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PubMed1996

Fatty acid double bond orientation alters interaction with L-cell fibroblasts.

Relatively little is known of fatty acid specificity in cellular fatty acid uptake. In this study L-cells, a fibroblastic cell line with very low levels of endogenous cytosolic fatty acid binding protein, were used to examine the role of cis and trans unsaturation on fatty acid uptake. The fluorescent fatty acids, trans-parinaric acid and cis-parinaric acid, were used as analogs of straight-chain saturated, and kinked-chain unsaturated fatty acids, respectively, in order to evaluate the fatty acid specificity of the uptake system. Parinaric acid is poorly metabolizable; greater than 97% was unesterified while 3H-oleic acid was almost totally metabolized after 30 min uptake. Cis- and trans-parinaric acid uptake was saturable and dependent on the concentration of fatty acid. However, the initial rate and maximal amount of trans-parinaric acid taken up by the L-cells was greater than for cis-parinaric acid under the same conditions. The affinity of L-cell uptake for trans-parinaric acid (Km = 0.12 uM) was 35-fold higher than that for cis-parinaric acid (Km = 4.17 uM). Based on competition studies with oleic and stearic acids, it was concluded that the cis- and trans-parinaric acid were taken up by the same L-cell fatty acid uptake system. The results suggest that the L-cell fatty acid uptake system has selectivity for straight chain rather than kinked chain unsaturated fatty acids.

Animals↗
PubMed2001

Inverse association between trans isomeric and long-chain polyunsaturated fatty acids in cord blood lipids of full-term infants.

BACKGROUND: Previous studies showed significant inverse correlations between values of trans isomeric and long-chain polyunsaturated fatty acids in plasma lipids of preterm infants and healthy children aged 1-15 y. OBJECTIVE: We sought to evaluate the same correlations in full-term infants at birth. DESIGN: We studied healthy full-term infants (n = 42) born after normal pregnancies and deliveries. All infants had a family history of atopy (both parents or one of the parents and a sibling had atopic symptoms). The fatty acid composition of venous cord blood lipids was determined by high-resolution capillary gas-liquid chromatography. RESULTS: The mean (+/-SEM) sum of trans fatty acids was 0.49 +/- 0.02% by wt in phospholipids, 2.47 +/- 0.20% by wt in cholesterol esters, 1.73 +/- 0.09% by wt in triacylglycerols, and 1.59 +/- 0.07% by wt in nonesterified fatty acids. Linear correlation analysis showed significant inverse correlations between the sum of trans fatty acids and both arachidonic acid and docosahexaenoic acid in phospholipids (r = -0.56, P < 0.001, and r = -0.48, P = 0.01, respectively), cholesterol esters (r = -0.52, P < 0.001, and r = -0.39, P = 0.018, respectively), and nonesterified fatty acids (r = -0.41, P = 0.007, and r = -0.41, P = 0.006, respectively). CONCLUSION: Because trans fatty acids in the fetal circulation must originate from the maternal diet, our results indicate that maternal exposure to trans fatty acids may represent a previously neglected variable that inversely influences long-chain polyunsaturated fatty acid status in full-term infants at birth.

Cholesterol Esters↗
PubMed1981

The influence of trans-acids on desaturation and elongation of fatty acids in developing brain.

trans-Monounsaturated acids account for up to 3% of the total octadecenoic acyl chains of human brain lipids. To investigate the influence of trans-acids on desaturation and chain elongation of fatty acids, in vitro and in vivo experiments with rat brain were performed. In the in vitro assays of delta 9 desaturation, delta 6 desaturation and chain elongation, trans,trans-dienoic acid was inhibitory, particularly to chain elongation. Slight differences between the inhibitory effects of trans-monoenoic acids and their cis-isomers were observed. In an in vivo model, unlabeled fatty acid (stearate, oleate, elaidate, linoleate, linoelaidate, arachidonate, or trans-monoene from margarine) was injected simultaneously with [1-14C] linoleic acid into the brains of suckling rats. Linoelaidate and oleate inhibited desaturation and elongation of linoleate, whereas elaidate, stearate and trans-monoene from margarine were stimulatory. While the demonstration of differences between cis and trans monenic isomers required relatively high levels of the test acids, it appears that trans-acids can influence desaturation and elongation enzymes that lead to acyl chain modification in the central nervous system.

