Lipid composition of plasma membranes from barley leaves and roots, spinach leaves and cauliflower inflorescences

Highly purified plasma membranes (PM) were obtained from barley (Hordeum vulgare L. cv. Kristina) leaves and roots, spinach (Spinacia oleracea L. cv. Viking II) leaves, and cauliflower (Brassica oleracea) inflorescences by partitioning in an aqueous polymer two-phase system. The sterol and polar lipid composition of the PM, including the fatty acid composition of the glycerolipids, was determined. Dominating lipids were free sterols, glucocerebroside, phosphatidylcholine (PC) and phosphatidylethanolamine (PE), although large variations in content were observed between the PM of the different species and organs. Thus, the spinach leaf PM contained only 7% (mol %) free sterol compared to over 30% free sterol in the other PM analysed, with the barley root PM as the other extreme (57% free sterol). On the other hand, sterol derivatives were more abundant in the spinach leaf PM, containing 13% acylated sterol glycosides. Cerebroside constituted 16% of the lipids in the barley leaf PM but only 3% in cauliflower. The phospholipids PC and PE ranged from 25 and 24%, respectively, in the spinach leaf PM to 8 and 7%, respectively, in the barley root PM. As a result of the large variations in sterol and phospholipid content, the ratio of free sterol to phospholipid varied from 2.2 in the barley root PM to only 0.1 in the spinach leaf PM. Sitosterol, campesterol and stigmasterol were the completely dominating sterols in the barley and cauliflower PM, whereas the unique sterol composition of spinach was dominated by spinasterol. Palmitic (16:0), linoleic (18:2) and linolenic (18:3) acid were the major glycerolipid fatty acids. The fatty acid composition of the barley root PM was the most saturated (44% 16:0, 13% 18:3), whereas that of the cauliflower PM was the most unsaturated (21% 16:0,42% 18:3). Thus, very large variations were observed in both total lipid and fatty acid composition of the PM investigated, which represent both mono— and dicotyledons, as well as both photosynthetic and non-photosynthetic tissue. The consequences of this large diversity in composition of the lipid bilayer for the function of integral PM proteins are discussed.     

Composition of phospholipids and phospholipid fatty acids in RAT mast cells

Summary The composition of phospholipids and phospholipid fatty acids in isolated rat serous fluid mast cells was analyzed by thin layer chromatography, gas-liquid chromatography and mass spectrometry. The phospholipids constituted about 50% of the mast cell lipids and phosphatidylethanolamine, phosphatidylserine, phosphatidylinositol, phosphatidylcholine, sphingomyelin and lysophosphatidylcholine were identified. The phosphatidylethanolamine fraction contained aldehydes and the highest proportion of unsaturated fatty acids. Sphingomyelin contained predominantly saturated fatty acids whereas the ratio unsaturated fatty acids: saturated fatty acids for the other phospholipids was more close to 1.     

Lipid and Fatty Acid composition of chloroplast envelope membranes from species with differing net photosynthesis

Lipid and fatty acid compositions were determined for chloroplast envelope membranes isolated from spinach (Spinacia oleracea L.), sunflower (Helianthus annuus L.), and maize (Zea mays L.) leaves. The lipid composition was similar in sunflower, spinach, and undifferentiated maize chloroplast envelope membranes and different in maize mesophyll chloroplast envelope membranes. The predominant lipid constituents in all envelope membranes were monogalactosyldiglyceride (27 to 46%), digalactosyldiglyceride (18 to 33%), and phosphatidylcholine (7 to 30%). The fatty acid composition was also similar in sunflower and spinach chloroplast envelope membranes in comparison to those from maize. The major acyl fatty acids of the chloroplast envelope membrane were palmitic (C_16:0, 41 and 36%) and linolenic (C_18:3, 29 and 40%) acids for spinach and sunflower; palmitic (77%) and stearic (C_18:0, 12%) acids for young maize; and palmitic (61%), stearic (14%), and linolenic (13%) acids for mature maize. The differences in lipid and acyl fatty acid compositions among these plants which vary in their rates of net photosynthesis were largely quantitative rather than qualitative.     

