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PubMed1992

The effect of borage oil consumption on the composition of individual phospholipids in human platelets.

The effect of supplementation with borage oil containing gamma-linolenic acid (GLA, 18:3n-6) on the levels and fatty acid compositions of individual human platelet phospholipids was evaluated. For this purpose, male volunteers were given an average daily intake of 5.23 g of GLA (as borage oil) for 42 days, after which the supplement was withdrawn for an additional 42-day period. No significant differences were found in the relative amounts of the choline phospholipids (PC), ethanolamine phospholipids (PE), phosphatidylserine (PS), phosphatidylinositol (PI), and sphingomyelin (SPH) at days 0, 22, 43, 64, and 85. However, marked differences were observed in the fatty acid compositions of all the phospholipids including a marked, and reversible, rise in the level of dihomo-gamma-linolenic acid (DGLA, 20:3n-6), without a significant elevation in arachidonic acid (AA, 20:4n-6) and decreases in n-3 polyunsaturated fatty acids. In the case of PC, a net rise in DGLA of 1.8 mol% was observed by day 22 (from 2.1 to 3.9 mol%). The DGLA/AA ratios at day 43 exhibited considerable variability across phospholipids with PC greater than PS greater than PE = PI; the PC, PE, PS, and PI accounted for 67.6, 16.7, 12.9, and 2.6%, respectively, of the total DGLA in platelet phospholipids. Interestingly, despite the lack of DGLA in SPH, this phospholipid exhibited a marked enrichment in nervonic acid (NA, 24:1n-9) from 16.2 to 24.7 mol% upon borage oil consumption. The observed alterations may represent biochemical strategies for adaptation to dietary fatty acid modifications and the regulation of platelet membrane functioning.

Adult↗
PubMed1993

Phospholipids in Drosophila heads: effects of visual mutants and phototransduction manipulations.

A procedure was developed to label phospholipids in Drosophila heads by feeding radioactive phosphate (32Pi). High-performance thin-layer chromatography showed label incorporation into various phospholipids. After 24 h of feeding, major phospholipids labeled were phosphatidylethanolamine (PE), 47%; phosphatidylcholine (PC), 24%; and phosphatidylinositol (PI), 12%. Drosophila heads have virtually no sphingomyelin as compared with mammalian tissues. Notable label was in ethanolamine plasmalogen, lysophosphatidylethanolamine, lysophosphatidylcholine and lysophosphatidylinositol. Less than 1% of the total label was in phosphatidylinositol 4-phosphate and phosphatidylinositol 4,5-bisphosphate. Other lipids labeled included phosphatidylserine, phosphatidic acid and some unidentified lipids. A time course (3-36 h) study revealed a gradual decrease in proportion of labeled PI, an increase in proportion of labeled PC and no obvious change in labeled PE. There were no significant differences in phospholipid labeling comparing the no receptor potential (norpA) visual mutant and wild type under light vs. dark conditions. However, overall 32P labeling was higher in the wild type fed in the light as compared to the dark and to norpA either in light or dark. This suggests that functional vision facilitates incorporation of label. Differences in phospholipid labeling were observed between young and aged flies, particularly in lysophospholipids and poly-PI, implicating phospholipase A2 function in recycling. v Manipulations such as the outer rhabdomeres absent and eyes absent mutants and carotenoid deprivation failed to yield notable differences in phospholipid labeling pattern, suggesting that phospholipids important to vision may constitute only a minor portion of the total labeled pool in the head.

Animals↗
PubMed1994

Meningioma phospholipid profiles measured by 31P nuclear magnetic resonance spectroscopy.