Animals↗
PubMed1984

Effects of free fatty acids on release of superoxide and on change of shape by human neutrophils. Reversibility by albumin.

Release of superoxide from human neutrophils was stimulated by cis-unsaturated fatty acids (e.g. arachidonate, linoleate). All the saturated and trans-unsaturated fatty acids tested were ineffective in this context. Binding of linoleate to neutrophils was biphasic and could be resolved into a linear and a saturable component. The extent of linoleate binding to the saturable component correlated strongly with the amount of O-2 released (r = 0.96). Palmitate, a saturated fatty acid, exhibited only linear binding to neutrophils and the binding was similar to the linear component of linoleate binding. All cis-unsaturated fatty acids tested decreased the fluorescence polarization of cis- and trans-parinaric acid used as membrane probes in suspensions of neutrophils, whereas the saturated fatty acid, myristate, increased the polarization. Fatty acids which stimulated O-2 release induced morphological changes (i.e. evaginations) in neutrophils, whereas the inactive fatty acids did not affect the cellular morphology. Effects on both the cell morphology and superoxide release were reversed by treatment of the cells with delipidated albumin. These data are discussed in relation to recent reports concerning the effects of free fatty acids on various biochemical systems.

Arachidonic Acid↗
PubMed1982

Metabolism of very long-chain monounsaturated fatty acids (22:1) and the adaptation to their presence in the diet.

Unadapted rats and other animal species have a limited capacity to metabolize monounsaturated fatty acids with 22 carbons (22:1). Excess amounts in the diet of fats containing these fatty acids cause a transient accumulation (lipidosis) of triacylglycerol in the heart and other tissues but not in the liver, which seems to export the 22:1 fatty acids as very low density lipoproteins to the blood plasma. The acute lipidosis most probably is explained by a slow oxidation of 22:1 acyl-CoA by the mitochondrial acyl-CoA dehydrogenase combined with an inhibitory effect of this CoA ester on the oxidation of acyl-CoA esters of a more "normal" chain length. Other fatty acid metabolizing enzymes also show slow reaction rates with the 22:1 fatty acids. Upon continued feeding of diets with 22:1 fatty acids, an adaptation takes place and the lipidosis disappears. This adaptation coincides with the development of an increased capacity to chain-shorten the 22:1 fatty acids, especially in the liver, but also in the heart. The chain-shortening seems to be due to a partial beta-oxidation of the 22:1 fatty acids by the peroxisomal beta-oxidation enzyme system which shows an increased activity in adapted rats. In such rats, less 22:1 fatty acids circulate in the plasma very low density lipoproteins than in unadapted rats. The drug clofibrate (ethyl-p-chlorophenoxyisobutyrate) which induces increased activity of the peroxisomal beta-oxidation enzymes, provides partial protection against the lipidosis in unadapted animals. Hydrogenated fish oil (containing different 22:1 isomers and many fatty acids with trans double bonds) is more efficient as an inducer of the chain-shortening of erucic acid in the liver than is rapeseed oil, which contains only one 22:1 fatty acid isomer and no fatty acids with trans double bonds. The hydrogenated fish oil causes less lipidosis than does rapeseed oil when diets containing the same amount of 22:1 fatty acids are fed. It is suggested that CoA esters that are poorly oxidized by the mitochondria (e.g., esters of erucic acid, of some fatty acids with trans double bonds, and of clofibric acid) may trigger the adaptation process.-Bremer, J., and K. R. Norum. Metabolism of very long-chain monounsaturated fatty acids (22:1) and the adaptation to their presence in the diet.

