Rapid degradation and limited synthesis of phospholipids in the cotyledons of mung bean seedlings

Seedling growth of mung bean is accompanied by the rapid catabolism of the three major phospholipids in the cotyledons (phosphatidylcholine, phosphatidylethanolamine, and phosphatidylinositol). The decline starts 24 hours after the beginning of imbibition and by the 4th day of growth more than 50% of the phospholipids have been catabolized. Extracts of cotyledons of 24-hour-imbibed beans contain enzymes capable of degrading membrane-associated phospholipids in vitro. This degradation involves phospholipase D and phosphatase activity.     

Susceptibility of plasmenyl glycerophosphoethanolamine lipids containing arachidonate to oxidative degradation

Plasmenyl phospholipids (1-alk-1′-enyl-2-acyl-3-glycerophospholipids, plasmalogens) are a structurally unique class of lipids that contain an α-unsaturated ether substituent at the sn-1 position of the glycerol backbone. Several studies have supported the hypothesis that plasmalogens may be antioxidant molecules that protect cells from oxidative stress. Because the molecular mechanisms responsible for the antioxidant properties of plasmenyl phospholipids are not fully understood, the oxidation of plasmalogens in natural mixtures of phospholipids was studied using electrospray tandem mass spectrometry. Glycerophosphoethanolamine (GPE) lipids from bovine brain were found to contain six major molecular species (16:0p/18:1-, 18:1p/18:1-, 18:0p/20:4-, 16:0p/20:4, 18:0a/20:4-, and 18:0a/22:6-GPE). Oxidation of GPE yielded lyso phospholipid products derived from plasmalogen species containing only monounsaturated sn-2 substituents and diacyl-GPE with oxidized polyunsaturated fatty acyl substituents at sn-2. The only plasmalogen species remaining intact following oxidation contained monounsaturated fatty acyl groups esterified at sn-2. The mechanism responsible for the rapid and specific destruction of plasmalogen GPE may likely involve unique reactivity imparted by a polyunsaturated fatty acyl group esterified at sn-2. This structural feature may play a central role determining the antioxidant properties ascribed to this class of phospholipids.     

Lipid analysis of cells and chromatophores of Rhodopseudomonas sphaeroides

The phospholipids and fatty acid analysis of four strains of Rhodopseudomonas sphaeroides and of chromatophores from two strains show some differences and also show the presence of an unusual polar neutral lipid which is ninhydrin positive and which on acid hydrolysis yields ornithine and an unidentified amino compound. This lipid is called aminolipid-X and has a fatty acid composition very different from the phospholipids. Phosphatidylcholine (PC) and phosphatidylethanolamine (PE) contain a very small amount of plasmalogen forms as determined by combined mild alkaline hydrolysis, acetic acid hydrolysis and phospholipase A₂ hydrolysis.The reaction of intact cells and chromatophores with trinitrobenzenesulfonate (TNBS), fluorodinitrobenzene (FDNB) and isethionylacetimidate (IA) show that 78% of the total PE in chromatophores is localized on the outer membrane surface. In intact cells about 15–35% of the total PE is localized on the outer surface of the plasma membrane.     

Etude comparee des lipides et de la fixation passive du calcium dans les racines et les fractions subcellulaires du Lupinus luteus et de la Vicia faba

In both lupin and broad bean, the root lipids contain paraffins, triglycerides, diglycerides, free fatty acids and polar lipids (phospholipids and galactolipids). The polar lipids and the triglycerides are the more abundant classes. The root galactolipids are mono- and di-galactosyldiglycerides; two steryl glycosides are also present. The phospholipids in both species are: phosphatidylinositol, phosphatidylcholine, phosphatidylglycerol, phosphatidylserine, phosphatidylethanolamine, diphosphatidylglycerol and phosphatidic acid. This last phospholipid represents 8·3% of total lipid phosphorus in Lupinus against 2·3% in Vicia. The other acidic phospholipids represent 30·4% in Lupinus against 20·9% in Vicia. The lipids of Lupinus are rich in linolenic acid whereas those found in Vicia are richer in linoleic acid. The various subcellular fractions prepared from the roots of both species have an homogeneous lipid composition, reflecting exactly that of entire cells. The calcium passive fixation capacity in microsomes and mitochondria of Lupinus roots is more important than that in the same organelles of Vicia faba roots. Thus a relationship is suggested between the amount of phospholipids in membranes and the passive fixation of calcium.     