Fatty chain composition of phospholipids in sea urchin spermatozoa

1. An analysis was made of lipids extracted from the spermatozoa of the sea urchins, Hemicentrotus pulcherrimus and Anthocidaris crassispina. 2. Nearly all the lipids from both species consisted of phospholipids (about 80%) and cholesterol (about 14%). Triglyceride and cholesterol ester were present in trace amounts. 3. The fatty acid composition of each phospholipid was determined by gas-liquid chromatography. In both species, the fatty acid consisting of phosphatidylethanolamine was of the unsaturated type for the most part, while cardiolipin was comprised to a considerable degree of saturated fatty acids. In phosphatidylcholine and phosphatidylserine from H. pulcherrimus sperm, unsaturated fatty acid content was somewhat higher than that in phospholipids from A. crassispina sperm.     

Lipid composition and sphingolipid fatty acids of myelin in fruit bat brain

Myelin was isolated from the brain of adult fruit bats (Rousettus aegyptiacus) in a discontinuous sucrose gradient. Cholesterol comprised 189.0 mol/100 mol lipid phosphorus, galactolipids 60.3 mol/100 mol phosphorus and plasmalogens 32.5 mol/100 mol phosphorus. Choline and ethanolamine glycerophosphatide were present in nearly equal amounts followed by serine glycerophosphatide, sphingomyelin and inositol glycerophosphatides.The fatty acid composition of sphingomyelin and non-hydroxy cerebroside was determined by gas-liquid chromatography. Fatty acids were mainly saturated or mono-unsaturated with a small percentage of polyunsaturated fatty acids present.The lipid composition and sphingolipid fatty acid distribution in bat myelin was fairly similar to that of other species.     

Membrane perturbation by mastoparan 7 elicits a broad alteration in lipid composition of L1210 cells

Mastoparan 7 (Mas-7), an amphiphilic peptide possessing membrane perturbing activity, has been known to selectively stimulate some lipases. To examine changes in the lipid composition induced by Mas-7, we carried out systemic lipid analysis of L1210 cells after Mas-7 treatment. The total lipid was determined by HPLC, gas-liquid chromatography, and electrospray ionization mass spectrometry in conjunction with differential radiolabelling with [(32)P]orthophosphate, [(3)H]myristic acid, and [(3)H]arachidonic acid. The lipid analysis revealed multiple changes in more than 10 lipid classes. Free fatty acids (FFAs) and phosphatidylethanol (PEt), the phospholipase D product in the presence of ethanol, were increased significantly and phosphatidylcholine (PC) was decreased. Digitonin, a membrane permeabilizing reagent, similarly affected the lipid composition of L1210. The FFA released showed a very broad distribution of saturated, monounsaturated, and polyunsaturated fatty acids, implying that phospholipase A(2) alone could not account for all of the FFAs released. By comparing the molecular species of PEt with those of endogenous PC, we showed that phospholipase D in L1210 cells appeared to act selectively on diacyl-PC. The perturbation-induced alterations in the lipid composition brought about by Mas-7 might play a crucial role in the physiology of the affected cells.     

Homeoviscous adaptation under pressure. III. The fatty acid composition of liver mitochondrial phospholipids of deep-sea fish

Homeoviscous adaptation of biological membranes to high hydrostatic pressure has been investigated by determining the differences in lipid composition of membranes from fish obtained from depths between 200 and 4000 m. The fatty acid composition of phosphatidylcholine (PC), phosphatidylethanolamine (PE), phosphatidylserine/inositol and cardiolipin from a liver mitochondrial fraction was analysed by capillary gas-liquid chromatography. The ratio of saturated to unsaturated fatty acids significantly and negatively related to depth in PC and PE as predicted by homeoviscous adaptation to pressure. Thus, deep sea species possess greater proportions of unsaturated fatty acids than shallow species. Cardiolipin showed the opposite trend. An unsaturation index was not significantly related to depth in any phospholipid fraction.     