Acyl Coenzyme A↗
PubMedDate not supplied

Effect of nutritional factors on biochemical, structural and metabolic characteristics of muscles in ruminants, consequences on dietetic value and sensorial qualities of meat.

Ruminant meat is an important source of nutrients and is also of high sensory value. However, the importance and nature of these characteristics depend on ruminant nutrition. The first part of this review is focused on biochemical and dietetic value of this meat. It offers a panel of quantitative and qualitative contributions, especially through its fatty acids characteristics. Since saturated and trans-monounsaturated fatty acids are considered as harmful to human health, their amount in muscles can be reduced by increasing the proportions of dietary polyunsaturated fatty acids (PUFA) absorbed by the animals. On the contrary, some fatty acids (n-6 and n-3 PUFA, conjugated linoleic acid) specifically incorporated in muscle tissues would play a favourable role in the prevention or reduction of major diseases in human (cancers, atherosclerosis, obesity) and therefore be recommended. The second part of this review treats different aspects of the sensorial qualities of meat. Skeletal muscle structure and its biochemical components influence muscle transformation to meat and sensorial qualities including tenderness, colour, flavour and juiciness. This paper shows how nutrition can influence, in ruminants, metabolic activity as well as muscle structure and composition, and thereby affect meat quality.

Animal Nutritional Physiological Phenomena↗
PubMed2001

Pseudomonas alcaliphila sp. nov., a novel facultatively psychrophilic alkaliphile isolated from seawater.

Facultatively psychrophilic alkaliphilic strains were isolated from seawater obtained off the coast of Rumoi, Hokkaido, Japan. They were Gram-negative, aerobic straight rods with polar flagella. The isolates were catalase- and oxidase-positive and able to grow at 4 degrees C, but not at 40 degrees C. They produced acid from D-glucose under aerobic conditions. The isolates reduced nitrate to nitrite and hydrolysed casein and gelatin, but not starch or DNA. NaCl was required for growth at pH 10 but was not required at neutral pH. The major isoprenoid quinone was ubiquinone-9 (Q-9) and the DNA G+C content was 62.3-63.2 mol%. The whole-cell fatty acids mainly consisted of C16:0, C16:1(9c) and C18:1(9c), with 3-OH C10:0 and 3-OH C12:0 as the hydroxyl fatty acids. A larger amount of trans-unsaturated fatty acid, C16:1(9t) was observed when the cells were grown at pH 7 compared to when cells were grown at pH 10. Phylogenetic analysis based on 16S rRNA gene sequencing indicated that the bacteria are members of the genus Pseudomonas. Analysis of DNA-DNA relatedness data with several close phylogenetic neighbours revealed a low level of hybridization (less than 61%). On the basis of phenotypic characteristics, phylogenetic analysis and DNA-DNA relatedness data, it is concluded that these isolates represent a separate new species. Accordingly, the name Pseudomonas alcaliphila is proposed. The type strain is AL15-21T (= JCM 10630T = IAM 14884T).

Alkalies↗
PubMed1980

cis-Unsaturated fatty acids inhibit cap formation on lymphocytes by depleting cellular ATP.

The inhibition of anti-immunoglobulin cap formation on lymphocytes by cis-unsaturated fatty acids, but not saturated or trans-unsaturated fatty acids, which recently been reported (Klausner, R. N., Bhalla, D. K., Dragsten, P., Hoover, R. L., and Karnovsky, M. J. (1980) Proc. Natl. Acad. Sci. U. S. A. 77, 437-441), can be accounted for quantitatively by the effects of the fatty acids on ATP levels in the intact cells. Only the cis-unsaturated fatty acids lowered the cellular ATP level sufficiently (to < 80%) to inhibit cap formation significantly. The profile for the dependence of cap formation on cellular ATP level obtained with the fatty acids is very similar to the corresponding profile described previously for several capping ligands and a range of metabolic inhibitors used to depress the cellular ATP levels. Oxygen electrode experiments indicate that the unsaturated, but not the saturated, fatty acids can uncouple lymphocyte mitochondria in intact cells, whereas both types of fatty acids can uncouple isolated rat liver mitochondria. The difference in the effect of the two types of fatty acids on ATP levels in the intact cells is attributed to the inability of the saturated fatty acids to penetrate in sufficient concentrations to the mitochondria to cause uncoupling. The protective action of external calcium against inhibition of capping by the free fatty acids is attributed to the effect of calcium in cross-bridging the anionic fatty acids and reducing their effective concentration. These interpretations account for all of the experimental data and are much simpler than the model proposed by Klausner et al.