Fourteen cases of intracranial meningioma were characterized after chloroform/methanol extraction by 31P nuclear magnetic resonance (NMR) spectroscopy at 202.4 MHz. Each phospholipid class detected in the extracts was identified and quantitated in terms of its molar percentage relative to the total phospholipids measured. The following phospholipids were assayed by 31P NMR: phosphatidylglycerol, phosphatidic acid, diphosphatidylglycerol, ethanolamine plasmalogen, phosphatidylethanolamine (PE), lysophosphatidylinositol, phosphatidylserine, sphingomyelin, lysophosphatidylcholine (LPC), phosphatidylinositol (PI), sphingosylphosphorylcholine and phosphatidylcholine. In addition, two unidentified phospholipids were detected with resonances at 0.13 and -0.78 ppm, respectively. Three distinct types of spectra were obtained on the extracts and grouped accordingly for comparison purposes. Type 1 tumors showed unusual 31P NMR profiles with low levels of PE and PI and elevated levels of LPC; type 2 tumors were characterized by low levels of the ethanolamine phospholipids and near equivalent levels of PI and LPC. The spectra of type 1 and type 2 tumors were characteristic of degenerative cells that lacked membrane permeability associated with loss of ethanolamine plasmalogen in the presence of significant phospholipid turnover. Meningiomas belonging to the third spectral type showed characteristics similar to those of normal tissues with normal levels of PE and ethanolamine plasmalogen, as well as very low levels of LPC relative to PI. Type 3 tumors lacked the characteristic signs of degeneration noted in type 1 and type 2 tumors. The data corroborate and augment in vivo spectroscopic findings reported earlier and demonstrate the value of 31P NMR spectroscopic phospholipid analysis on lipid extracts for the characterization of meningiomas.

Humans↗
PubMed2004

Phospholipids reduce the intraperitoneal adhesion of colonic tumor cells in rats and adhesion on extracellular matrix in vitro.

OBJECTIVE: To evaluate the influence of intraperitoneal treatment with phospholipids on the formation of peritoneal carcinosis after inoculation of colonic tumor cells in rats. The presence of tumor cells in the peritoneal cavity serves as a prognostic marker for postoperative survival after resection of gastrointestinal cancer. Intraperitoneal tumor cell attachment is a pivotal step in developing peritoneal carcinosis. Intraabdominal application of phospholipids resulted in a significant decrease of adhesion formation, especially at sites of peritoneal lesions. METHODS: 2x10(6) colonic tumor cells (DHD/K12/TRb) were injected intraperitonely in female BD-IX rats. A total of 90 rats were divided into three groups with treatments of phospholipids at 75 mg/kg or 150 mg/kg bodyweight or sodium chloride at 0.9% in the control group. The treatment groups were subdivided into animals with defined peritoneal lesions and animals without lesions. After 30 days, the extent of peritoneal carcinosis was determined by measuring the tumor volume, the area of tumor attachment and the Peritoneal Cancer Index. Over a 90-day observation period, the survival rate was analyzed. In vitro, we examined the reduction of tumor cell adhesion on extracellular matrix components after treatment with phospholipids. Microtiter plates were coated with laminin, fibronectin or collagen IV for adhesion experiments. RESULTS: In our study, we found a significant reduction of peritoneal dissemination with respect to all evaluation methods after treatment with phospholipids at 150 mg/kg in animals without peritoneal lesions. This could not be achieved using the lower concentration of phospholipids (75 mg/kg). In vitro, the maximum reductions of tumor cell adhesion by phospholipids compared with the control values for laminin and fibronectin were 46% and 37%, respectively, whereas for collagen IV the reduction was only 24% ( p<0.0001). CONCLUSIONS: A new method of prevention of intraperitoneal tumor cell adhesion, possibly leading to a reduced incidence of peritoneal carcinosis after surgery of gastrointestinal tumors, is introduced.

Adenocarcinoma↗
PubMed1999

Effect of endurance training on the phospholipid content of skeletal muscles in the rat.

Only few data are available on the effect of training on phospholipid metabolism in skeletal muscles. The aim of the present study was to examine the effect of 6 weeks of endurance training on the content of particular phospholipid fractions and on the incorporation of blood-borne [14C]-palmitic acid into the phospholipids in different skeletal muscles (white and red sections of the gastrocnemius, the soleus and the diaphragm) of the rat. Lipids were extracted from the muscles and separated using thin-layer chromatography into the following fractions: sphingomyelin, phosphatidylcholine, phosphatidylserine, phosphatidylinositol, phosphatidylethanolamine, cardiolipin and neutral lipids (this fraction being composed mostly of triacylglycerols). It was found that training did not affect the content of any phospholipid fraction in soleus muscle. It increased the content of sphingomyelin in white gastrocnemius muscle, cardiolipin and phosphatidylethanolamine in red gastrocnemius muscle and phosphatidylinositol in white gastrocnemius muscle and diaphragm. The total phospholipid content in red gastrocnemius muscle of the trained group was higher than in the control group. Training reduced the specific activity of sphingomyelin and cardiolipin in all muscles, phosphatidylcholine in soleus, red, and white gastrocnemius muscles, phosphatidylserine in all muscles, phosphatidylinositol in all except the soleus muscle, and phosphatidylethanolamine in hindleg muscles, but not in the diaphragm compared to the corresponding values in the sedentary group. It was concluded that endurance training affects skeletal muscle phospholipid content and the rate of incorporation of the blood-borne [14C]palmitic acid into the phospholipid moieties.