Characterization of the phospholipid and fatty acid composition of Sendai virus

The lipid composition of Sendai virus, propagated in chicken eggs, was analyzed by high performance liquid chromatography (HPLC), thin-layer chromatography (TLC), and gas-liquid chromatography (GLC). Phosphatidylcholine was found to be the dominant phospholipid (37.3%) with phosphatidylethanolamine (26.8%) and phosphatidylserine (12.0%) also present in significant amounts. Analysis of the fatty acid methyl esters revealed that the dominant fatty acids in total phospholipid were: C16:0 (17.6%), C18:0 (15.4%), C18:1 (n-9) (22.0%), and C24:0 (6.0%). Cardiolipin, phosphatidylserine, and sphingomyelin contained higher levels of saturated fatty acids relative to phosphatidylinositol, phosphatidylethanolamine, and phosphatidylcholine.     

FATTY ACIDS IN LOTIC PERIPHYTON: ANOTHER MEASURE OF COMMUNITY STRUCTURE1

The fatty acid spectra of 6 periphyton communities developed in laboratory streams at different combinations of light intensity and current velocity were determined by gas-liquid chromatography and silver nitrate thin-layer chromatography. Differences in species composition of the communities apparently had no striking effect on proportions of palmitic and stearic acids, whereas concentrations of myristic, palmitoleic, oleic, linoleic, and linolenic acids and a C_20:5 acid were more closely related to taxonomic differences. In general, communities dominated by blue-green algae exhibited relatively high proportions of oleic, linolenic, and linolenic acids and low proportions of palmitoleic acid and a C_20:5 acid, as compared to communities consisting primarily of diatoms. The data also indicated an inverse relationship between fatty acid redundancy and species diversity.     

Analysis of lipids in the salivary glands of Amblyomma americanum (L.): Detection of a high level of arachidonic acid

Analysis of lipids in salivary glands of the lone star tick, Amblyomma americanum, demonstrated that arachidonic acid (20:4, n-6) comprises 8% of all fatty acids identified by gas chromatography. The occurrence of arachidonic acid and other C₂₀ polyunsaturated fatty acids in tick salivary glands was confirmed by gas chromatography-mass spectrometry. Arachidonate is located entirely in the phospholipid fraction and is associated exclusively with phosphatidylcholine (PC) and phosphatidylethanolamine (PE). Salivary glands stored and frozen for several months had a similar lipid composition as freshly dissected salivary glands, with the exception of a small amount of free arachidonic acid and an increase in lysophosphatidylcholine. Incubation of salivary gland homogenates with snake venom phospholipase A₂ showed that most saturated fatty acids are esterified in the sn-1 position of PC and PE, with the unsaturated fatty acids in the sn-2 position. Approximately 75% of arachidonic acid is in the sn-2 position of PC and PE, adding support to the hypothesis that arachidonic acid is released into the cytoplasm after activation of a phospholipase A₂ for subsequent metabolism to prostaglandins and/or other eicosanoids. © 1993 Wiley-Liss, Inc.     

Gas-liquid chromatography of non-esterified fatty acids of the rat myocardium]

Non-esterified fatty acids in cardiac cell cultures of postnatal rat, in postnatal rat heart and in ventricle muscle of adult rat were analyzed by gas-liquid chromatography. Among cultured heart cells, three different types were found. Type A0 appeared when arachidonic acid was lacking in tissue culture medium; it had the highest percentage in saturated fatty acids, palmitoleic and linolenic acids. Type A1 and type A2 appeared when arachidonic acid was proved to be present in culture medium. Non esterified fatty acid composition of type A1 was closely similar to that of postnatal rat heart, while type A2 ranged between that of postnatal heart and that of adult ventricular muscle.     