Adenosine Triphosphate↗
PubMed1987

Inhibition of platelet aggregation by unsaturated fatty acids through interference with a thromboxane-mediated process.

cis- and trans-unsaturated fatty acids with 18 carbon atoms (oleic, linoleic, elaidic and linolelaidic acid) inhibited aggregation of washed rabbit platelets stimulated with collagen, arachidonic acid and U46619 when in the same concentration ranges. Thrombin-induced aggregation was not affected by any of them. Saturated fatty acid (stearic acid) had no effect on this response. The inhibition is independent of the induced change in membrane fluidity, since trans-isomers could not induce the change in fluorescence polarization of 1,6-diphenyl-1,3,5-hexatriene. Unsaturated fatty acids, except linoleic acid, did not interfere with the formation of thromboxane B2 from exogenously added arachidonic acid. All the unsaturated fatty acids only slightly inhibited the arachidonic acid liberation by phospholipase A2 in platelet lysate. This indicates that the unsaturated fatty acids may block a process after formation of thromboxane A2 in response to collagen and arachidonic acid. The increase in phosphatidic acid formation stimulated with U46619 was inhibited dose dependently by each of the unsaturated fatty acids but that stimulated with thrombin was not affected by any of them. Phospholipase C activity measured by diacylglycerol formation in unstimulated platelet lysate was not inhibited by the fatty acids. The elevation of cytosolic free Ca2+ induced by arachidonic acid or U46619 and Ca2+ influx by collagen were inhibited almost completely at the same concentration as that which inhibited their aggregation. These data suggest that the unsaturated fatty acids were intercalated into the membrane and inhibited collagen- and arachidonic acid-induced platelet aggregation by causing a significant suppression of the thromboxane A2-mediated increase in cytosolic free Ca2+, probably due to interference with the receptor-operated Ca2+ channel.

Animals↗
PubMed2006

Acute postprandial effect of hydrogenated fish oil, palm oil and lard on plasma cholesterol, triacylglycerol and non-esterified fatty acid metabolism in normocholesterolaemic males.

The majority of research has focused on the association between trans unsaturated fatty acids (TUFA) from hydrogenated vegetable oils and heart disease even though TUFA are also produced from hydrogenated fish oil. We compared the acute effect of three solid fats on postprandial cholesterol, triacylglycerol (TAG) and NEFA concentrations in normocholesterolaemic males. Eight healthy male volunteers consumed each of the three 40 g fat meals (partially hydrogenated fish oil (PHFO), palm oil and lard) in random order and blood samples were drawn at 2, 4, 6 and 8 h thereafter for lipid analysis. The postprandial response in plasma TAG, TAG-rich lipoprotein-TAG (TRL-TAG), total cholesterol and plasma NEFA, measured as the area under the postprandial curve, was not significantly different between the three meals (P>0.05), which varied in MUFA, PUFA and TUFA content. There was no marked elevation of longer-chain fatty acids (C20-22, cis or trans isomers) into the TRL-TAG fraction following the PHFO meal even though they provided 40 % of the total fatty acids in the PHFO meal. The postprandial TRL-TAG response to PHFO was expected to be higher, as it is higher in TUFA, lower in PUFA and similar in saturated fatty acid composition compared with the lard and palm oil test meals. The absence of a higher postprandial response following ingestion of PHFO could be as a result of reduced absorption and increased oxidation of long-chain fatty acids (both cis and trans isomers).