Animals↗
PubMed2004

Lipids rich in phosphatidylethanolamine from natural gas-utilizing bacteria reduce plasma cholesterol and classes of phospholipids: a comparison with soybean oil.

We compared the effects of three different high-lipid diets on plasma lipoproteins and phospholipids in mink (Mustela vison). The 18 mink studied were fed one of the three diets during a 25-d period in a parallel group design. The compared diets had 0, 17, and 67% extracted lipids from natural gas-utilizing bacteria (LNGB), which were rich in PE. The group with 0% LNGB was fed a diet for which the lipid content was 100% soybean oil. The total cholesterol, LDL cholesterol, and HDL cholesterol of animals consuming a diet with 67% LNGB (67LNGB-diet), were significantly lowered by 35, 49, and 29%, respectively, and unesterified cholesterol increased by 17% compared with the animals fed a diet of 100% lipids from soybean oil (SB-diet). In addition, the ratio of LDL cholesterol to HDL cholesterol was 27% lower in mink fed the 67LNGB-diet than those fed the SB-diet. When the mink were fed the 67LNGB-diet, plasma PC, total phospholipids, lysoPC, and PI were lowered significantly compared with the mink fed a SB-diet. Plasma total cholesterol was correlated with total phospholipids as well as with PC (R = 0.8, P< 0.001). A significantly higher fecal excretion of unesterified cholesterol, cholesteryl ester, PC, lysoPC, and PE was observed in the 67LNGB-fed mink compared with the SB-fed mink. We conclude that phospholipids from the 67LNGB-diet decreased plasma lipoprotein levels, the LDL/HDL cholesterol ratio, and plasma phospholipid levels, especially lysoPC and PC, compared with the highly unsaturated soybean oil. Our findings indicate that the decrease of plasma cholesterol is mainly caused by a specific mixture of phospholipids containing a high level of PE, and not by the dietary FA composition. The lack of significant differences in the level of plasma PE due to the diets indicates that most of the PE from LNGB has been converted to PC in the liver. Thus, plasma cholesterol may at least be partly regulated by phospholipid methylation from PE to PC in the liver.

Animals↗
PubMed1998

Alterations in heart and kidney membrane phospholipids in hypertension as observed by 31P nuclear magnetic resonance.

Abnormalities of phospholipids in hypertension have previously been described in human erythrocyte, platelet, and plasma lipoproteins. Since the heart and kidney are adversely affected by hypertension, we investigated possible alterations in their membrane phospholipids, which could play a role in the derangement of intracellular ion balance widely observed in hypertension. The phospholipid compositions of heart and kidney from spontaneously hypertensive rats (SHR) and Wistar-Kyoto (WKY) rats were determined by using 31P nuclear magnetic resonance (NMR) spectroscopy. Absolute contents of all phospholipids in hypertensive hearts and kidneys were significantly higher than in normotensive hearts and kidneys. Expressed as a fraction of total phospholipid, cardiolipin (CL) and phosphatidylethanolamine plasmalogen (PEp) were significantly increased in SHR hearts compared to WKY hearts (CL and PEp were 7.95+/-0.22% and 13.16+/-0.35% in SHR vs. 7.01+/-0.20% and 11.19+/-0.42% in WKY rats, P< or =0.05), but phosphatidylethanolamine (PE) and phosphatidylcholine (PC) were significantly decreased in SHR (PE and PC were 22.46+/-0.37% and 44.81+/-0.43% in SHR vs. 24.02+/-0.44% and 46.01+/-0.50% in WKY rats, P< or =0.05). In the phospholipids extracted from rat kidneys, the percentage of PE was significantly higher for SHR than for WKY rats (20.37+/-0.60% vs. 18.43+/-0.37%, P< or =0.05), while PEp and phosphatidylserine (PS) were significantly lower for SHR (PEp and PS were 10.22+/-0.36% and 8.42+/-0.28% in SHRs vs. 11.29+/-0.36% and 9.71+/-0.40% in WKY rats, P< or =0.05). The above alterations in phospholipid composition might contribute to the higher oxygen consumption in the hypertensive heart and abnormal intracellular ion concentrations and ion transport in the heart and the kidney in hypertension.