Positional distribution of fatty acids in glycerophosphatides of bovine gray matter

Glycerophosphatides were isolated from ox brain gray matter by column chromatography. The fatty acid compositions of ethanolamine glycerophosphatides (EGP), serine glycerophosphatides (SGP), and choline glycerophosphatides (CGP) were determined by gas-liquid chromatography. The positional distribution of fatty acids in these glycerophosphatides were determined by phospholipase A hydrolysis (Habu habu venom). C(20) and C(22) polyunsaturated acids were confined almost exclusively to the 2-position of these lipids, where they comprised the majority of 2-substituents in EGP and SGP (oleic acid predominated in this position in CGP). In the 1-position, palmitoyl was the major substituent in CGP, stearoyl in SGP, and stearoyl or the corresponding alk-1-enyl group in EGP.     

Fatty acid composition of mitochondrial phospholipids of the ischemic heart

Gas liquid chromatography was used to study fatty acid (FA) composition of lysophosphatidylcholine (lyso-PC) and phosphatidylcholine in isolated rabbit heart mitochondria. In control, lyso-PC and PC were found to contain 95 and 66% of unsaturated FA, respectively. At 1 hour of ischemia (autolysis at 37 degrees C) the percentage of saturated FA in lyso-PC noticeably increased whereas FA composition remained unchanged. It is concluded that changes in FA composition of lyso-PC are caused by phospholipase A2 action.     

The structure and biosynthesis of phosphatidyl inositol in cauliflower inflorescence

Phosphatidyl inositol has been isolated from cauliflower inflorescence and soybean. Analysis of the fatty acid distribution showed that unsaturated fatty acids were preferred at position 2, and that saturated fatty acids were preferred at position 1.     

THE BIOCHEMICAL ANALYSIS OF SOME ESTUARINE PHYTOPLANKTON SPECIES. I. FATTY ACID COMPOSITION12

The fatty acid composition of 10 species of estuarine phytoplankton was determined using gas-liquid chromatography. Nine of the species were isolated from Yaquina Bay, Oregon. These species were common components of the phytoplankton of the bay. The tenth species, Isochrysis galbana, was obtained from the Culture Collection of Algae at Indiana University. The 10 organisms comprised 3 species of Chlorophyta, 1 species of Cryptophyta, and 6 species of Chrysophyta. Twenty-eight fatty acids were found in the 10 species. The fatty acids common to all species were 14:0, 16:0, 16:1, 13:0, 18:1, and 18:2. The Chlorophyta were distinguished by a high content of linolenic acid, 18:3. The diatoms were distinguished by very low amounts of C₁₈ acids and high proportion of 20:5. All species contained a high proportion of palmitic acid, 16:0.     

Disaturated and dipolyunsaturated phospholipids in the bovine retinal rod outer segment disk membrane.

Thin-layer chromatography was used to separate the major phospholipid headgroup classes of the rod outer segment disk membrane into subfractions which differ markedly in fatty acid composition. At least 18% of the rod outer segment phosphatidylcholine must contain two saturated fatty acids. Furthermore, two unsaturated fatty acids are found in at least 43% of the phosphatidylserine, 24% of the phosphatidylcholine, and 24% of the phosphatidylethanolamine. The unsaturated acids are predominantly polyunsaturated in all cases. A similar separation, but with less resolution, was achieved with silicic acid column chromatography. The temperature dependence of the polarization of the fluorescence of trans-parinaric acid (9,11,13,15-all-trans-octadecatetraenoic acid) showed that the thermal behavior of aqueous dispersions of the phosphatidylcholine subfractions was consistent with their fatty acid compositions.     

Fatty acid composition of cholesterol esters in brains of patients with Schilder's disease, G M1 -gangliosidosis and Tay-Sachs disease, and its possible relationship to the -position fatty acids of lecithin

Abstract— Cholesterol esters were isolated from cerebral cortex and white matter of patients with Schilder's disease, G_M1-gangliosidosis and Tay-Sachs disease, and the fatty acid composition was determined by gas-liquid chromatography. The fatty acid composition was similar among the three pathological conditions, but it was entirely different from that reported for cholesterol esters of normal brain. Lecithin and ethanolamine phospholipids were isolated from the same brain specimens, treated with snake venom phospholipase A, and the fatty acids at the a’and β-positions of the glycerol moiety were determined separately. The fatty acid composition of cholesterol esters was similar to that of the β-position fatty acids of lecithin of white matter in all samples, and was quite different from those of the a'-position of lecithin, or of the a’or β-position of ethanolamine phospholipids. The results indicate that the source of fatty acids for cholesterol esterification in nonspecific sudanophilic demyelination is different from that in normal brain, and that the most likely source is the β-linked fatty acids of lecithin. There are two possible enzymic mechanisms; activation of phospholipase A and subsequent esterification of the liberated β-position fatty acids to cholesterol, or direct transacylation by lecithin-cholesterol acyl transferase.     