Adult↗
PubMed1995

trans isomers of oleic and linoleic acids in adipose tissue and sudden cardiac death.

trans isomers of unsaturated fatty acids are formed by biological or industrial hydrogenation. A population case-control study of sudden cardiac death in mean was done to test the hypothesis that trans isomers of oleic acid and linoleic acid increase the risk of sudden cardiac death due to coronary artery disease. In adipose tissue obtained at necropsy from 66 cases of sudden cardiac death and taken from 286 healthy age and sex matched controls, the proportions of trans isomers of oleic and linoleic acid were measured by gas-liquid chromatography. In cases, the mean (SE) percentage of total trans fatty acids (C18:1 plus C18:2), expressed as a proportion of all fatty acids, was significantly lower (2.68 [0.08]%) than in healthy controls (2.86 [0.04]%; p < 0.05). trans C18:1 was 2.1 (0.7)% in cases compared with 2.27 (0.04)% (p < 0.05) in controls. The proportion of all trans isomers of linoleic acid was 0.58 (0.02)% in cases compared with 0.59 (0.01)% in controls (p = 0.98). The estimated relative risk for sudden cardiac death of trans C18:1 and C18:2 fatty acids combined did not differ significantly from 1.0 in relation to the distribution of these trans isomers by quintile in the control population. The relative risk (95% CI) of sudden cardiac death in the top quintile was 0.40 (0.15-1.02) for C18:1 and 1.08 (0.48-2.74) for C18:2 compared with the bottom quintiles of their respective control distributions. When these univariate relations for trans fatty acids were adjusted for coronary risk factors, smoking was the only factor that remained independently associated with risk of sudden cardiac death (2.27 [1.23-4.17]). Overall, there was no evidence of a relation between trans isomers of oleic and linoleic acids combined and sudden cardiac death. However, trans oleic acid was negatively associated with risk of sudden cardiac death, whereas no association with trans forms of linoleic acid was seen. This study does not support the hypothesis that trans isomers increase the risk of sudden cardiac death.

Adipose Tissue↗
PubMed1984

Influence of dietary fats on ultrastructure and fatty acid. Composition of swine arterial tissue.

The effects of trans-fatty acid-rich and saturated acid-rich diets on the fatty acid composition and morphology of swine arterial tissues were studied. Three groups of two-month-old swine were fed either a basal diet or a basal diet containing 8.3% margarine or butter for 4 months. The most significant change observed in the fatty acid pattern was the accumulation of 18 : 2 omega 6 and the suppression of both 20 : 4 omega 6 and 22 : 4 omega 6 acids in the aortic tissues of swine fed either butter or margarine as compared to swine fed the basal diet. Also, omega 6 metabolites were significantly decreased in swine fed diets containing butter or margarine, as compared to those fed a basal diet. The group of swine which were fed either butter or margarine containing diets suffered a significant increase in intimal thickening of the coronary artery compared to those fed the basal diet. These thickened intima were characterized by the presence of modified smooth muscle cells, lipid containing cells, and degenerated cells without stainable lipids. However, there was no significant difference in the incidence and extent of intimal thickening of the coronary arteries between swine fed either of the two fat containing diets.

Animals↗
PubMed1986

Effect of fatty acid supplementation on thermotropic behavior of membrane lipids and leucine transport in Saccharomyces cerevisiae.

An unsaturated fatty acid-requiring mutant (KD 115) of Saccharomyces cerevisiae shows altered phospholipid composition, transport behavior, and physical properties of membrane lipids when grown in the presence of different cis- and trans-unsaturated fatty acids. There is an increase in phosphatidyl ethanolamine content and a concomitant decrease in phosphatidyl choline content in the cells supplemented with trans-unsaturated fatty acids. The affinity for uptake of L-leucine is higher in the cis-unsaturated fatty acid-supplemented cells compared with the trans-unsaturated fatty acid-supplemented cells. The temperature-dependence of L-leucine uptake bears a reasonably good correlation with the thermotropic behavior of the membrane lipids as studied by the steady-state fluorescence polarization technique. The present findings are discussed in light of the importance of the lipid environment in modulating membrane-associated functions.