Activation of a specific phospholipid fatty acyl hydrolase in Dunaliella salina microsomes during acclimation to low growth temperature

Dunaliella salina microsomes, but not chloroplasts, contain a fatty acyl hydrolase with high activity towards endogenous or exogenous phosphatidylglycerol and phosphatidylethanolamine. There is relatively little activity towards other phospholipids or added monogalactosyldiacylglycerol. Lipolysis is most active in the presence of 10 mM Ca²⁺ and is enhanced by added calmodulin. Microsomal acyl hydrolase activity in 30°C-grown cells is low when measured in vitro at 12°C but increases rapidly after cells are chilled to 12°C, finally attaining more than 13-times the pre-chilling value. The changing capacity for lipolysis may explain the key role of microsomes in the acclimation of Dunaliella to low temperature.     

Construction of a lethal mutation in the synthesis of the major acidic phospholipids of Escherichia coli

In order to determine if the major acidic phospholipids of Escherichia coli are essential to the organism, we constructed a null allele (pgsA30) of the pgsA gene thus rendering the organism incapable of synthesizing phosphatidylglycerol or cardiolipin. In strains carrying the pgsA30 allele cell viability, synthesis of gene product and the ability to synthesize the two major acidic phospholipids were dependent on the presence of a functional copy of the pgsA gene carried on a plasmid which was temperature-sensitive for replication. Growth ceased at the temperature restrictive for plasmid replication when the acidic phospholipid content dropped to about 10% of wild type levels which is slightly higher than the level reported in cells carrying the pgsA3 allele in a genetic background derived from strain SD12; the latter cells, which are capable of synthesizing low levels of acidic phospholipids, were previously shown to have no abnormal growth phenotype (Miyazaki, C., Kuroda, M., Ohta, A., and Shibuya, I. (1985) Proc. Natl. Acad. Sci. U. S. A. 82, 7530-7534). The pgsA30 allele, unlike the pgsA3 allele, could not support growth in strain SD12. Neither allele could support growth in two other independently derived strains of E. coli. Therefore, there is a direct dependence of cell viability on a functional pgsA gene product. Strain SD12 appears to contain a suppressor which allows cells with a reduced capability to synthesize acidic phospholipid (pgsA3 allele) to grow, but cannot support growth in cells with a complete lack of synthetic capability (pgsA30 allele).     

Phospholipid and cholesterol uptake by mycoplasma cells and membranes

The ability of growing mycoplasma cells and their isolated membranes to take up exogenous phospholipids was correlated with their ability to take up cholesterol. Horse serum or vesicles made of phosphatidylcholine and cholesterol served as lipid donors. Growing cells of five Mycoplasma species took up significant quantities of phosphatidylcholine and sphingomyelin as well as free and esterified cholesterol. In contrast, growing cells of three Acholeplasma species failed to take up any of the exogenous phospholipids, and only incorporated low amounts of free cholesterol and no esterified cholesterol. Hence, the ability of mycoplasmas to take up large quantities of cholesterol appears to be correlated with an ability to take up exogenous phospholipids. Isolated membranes of Mycoplasma capricolum and Acholeplasma laidlawii took up lower amounts of cholesterol than did membranes of growing cells and did not take up phospholipids. Inhibition of M. capricolum growth decreased the ability of the cells to take up exogenous phospholipids and cholesterol. The possibility that the contact between the lipid donors and the membrane involves specific receptors best exposed in actively growing cells is discussed.     

Phospholipids and glycolipids in subcellular fractions of mosquitoAedes aegypti cells

Summary Thin layer chromatography of [¹⁴C]palmitate labeled phospholipids of mosquitoAedes aegypti cells reveals that phosphatidylethanolamine is the major phospholipid, and ceramide phosphorylcholine the major sphingolipid. Glycolipids of these cells contain mannose in addition to glucose. The distribution of phospholipids in subcellular membrane fractions shows an enrichment in sphingomyelin and phosphatidyl serine in plasma membranes and other smooth membrane fractions. Cardiolipin is located predominantly in fractions, rich in mitochondrial membranes.     

Antigen-and ionophore-stimulated synthesis of platelet-activating factor by the cloned mast cell line, MC9

MC9 mast cells stimulated by a soluble (calcium ionophore A23187) or by an Fc epsilon-receptor agonist (IgE plus hapten) produce platelet activating factor (PAF). MC9 cells incorporate either exogenous [3H]acetic acid or [3H]lyso-PAF into PAF. PAF was identified by mobility on thin layer chromatography, platelet aggregatory activity inhibitable by known PAF antagonists, and by enzymatic modification. Quantified by aggregation of rabbit platelets, MC9 cells produce 6 pmoles PAF/10(6) cells. MC9 cells express acetyltransferase activity of 0.19 nmole/5 min-mg protein. Analysis of MC9 phospholipids by HPLC showed that MC9 cells contain large amounts of phosphatidylcholine (82 nmoles/10(7) cells) but contain little ether-linked phosphatidylcholine (4 nmoles/10(7) cells).     