Phospholipid and fatty acid composition in mitochondria from spinach (Spinacia oleracea) leaves and petioles. A comparative study.

Essentially chlorophyll-free mitochondria from photosynthetic (leaf) and non-photosynthetic tissue (petiole) were isolated from spinach (Spinacia oleracea). Leaf mitochondria were found to contain more phosphatidylcholine than phosphatidylethanolamine compared with petiole mitochondria. Galactolipids were found in small and equal amounts (5 mol of galactolipids/100 mol of galactolipids and phospholipids) in both leaf and petiole mitochondria. Fatty acid composition showed a significant difference in the amounts of C18:2 and C18:3 acids. The C18:2/C18:3 ratio was more than twice as high in all of the phospholipids studied from petiole mitochondria compared with the ratio in leaf mitochondria. More than 50% (mol/100 mol) of the fatty acids in the major lipids (phosphatidylcholine, phosphatidylethanolamine and cardiolipin) in petiole mitochondria were C18:2. In the minor lipids (phosphatidylinositol and phosphatidylglycerol), C16:0 dominated in both leaf and petiole mitochondria.     

Energy metabolism of sea urchin spermatozoa, with phosphatidylcholine as the preferred substrate

Phosphatidylcholine content in the spermatozoa of the sea urchin, Hemicentrotus pulcherrimus, decreased rapidly during incubation with sea water. Sea urchin sperm contained approx. 85% phospholipid in total lipid. Phosphatidylcholine (PC) was the principal lipid. Other phospholipids, however, remained at constant levels during incubation. Although the free fatty acid content gradually increased following dilution of dry sperm in sea water, the amounts of triacylglycerol and cholesterol ester were extremely low. Analysis by gas-liquid chromatography indicated most of fatty acid moieties in PC to be polyenoic. PC composed in part of unsaturated fatty acids was consumed to a greater extent during incubation than that consisting of saturated fatty acids. Furthermore, 1-palmitoyl-2-[1-14C]linoleoylphosphatidylcholine was transformed to 14C-labelled free fatty acid in a subcellular system. Thus, possibly, phospholipase A2 is present in sea urchin sperm. Also, [1-14C]oleic acid was immediately oxidized to 14CO2 by sperm. It is thus concluded that sea urchin sperm use phosphatidylcholine as a substrate for energy metabolism.     

Phospholipids of the differentiating species of Tetrahymena

The whole-cell phospholipid composition of the six known polymorphic species of Tetrahymena has been examined by [(3)H]acetate and [(3)H]myristic acid radiolabeling, and by gas-liquid chromatography of total phospholipid-bound fatty acids. Five of the polymorphic species contained similar phospholipid profiles following radiolabeling in that phosphatidylethanolamine (PE) was the predominant phospholipid; however, in cells of Tetrahymena patula LFF, aminoethylphosphonolipid was present in amounts nearly equal to PE. Tetrahymena patula LFF contained an unusually large percentage of sphingolipid (16.2% by [(3)H]acetate radiolabeling). Substantial differences were found in the fatty acid profiles of the polymorphic species, which included the degree of fatty acid unsaturation and relative weight percentages of odd-chain fatty acids. Tetrahymena vorax contained a low ratio of unsaturated C(18) fatty acids to saturated C(18) fatty acids as compared with all other species examined. The differentiating species generally contained a lesser percentage of monoenoic fatty acids and a lower ratio of unsaturated C(16) fatty acids to saturated C(16) fatty acids as compared with the two monomorphic species examined.