Animals↗
PubMed1999

Positional analysis of triglycerides and phospholipids rich in long-chain polyunsaturated fatty acids.

Four sources of long-chain polyunsaturated fatty acids (LCP) differing in their chemical structure (triglycerides or phospholipids) and in their origin (tuna triglycerides, fungal triglycerides, egg phospholipids, and pig brain phospholipids) were analyzed to determine the distribution of the component fatty acids within the molecule. Lipase and phospholipase A2 hydrolysis was performed to obtain 2-monoacylglycerols and lysophospholipids, respectively, which allowed us to determine the distribution of fatty acids between the sn-2 and sn-1,3 positions of triglycerides or between the sn-1 and sn-2 position of phospholipids. Fatty acids in the LCP sources analyzed were not randomly distributed. In tuna triglycerides, half of the total amount of 22:6n-3 was located at the sn-2 position (49.52%). In fungal triglycerides, 16:0 and 18:0 were esterified to the sn-1,3 (92.22% and 91.91%, respectively)18:1 and 18:2 to the sn-2 position (59.77% and 62.62%, respectively), and 45% of 20:3n-6 and only 21.64% of 20:4n-6 were found at the sn-2 position. In the lipid sources containing phospholipids, LCP were mainly esterified to the phosphatidylethanolamine fraction. In egg phospholipids, most of 20:4n-6 (5.50%, sn-2 vs. 0.91%, sn-1) and 22:6n-3 (2.89 vs. 0.28%) were located at the sn-2 position. In pig brain phospholipids, 22:6n-3 was also esterified to the sn-2 (13.20 vs. 0.27%), whereas 20:4n-6 was distributed between the two positions (12.35 vs. 5.86%). These results show a different fatty acid composition and distribution of dietary LCP sources, which may affect the absorption, distribution, and tissue uptake of LCP, and should be taken into account when supplementing infant formulas.

Animals↗
PubMed1986

Micellization and solubilization of phospholipids by surfactants.

Phospholipids comprise the major component that defines biological membranes. Yet these amphiphilic molecules exist as liquid crystals when they are dispersed in water. Surfactants which form micelles in aqueous solution are quite useful in solubilizing the phospholipids by converting them into mixed micelles. Phospholipids containing short-chain fatty acids have also been chemically synthesized and these form micelles themselves without added surfactants. Below their cmc, monomers of these synthetic phospholipids can be co-micellized with surfactants. The solubilization of natural phospholipids and the micellization of synthetic phospholipids as well as their co-micellization by surfactants will be discussed. Emphasis will be placed on nonionic surfactants where the formation of micelles and mixed micelles with phospholipids has been well studied.

Colloids↗
PubMed1991

Phospholipids at the oil/water interface: adsorption and interfacial phenomena in an electric field.