Lipid synthesis by germinating soya bean

Lipid content and changes during the first 2 days of germination have been examined in soya bean Glycine max (L.) Merr variety Fiskeby V. Triacylglycerol, the principle storage lipid, is reduced on germination and this is accompanied by a rise in phospholipid content. The relative amounts of phospholipids rise equally, apart from the disappearance of N-acylphosphatidylethanolamine and significant increases in the proportions of phosphatidic acid and phosphatidylglycerol.Incorporation of acetate-[¹⁴C] into lipids occurs after a brief lag phase. Labelling is almost entirely (94–100%) confined to the acyl portion of the major complex lipids. Triacylglycerols had low specific radioactivities and, of the phospholipids, phosphatidylglycerol, cardiolipin, and phosphatidylinositol had the highest specific radioactivities. Incorporation was somewhat reduced by protein synthesis inhibitors and was mainly into palmitic, stearic and oleic acids. There were minor differences in their distribution within lipid types.     

Phospholipid synthesis and exchange in isolated liver cells

1. The [(32)P]phosphate incorporated into the phospholipids of isolated rat hepatic cells is present in phosphatidic acid and to a smaller extent in phosphatidylinositol. 2. The ability to synthesize nitrogen-containing phospholipids is restored by adding a liver supernatant fraction, and it is suggested that the metabolic deficiency is caused by the leakage of cytoplasmic enzymes of the synthetase system from the cells. 3. Fortified cell preparations were pulse-labelled with [(32)P]phosphate, [Me-(14)C]choline, [2-(14)C]ethanolamine and [U-(14)C]inositol and the subsequent fate of the labelled microsomal and mitochondrial phospholipids followed. 4. A fall in the specific radioactivity of microsomal phospholipids and a rise in that of mitochondrial phospholipids is interpreted as providing evidence of a transfer of labelled phospholipid molecules from the synthetic site (endoplasmic reticulum) to the mitochondrial membranes in the intact cells. 5. The formation of the phospholipids of mitochondrial membranes is discussed.     

Myosin synthesis by fusion-arrested chick embryo myoblasts in cell culture.

The synthesis and accumulation of myosin was studied in subcultures of fusion-blocked, postmitotic embryonic chicken myogenic cells. Electron micrographs and fluorescent microscopy with antimyosin revealed that most, if not all, of these cells contain myosin. It was also found that these cells are capable of accumulating myosin at rates comparable to fused cells. Incipient T-tubule formation was also present in some of the blocked cells. It is concluded that cell fusion is not a prerequisite for myosin synthesis and accumulation or T-tubule formation during myogenesis in vitro.     

The distribution of fatty acids in gill phospholipids of the Chinese crab Eriocheir sinensis

The fatty acid composition of the different classes of phospholipids isolated from 10,000 g fractions of posterior and anterior gills of fresh water-acclimatized Chinese crabs (Enocheir sinensis) has been analysed by two-dimensional TLC and GLC. All the phospholipids, especially PE and DPG, contain large amounts of long chain, polyunsaturated fatty acids. In PC and PE of both the anterior and posterior gills, polyenic fatty acids (particularly the ω-3-acid family) were found to be mainly incorporated in the 2-positon of the glycerol molecule. The ω-3-fatty acids/ω-6-fatty acids ratio in each class of phospholipids is higher in the posterior gills than in the anterior ones. The sum of polyunsaturated fatty acids and the unsaturation index of DPG are more important in the posterior gills than in the anterior ones. It is suggested that negatively charged unsaturated DPG and that the contrast of activities of ω-6- and ω-3-acids found in phospholipids provide a suitable lipid environment for optimal activity of the transport mechanisms at work in posterior gills of the Chinese crab.     

Phosphatidylethanolamine synthesis is required for optimal virulence of Brucella abortus

The Brucella cell envelope contains the zwitterionic phospholipids phosphatidylcholine (PC) and phosphatidylethanolamine (PE). Synthesis of PC occurs exclusively via the PC synthase pathway, implying that the pathogen depends on the choline synthesized by the host cell to form PC. Notably, PC is necessary to sustain a chronic infection process, which suggests that the membrane lipid content is relevant for Brucella virulence. In this study we investigated the first step of PE biosynthesis in B. abortus, which is catalyzed by phosphatidylserine synthase (PssA). Disruption of pssA abrogated the synthesis of PE without affecting the growth in rich complex medium. In minimal medium, however, the mutant required choline supplementation for growth, suggesting that at least PE or PC is necessary for Brucella viability. The absence of PE altered cell surface properties, but most importantly, it impaired several virulence traits of B. abortus, such as intracellular survival in both macrophages and HeLa cells, the maturation of the replicative Brucella-containing vacuole, and mouse colonization. These results suggest that membrane phospholipid composition is critical for the interaction of B. abortus with the host cell.     

