to their small size and transient nature. One promising technique to address such issues is fluorescence lifetime imaging (FLIM) microscopy, as spatially resolved images make the visualization of the lateral lipid distribution possible, while at the same time the fluorescence lifetime of a membrane probe yields information about the bilayer structure and organization of the lipids in lipid domains and various properties like preferential protein-protein interactions or the enrichment of membrane probes. This review aims to give an overview of the techniques underlying FLIM probes which can be applied to investigate the formation of lipid domains and their respective properties in model membrane and biological systems. Also a short technical introduction into the techniques of a FLIM microscope is given.     

Lipid dependence of ABC transporter localization and function

Lipid rafts have been implicated in many cellular functions, including protein and lipid transport and signal transduction. ATP-binding cassette (ABC) transporters have also been localized in these membrane domains. In this review the evidence for this specific localization will be evaluated and discussed in terms of relevance to ABC transporter function. We will focus on three ABC transporters of the A, B and C subfamily, respectively. Two of these transporters are relevant to multidrug resistance in tumor cells (Pgp/ABCB1 and MRP1/ABCC1), while the third (ABCA1) is extensively studied in relation to the reverse cholesterol pathway and cellular cholesterol homeostasis. We will attempt to derive a generalized model of lipid rafts to which they associate based on the use of various different lipid raft isolation procedures. In the context of lipid rafts, modulation of ABC transporter localization and function by two relevant lipid classes, i.e. sphingolipids and cholesterol, will be discussed.     

Membrane properties of sphingomyelins

Sphingomyelin and phosphatidylcholine are important components in the external leaflet of cellular plasma membranes. In this review we compare the structure of these lipid molecules, with emphasis on the differences in hydrogen bonding capacity and membrane properties that arise from the small but significant differences in molecular structure. The membrane properties of sphingomyelins and the implications that these have, or might have, in biological membranes and for raft function are further discussed.     

Book reviews

Angiogenesis, or the growth of new blood vessels, has, in recent years, become an area of intense scientific research. The primary reason for this has been the realization that angiogenesis plays a key role in many common pathologies, and that its inhibition could have profoound implications in the treatment of these disorders. A substantial number of anti-angiogenic agents have now been identified; however, none has, as of yet, achieved widespread acceptance in the clinic. Many agents have been identified as the result of clearly defined research programs, such as the inhibitors of the vascular endothelial growth factor transmembrane tyrosine kinase receptors, but many other simply by screening. The purpose of this article is to reveiw the wealth of information available on known anti-angiogenic agents and to assess their future potential.     

Book reviews

Hugh Honour, THE IMAGE OF THE BLACK IN WESTERN ART, IV FROM THE AMERICAN REVOLUTION TO WORLD WAR I, Cambridge, Mass. and London: Harvard University Press, 1989; part i, 379 pp., 196 ill., part ii, 306 pp., 183 ill., £34.95 each volume.

Deborah Willis‐Thomas, AN ILLUSTRATED BIO‐BIBLIOGRAPHY OF BLACK PHOTOGRAPHERS, 1940–1988, New York & London: Garland Publishing, Inc., 1989, 483 pp., $85.00.

John Hutchinson, THE DYNAMICS OF CULTURAL NATIONALISM: THE GAELIC REVIVAL AND THE CREATION OF THE IRISH NATION STATE, London: Allen and Unwin, 1987, viii + 343 pp., £38.00.

Arthur Aughey, UNDER SIEGE: ULSTER UNIONISM AND THE ANGLO‐IRISH AGREEMENT, London: Hurst & Co, 1989, xv + 214 pp., £18.50.

Richard Jenkins (ed.), NORTHERN IRELAND: STUDIES IN SOCIAL AND ECONOMIC LIFE, London: Avebury, 1989, xii + 197 pp., £23.50.

J. I. Little, NATIONALISM, CAPITALISM, AND COLONIZATION IN NINETEENTH‐CENTURY QUEBEC: THE UPPER ST. FRANCIS DISTRICT, Kingston, Montreal, London: McGill‐Queen's University Press, 1989, 336 pp., £31.45.

Basdeo Mangru, BENEVOLENT NEUTRALITY: INDIAN GOVERNMENT POLICY AND LABOUR MIGRATION TO BRITISH GUIANA 1854–1884, London: Hansib, 1987, 267 pp., £12.95.

Carole Marks, FAREWELL ‐ WE'RE GOOD AND GONE: THE GREAT BLACK MIGRATION, Bloomington: Indiana University Press, 1989, x + 209 pp., $37.50 and $12.95 (paper).