Interfacial effects produced in an immiscible liquid system by the action of an external electric field have been considered. The addition of small amounts of neutral phospholipids to the nonaqueous phase has been shown to result in a marked increase in the sensitivity of the interfacial boundary to the voltage applied, which is manifested by: (i) an accelerated decrease of the interfacial tension after the two immiscible liquid phases have been brought into contact; (ii) reduced interfacial tension, by 20-30 mN/m, at the oil/water interface at field strengths of 1-10 kV/m (the interfacial tension drop in the absence of phospholipids does not exceed 5 mN/m); (iii) development of electrohydrodynamic instability at the planar dividing surface between phases; and (iv) dispersion of water into the nonaqueous phase at smaller field strengths by a factor of about 100 as compared to those normally required in the absence of phospholipids. In order to gain a deeper insight into the mechanisms of interfacial phenomena, mainly exemplified by the n-heptane/water system containing phosphatidylcholine, three major issues have been considered: (1) Kinetics of the adsorption of phospholipid at the oil/water interface from the nonaqueous phase, and effects produced by exposure to an external electric field; also, the adsorption under equilibrium conditions, and the structure of the adsorption layer formed. (2) Interactions between neutral phospholipid and inorganic or organic ions at the interfacial boundary under the voltage applied. (3) Conditions for the occurrence of electrohydrodynamic instability at the dividing surface between oil and water and the formation of a water-in-oil emulsion; also aggregation and gelation processes induced in the nonaqueous phospholipid solution bulk by the action of a weak external electric field. Throughout the present paper, an attempt has been made to relate the microscopic behaviour of phospholipids under an external electric field to macroscopically observable properties at the movable interfacial boundaries. The adsorption studies have shown that phosphatidylcholine is prone to self-organization into a liquid-crystalline state at an immiscible liquid interface. The disintegration of the interfacial lipid film thus formed by the action of a weak electric field has been explained as due to an enhanced electrohydrodynamic instability of liquid crystals. This results in the formation of either an emulsion, or a microemulsion in the nonaqueous solution bulk. The formation of a microemulsion is manifested by the appearance of an optically anisotropic gel, stable only under an external applied electric field, in the nonaqueous solution bulk.(ABSTRACT TRUNCATED AT 400 WORDS)

Adsorption↗
PubMed1990

Preparative high-performance liquid chromatography purification of polyunsaturated phospholipids and characterization using ultraviolet derivative spectroscopy.

A preparative reversed-phase HPLC system utilizing an isocratic mobile phase to purify up to 10-mg quantities of phospholipids is described. The method was developed to separate oxidation products of polyunsaturated phospholipids from intact, parent lipids. The method is useful for phosphatidylcholine and phosphatidylethanolamine on a preparative scale and for phosphatidylserine and phosphatidic acid on an analytical scale. Both intact phospholipids and oxidized phospholipids were monitored by absorbance at 206 nm. The oxidation products were simultaneously monitored at 234 nm where the intact phospholipids have only a very slight end absorption. Second-derivative uv spectroscopy proved to be extremely useful to identify the presence or to verify the absence of oxidation products in phospholipid samples. For autoxidized docosahexaenoic acid containing phospholipids, the absorbance maximum of diene oxidation products is 237 nm for the trans,trans (t,t) isomer and 246 nm for the cis,trans (c,t) isomers. Similarly, five classes of triene oxidation product stereoisomers have distinct absorbance maxima detected by second-derivative spectroscopy ranging from 269 to 292 nm.

Chromatography, High Pressure Liquid↗
PubMed1983

Differential effects of phospholipids on skeletal alkaline phosphatase activity in extracts, in situ and in circulation.

Human skeletal alkaline phosphatase (ALP) purified from human bone was subject to competitive inhibitions by phospholipids including cephalins, lecithins, and phosphatidylinositol. Ki values ranged from 0.7 to 1.5 mM, at pH 9.5. As previously shown, the enzyme was subject to uncompetitive inhibition by imidazole. The inhibitory phospholipids potentiated this effect, and altered the nature of the imidazole inhibition, from uncompetitive to mixed type, suggesting that imidazole was bound more efficiently to the enzyme-phospholipid complex than to the enzyme-substrate complex. No interactions were observed between phospholipids and other uncompetitive inhibitors of ALP. The skeletal ALP activity of cultured chick calvarial cells was assayed both in situ and in extracts. Like the extracted human isoenzyme, the extracted chick ALP was subject to competitive inhibition by cephalin (Ki = 0.3 mM at pH 9.3) and an inhibitory interaction between cephalin and imidazole, but the same isoenzyme showed neither effect in situ. The value of Km,PNPP at pH 9.5 for chick skeletal ALP was 1.5 mM in extracts and 7.1 mM in situ. When embryonic chick bones were cultured in vitro, skeletal ALP activity was released into the serum-free medium. Unlike the same isoenzyme extracted from the bones, the ALP activity in the medium was not inhibited by cephalin and showed no inhibitory interaction between cephalin and imidazole. Similarly, human serum ALP activities were not as sensitive to phospholipid inhibition as the same isoenzymes extracted from tissues. Human skeletal ALP extracted from bone was inhibited by cephalin, but the skeletal isoenzyme in Pagetic serum was not, suggesting that the potential for phospholipid interaction was altered during or after release from osteoblast cell membranes. The observation that extracted human skeletal ALP lost its potential for inhibition by phospholipids after treatment with phospholipase C further suggests that ALP activity may be released from cells during membrane turnover.