Acyl chain and head group regulation of phospholipid catabolism in senescing carnation flowers

Microsomal membranes from the petals of senescing carnation (Dianthus caryophyllus L.) flowers contain phosphatidylcholine, phosphatidylethanolamine, phosphatidylglycerol, and phosphatidylinositol. These phospholipid classes decline essentially in parallel during natural senescence of the flower and when microsomal membranes isolated from young flowers are aged in vitro. However, measurements of changes in the endogenous molecular species composition of microsomal phospholipids during natural senescence of the flower petals and during in vitro aging of isolated membranes have indicated that the various molecular species of phospholipids have quite different susceptibilities to catabolism. Acyl chain composition and the nature of the head group are both determinants of their susceptibility to catabolism. As well, a comparison of the phospholipid catabolism data for naturally senesced membranes and for membranes aged in vitro suggests that the phospholipid composition of membranes is continuously altered during senescence by acyl chain desaturation and possibly retailoring so as to generate molecular species that are more prone to catabolism. The results collectively indicate that provision of particular molecular species of phospholipids with increased susceptibility to degradation contributes to enhanced phospholipid catabolism in the senescing carnation petal.     

Phospholipid Composition of Desulfovibrio Species

The phospholipids of Desulfovibrio desulfuricans, Norway strain, D. vulgaris, and D. gigas were examined in relationship to their qualitative and quantitative composition. D. desulfuricans and D. vulgaris exhibited an essentially identical phospholipid composition consisting of phosphatidylethanolamine, phosphatidylglycerol, cardiolipin, and lysophosphatidylserine. Phosphatidylserine (10.9%) was present in D. desulfuricans but was not detected in D. vulgaris. D. gigas was found to contain only two phospholipids, phosphatidylethanolamine (30%) and phosphatidylglycerol (70%). An ornithine-containing lipid was detected in D. gigas which was not present in the other two Desulfovibrio species.     

The de-repression of thiamine biosynthesis by adenosine. A tool for investigating this biosynthetic pathway

1. Growth of Salmonella typhimurium LT2 in the presence of adenosine was shown to cause enormous synthesis of thiamine in washed-cell suspensions. 2. Evidence that this was due to de-repression and not an accumulation of precursors was obtained by using a mutant blocked in the biosynthesis of the thiazole moiety, which showed a similarly large synthesis of the pyrimidine of thiamine. 3. The specific requirements for a source of energy, nitrogen and sulphur were investigated, and indicated new synthesis in this system.     

Heterogeneity of insect flight muscle mitochondria as demonstrated by different phospholipid turnover in vivo

Homogenates of the flight muscle of Periplaneta americana were separated by differential centrifugation into several fractions. Examination by electron microscopy indicated that two of these fractions contain predominantly intact mitochondria. When expressed relative to the protein content they have a high content of diphosphatidylglycerol and their phospholipids incorporate [³²P]-orthophosphate in vivo much more slowly than phospholipids in the supernatant. The mitochondria sedimenting at the lower speed have, in vivo, a significantly lower incorporation or radioactive phosphate into diphosphatidylglycerol and phosphatidylethanolamine than those of the lighter fraction. From developing muscle, fractions of similar composition and yield can be obtained, but they do not differ in the rate of incorporation of radioactive phosphate into diphosphatidylglycerol in vivo.     

Oxidized phospholipids: Biomarker for cardiovascular diseases

Biologically active oxidized phospholipids can initiate and modulate many of the cellular events attributed to inflammation leading to atherosclerosis. Produced by enzymatic or non-enzymatic processes, these molecules interact with various cells via specific receptors and in general give rise to inflammatory signals. There is considerable evidence that oxidized phospholipids accumulate in vivo and play significant roles in atherosclerosis and thrombosis, suggesting that oxidized phospholipids could be biomarkers that reflect the global extent of these diseases in vivo. Thus, understanding the biosynthetic pathways, receptor specificity and signaling processes of oxidized phospholipids is important in understanding atherosclerosis, thrombosis and related inflammatory diseases.