James R. Grossman, LAND OF HOPE: CHICAGO, BLACK SOUTHERNERS, AND THE GREAT MIGRATION, Chicago: University of Chicago Press, 1989, 384 pp., £23.95.

Robert M. Jiobu, ETHNICITY AND ASSIMILATION: BLACKS, CHINESE, FILIPINOS, JAPANESE, KOREANS, MEXICANS, VIETNAMESE AND WHITES, Albany: State University of New York Press, 1988, np.

John M. MacKenzie (ed.), IMPERIALISM AND POPULAR CULTURE, Manchester: Manchester University Press, 1989, 264 pp., (illus.), £10.95 (paper).

Anne Phillips, THE ENIGMA OF COLONIALISM: BRITISH POLICY IN WEST AFRICA, James Currey and Indiana University Press, vii + 184 pp., £25.00 and £8.95 (paper).

John Darwin, BRITAIN AND DECOLONISATION: THE RETREAT FROM EMPIRE IN THE POST‐WAR WORLD, London: Macmillan, 1988, xiii + 383 pp., £33.00, and £9.95 (paper).

Tony Kushner and Kenneth Lunn (eds), TRADITIONS OF INTOLERANCE: HISTORICAL PERSPECTIVES ON FASCISM AND RACE DISCOURSE IN BRITAIN, Manchester University Press, 1989, 245 pp., £29.95.     

Book reviews

The action of peroxynitrite in vivo has been proposed to account for the involvement of nitrotyrosine in the pathogenesis of many diseases. However, it has been demonstrated that nitrite under acidic conditions, similar to those in the human stomach, also has the ability to nitrate tyrosine. Dietary nitrate is also implicated in the progression of gastritis and gastric cancer and elevated levels of nitrate are found in many disease states in which nitrotyrosine may play a role. Thus, we investigated whether the dietary nitrate intake might contribute towards the plasma protein-bound levels of nitrotyrosine.

Seven healthy, non-smokers participated in a two-day study consisting of a nitrate-low control day followed by a day during which three nitrate-rich meals were consumed. Maximal urinary excretion was attained 4–6 hours after consumption of a meal and the maximum was proportional to the dose. Plasma nitrate was elevated nine-fold, 1 hour after consumption of a meal containing 128.3 mg nitrate. Plasma nitrated protein levels did not appear to alter significantly from basal 1 hour after supplementation with a nitrate-rich meal. Thus dietary nitrate does not appear to contribute to the levels of plasma nitrated proteins, as determined using a competitive inhibition of binding ELISA assay, but this does not preclude any contribution it may make to the total body burden of nitrotyrosine.     

Book reviews

The cichlid fishes of Lake Malawi represent a prime example of a rapid evolutionary radiation via sexual selection. We show that in this group, acoustic communication significantly coincides with visual courtship displays. Specifically, individuals tend to use both multimodal and unimodal displays in successive courtship bouts. Behavioral analyses on six species of Malawian cichlids from two divergent genera revealed that five of the species use displays containing both an auditory and a visual component. Metriaclima zebra “katale” was the notable exception, along with a single individual of Metriaclima lombardoi, which utilized unimodal and multimodal displays with equal frequency. Taken together, these results highlight the importance of behavioral lability in multiple sensory signals at both the intra- and interspecific levels.     

Book reviews

A new medium is described for the growth of the freshwater diatom Melosira italica subsp. subarctica which allows prolonged exponential growth in batch culture. The medium was optimized for cell yield and observations are reported on varying the pH, shaking rate and trace-metal concentrations in the medium. The medium contained no vitamins. Preliminary results suggest that the silica usage was 0·163 mg Si mm^-3 filament volume, the molar ratio of Si:P uptake varied between 46:1 during exponential growth and 30:1 during silica limitation; the molar N:P ratio of the cells averaged 5·6:1.     

Book reviews

A total of 59 taxa of epibionts and endobionts occurred on experimentally deployed gastropod shells within one year of emplacement at depths ranging from 15 m to 260 m in the Bahamas. Most of the diversity occurred within 73 m of water depth. The experimental shells at the deepest sites (210 m, 260 m) were essentially pristine. Differences in experimental treatment affected the results: shells in bags contained more bionts than tethered shells, suggesting the bags had more protective areas for biont settlement. Soft‐bodied encrusters were restricted to the upper 73 m while foraminiferans and bryozoans exhibited bathymetric trends to the deepest sites. While boring algae and cyanobacteria were ubiquitous on the shells to 73 m, other bioeroders (e.g., clionid sponges) were rare. Bioaccumulation, rather than bioerosion, is the predominant process affecting mollusc shells during the first year of taphonomic exposure in carbonate systems to depths of 260 m.