Alkaline Phosphatase↗
PubMed1984

High-performance liquid chromatographic determination of phospholipid peroxidation products of rat liver after carbon tetrachloride administration.

A method to detect and determine phospholipid peroxidation products in a biological system was developed using reversed-phase high performance liquid chromatography and normal-phase HPLC. Reversed-phase HPLC could separate phosphatidylcholine (PC) hydroperoxides and phosphatidylethanolamine (PE) hydroperoxides of rat liver from the respective phospholipids. A linear relationship was observed between these hydroperoxides and their peak areas on the chromatogram. In the experiment with rats administered CCl4, reversed-phase HPLC gave prominent, large peaks attributable to the peroxidation of phospholipids, and the peroxide level of the liver phospholipids was tentatively determined. Normal-phase HPLC analysis confirmed that both PC and PE in the liver phospholipids were peroxidized after CCl4 treatment. Neither the thiobarbituric acid value of the liver homogenate nor the fatty acid composition of the liver phospholipid fraction showed any significant difference between CCl4-treated and control rats. It is concluded that normal-phase HPLC and reversed-phase HPLC can complement each other to serve as a direct and sensitive method for the determination of lipid peroxide levels in a biological source. However, it was difficult to distinguish phospholipid hydroperoxides from their hydroxy derivatives.

Animals↗
PubMed1986

Kinetic aspects of the role of phospholipids in D-beta-hydroxybutyrate dehydrogenase activity.

Interactions of phospholipids with D-beta-hydroxybutyrate dehydrogenase (BDH), a lecithin-requiring enzyme, have been studied by a kinetic approach. The process of reactivation of BDH by phospholipids, which follows a second-order mechanism, reveals that (1) at least 2 mol of lecithins is essential for the reactivation of the enzyme, and (2) the enzyme contains two dependent binding sites for lecithins. The graphic representation of the time course of reactivation shows a latent phase which decreases when there is an increase in the amount of phospholipids. A Scatchard plot treatment of the reactivation kinetic data reveals the presence of two classes of phospholipid binding sites, which exhibit high and low affinities related to the binding of four and two lecithin molecules, respectively. The effect of temperature on BDH activity and on the inactivation of the apoenzyme with N,N'-dicyclohexylcarbodiimide (a specific carboxyl reagent) or with phenylglyoxal (a specific arginine reagent) shows a break at 22-24 degrees C, indicating a slight structural change in the enzyme-active site around this temperature. In addition, the variations in enzyme kinetic parameters, according to the nature of phospholipids, are in agreement with conformational changes related to the nature and to the fluidity state of phospholipids. However, the apparent NAD+ binding constant does not depend on the phospholipid's fluidity.

Animals↗
PubMed1996

Phospholipid profiles in the oral yeast Candida.

Individual molecular species of phospholipids can now be readily detected using fast atom-bombardment mass spectrometry (FAB MS). The aim was to obtain detailed information on individual molecular species of Candida phospholipids using FAB MS. Lipids were extracted from Candida isolates and prepared for FAB MS analysis in negative-ion mode. Each isolate yielded several hundred anion peaks. The major anion peaks consistent with the presence of phospholipids included those of mass/charge (m/z) 836, 743, 715, 744 and 834 tentatively identified as phosphatidylethanolamine PE(43:5), phosphatidylglycerol PG(34:3), PG(32:3), PE(36:1) and PE(43:5). Major peaks consistent with the presence of carboxylate anions were of m/z 241, 253, 255, 277, 279, 281, 283, and 307 putatively identified as C15:0, C16:1, C18:3, C18:2, C18:1, C18:0 and C20:2 which also support the putative identifications of phospholipids. Such phospholipid profiles differ from those published for other microorganisms analysed by FAB MS. Quantitative differences were observed between different candidal species. Candida albicans may be readily differentiated from C. glabrata on the basis of its carboxylate anions. Within species, there are quantitative differences in phospholipid profiles. Thus, Candida has a unique combination of phospholipid analogues with potential chemosystematic significance.