Biological Transport↗
PubMed2005

Changes in conjugated linoleic acid and palmitoleic acid are correlated to retinol levels in chronic renal failure in both hemodialysis and conservative treatment patients.

An increase in conjugated linoleic acid (CLA), a natural fatty acid present in our diet, which possesses anticarcinogenic and antiatherogenic activities in experimental models, has been found in both the plasma and adipose tissue of end-stage chronic renal failure (ESCRF) patients. Increased levels of retinol have also been found in those patients, due to a reduced excretion of the retinol-binding protein. Since retinol is known to influence lipid metabolism, we evaluated whether changes in retinol, CLA, and other fatty acids are correlated in the plasma of CRF patients. We measured CLA, retinol, and unsaturated fatty acids in the plasma of the following groups: (A) 35 ESCRF patients; (B) 20 hemodialysis (HD) patients; (C) 20 healthy controls. Subjects with total cholesterol and/or triglycerides higher than 250 mg/dL were excluded. We found a significant increase in CLA, retinol, palmitoleic (16:1), and oleic (18:1) acids in ESCRF patients. In HD patients we found a similar pattern, however, CLA increase was not significant. No changes were observed in the other fatty acids measured. In the groups of ESCRF and HD patients, a positive correlation between the levels of plasma retinol and CLA, and between retinol and 16:1 was found. These correlations were not detected in controls. The abnormal levels of plasma retinol in CRF patients might partly explain the changes in CLA and 16:1. The influence of retinol levels on these fatty acids might be due to an induction of delta 9 desaturase. In fact, 16:1 is known to be produced, partly, by delta 9 desaturation of palmitic acid. Moreover, the formation of CLA from delta 9 desaturation of vaccenic acid-a trans-monounsaturated fatty acid present in our diet-has recently been demonstrated in humans. Nevertheless, our data do not represent direct evidence supporting an increased delta 9 desaturase activity in CRF patients. Another possible explanation might be a variation in the exogenous intake.

Adipose Tissue↗
PubMed1997

cis-trans isomerization of unsaturated fatty acids: cloning and sequencing of the cti gene from Pseudomonas putida P8.

Transposon mutants of Pseudomonas putida P8 were generated by applying a mini-Tn5 mutagenesis system. The mutants obtained were checked for their ability to tolerate increased temperatures and elevated phenol concentrations. Approximately 5,800 transposon mutants were used to generate a pool of 600 temperature-sensitive strains; one of these strains was identified as being damaged in its ability to perform cis-trans isomerization of fatty acids. A gene library of P. putida P8 was constructed and screened by using as a probe sequences immediately adjacent to the mini-Tn5 insertion. A DNA fragment that complemented the mutation was isolated and cloned. The corresponding gene, termed cti, is located close to the methionine synthase locus (metH) in P. putida P8. A cti-carrying fragment integrated into a plasmid also conferred the ability for cis-trans isomerization to Escherichia coli; the cti gene was completely sequenced, and the amino acid sequence was deduced.

Amino Acid Sequence↗
PubMed1998

Assignment of Vibrio sp. strain ABE-1 to Colwellia maris sp. nov., a new psychrophilic bacterium.