Anions↗
PubMed1981

Regulation of phospholipid-ATPase complex interaction by the adenine nucleotide carrier.

(1) The effect of phospholipids on a preparation containing the ATPase complex and the adenine nucleotide carrier is studied in the presence of ligands known to affect the conformation of these components of the mitochondrial inner membrane. (2) When ATPase activity is abolished by phospholipid depletion, the reactivation induced by phosphatidylcholine is prevented by the simultaneous addition of ATP. ADP partially reproduces the ATP effect. AMP, GTP, UTP, and Pi are ineffective. (3) The influence of ATP is associated with reduced phospholipid binding to the membrane fragments and is reversible. The ATP effect on reconstitution is not manifest when phosphatidylcholine is added together with negatively charged phospholipids. (4) Carboxyatractyloside does not modify the phospholipid-ATPase complex interaction but bongkrekic acid is as effective as ATP. In the presence of ADP, the influence of bongkrekic acid is considerably increased. (5) It is concluded that the binding of ATP to the adenine nucleotide carrier enables the complex to select between the charged and uncharged phospholipids. As a result of the carrier conformational change, the ATPase complex is induced to prefer a negatively charged phospholipid environment.

Adenosine Triphosphatases↗
PubMed1977

The use of a phospholipase A-less Escherichia coli mutant to establish the action of granulocyte phospholipase A on bacterial phospholipids during killing by a highly purified granulocyte fraction.

Phospholipase A2 present in a highly purified, potently bactericidal, fraction from rabbit graulocytes produces net bacterial phospholipid degradation during killing of a phospholipase A-less strain of Escherichia coli. In the wild-type parent strain phospholipid breakdown is caused not only by the action of phospholipase A2 but also by phospholipase A1, indicating activation of the most prominent phospholipase of E. coli. This activation occurs as soon as the bacteria are exposed to the granulocyte fraction. Phospholipid breakdown by both phospholipases A is dose dependent but reaches a plateau after 30-60 min and at higher concentrations of the fraction. Phospholipid degradation is accompanied in both strains by an increase in permeability to actinomycin D that is also dose dependent. Even though net hydrolysis of phospholipids is greater in the parent strain than in the mutant, the increase in permeability is the same in the two strains. The addition of 0.04 M Mg2+, after the effects on phospholipids and permeability have become manifest, initiates in both strains the restoration of insensitivity to actinomycin D, the net resynthesis of phospholipids, and the disappearance of monoacylphosphatides and the partial disappearance of free fatty acids that had accumulated. Loss of ability to multiply is not reversed by Mg2+ in either strain. Less than 5 micrograms of granulocyte fraction causes loss of viability of from 90 to 99% of 1 X 10(8) microorganisms of both strains. However, at lower concentrations the parent strain is considerably more sensitive to the bactericidal effect of the granulocyte fraction than the mutant strain.

Animals↗
PubMed1979

Negatively charged phospholipids and their position in the cholesterol affinity sequence.

The effects of low concentrations of cholesterol in mixtures of a negatively charged phospholipid (phosphatidylserine or phosphatidylglycerol) and another phospholipid (phosphatidylcholine, sphingomyelin or phosphatidylethanolamine) have been studied by differential scanning calorimetry. Only mixtures which showed a gel phase miscibility gap have been employed. It was demonstrated that in mixtures with phosphatidylethanolamine, cholesterol was preferentially associated with the negatively charged phospholipid, regardless whether this species represented the component with the high or with the low transition temperature in the mixture. In mixtures of a negatively charged phospholipid and phosphatidylcholine, cholesterol associated with the negatively charged phospholipid; when the phosphatidylcholine was the species with the low transition temperature, cholesterol had an affinity for the phosphatidylcholine and for the negatively charged phospholipid as well. Cholesterol, in a mixture of sphingomyeline with a high and phosphatidylserine with a low transition temperature, was preferentially associated with sphingomyelin. From these experiments it is concluded that phospholipids show a decrease in affinity for cholesterol in the following order: sphingomyelin greater than phosphatidylserine, phosphatidylglycerol greater than phosphatidylcholine greater than phosphatidylethanolamine.

Calorimetry, Differential Scanning↗