A psychrophilic bacterium, previously described as Vibrio sp. strain ABE-1T, has been reassigned by phenotypic characterization, chemotaxonomic analysis and 16S rRNA phylogenetic analysis. The organism was curved rods and it could reduce nitrate to nitrite and hydrolyse gelatin and DNA, but not chitin. NaCl was required for growth. This strain was susceptible to the vibriostatic compound O/129. The major isoprenoid quinone was ubiquinone-8 and the DNA G + C content was 39.4 mol%. The whole-cell fatty acids comprised saturated and monounsaturated fatty acids with 10-18 C atoms; saturated and monounsaturated C16 fatty acids were predominant. Strain ABE-1T contained the unique trans-unsaturated fatty acid, 9-trans-hexadecenoic acid. Although strain ABE-1T has been identified as a Vibrio species, the strain did not ferment glucose. Phylogenetic analysis based on 16S rRNA sequencing indicated that strain ABE-1T was more closely related to Colwellia species than to Vibrio species. However, strain ABE-1T differed from other reported Colwellia species in terms of phylogenetic position, some phenotypic characteristics, chemotaxonomic analysis and relatedness by DNA-DNA hybridization. Accordingly, the name Colwellia maris is proposed. The type strain is ABE-1T (= JCM 10085T).

Base Composition↗
PubMed1987

Activities of liver mitochondrial and peroxisomal fatty acid oxidation enzymes in rats fed trans fat.

The effect of trans fat on the activities of liver mitochondrial and peroxisomal fatty acid oxidation enzymes was examined in various strains of rats. When Wistar and Sprague-Dawley rats were fed for 30 days diets containing either olive oil or partially hydrogenated corn oil as a source of cis- or trans-octadecenoate, respectively, the activities of various enzymes of mitochondrial and peroxisomal beta-oxidation measured with cis- and trans-9-octadecenoic acid as substrates showed little dietary fat-dependent change. In Fischer 344 rats, feeding trans fat for 15 mo increased only moderately various enzymes of beta-oxidation except for carnitine acyltransferase. The rate of mitochondrial ketogenesis and the activity of carnitine acyltransferase measured with trans-9-octadecenoic acid as a substrate were about half those with the cis-counterpart. Peroxisomes oxidized trans-9-octadecenoyl-CoA at a rate comparable to the cis-counterpart. It was concluded from this study and previous ones that the difference in the geometry of dietary fatty acid had only a marginal effect in modulating the hepatic fatty acid oxidation system, in spite of marked differences in the metabolic behavior of cis- and trans fatty acid in cell-free preparations and perfused liver.

Animals↗
PubMed1983

Calcium modulates fatty acid dynamics in rat liver plasma membranes.

Modulation of free fatty acid binding in isolated rat liver plasma membranes was evaluated using the fluorescent fatty acids trans-parinaric and cis-parinaric acid as analogues for saturated and unsaturated fatty acids, respectively. Binding of trans-parinarate but not cis-parinarate was inhibited by physiological levels of Ca2+. The effect was reversed by addition of excess EGTA. Calcium decreased the aqueous to lipid partition coefficient, Kp, of trans-parinaric acid for liver plasma membranes while increasing the Kp for cis-parinaric acid. In addition, Ca2+ also altered the fluorescence lifetime, the quantum yield, and the relative partitioning of trans-parinaric and cis-parinaric acid into fluid and solid phases. Calcium and EGTA did not affect the binding of 1,6-diphenyl-1,3,5-hexatriene. The effect of Ca2+ on the liver plasma membrane structure was to increase the rigidity of the membrane, primarily the solid domain. The fluorescence polarization of trans-parinarate, cis-parinarate, and 1,6-diphenyl-1,3,5-hexatriene at 24 degrees C in liver plasma membranes in the absence of Ca2+ was 0.295 +/- 0.008, 0.253 +/- 0.007, and 0.284 +/- 0.005, respectively. Calcium (2.4 mM) increased the polarization of these probe molecules in liver plasma membranes by 8-10%. EGTA (3.4 mM) reversed or abolished the increase in polarization. Thus, the fluorescent fatty acids trans-parinarate and cis-parinarate may be used to monitor fatty acid binding by isolated membranes, to evaluate factors such as Ca2+ which modulate fatty acid binding, and to investigate the microenvironment in which the fatty acids residue. The data suggest that Ca2+ may be an important regulator of fatty acid uptake by the liver plasma membrane, and thereby interact with intermediary metabolism of lipids at a step not involving lipolytic or synthetic enzymes.

Animals↗