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Lipid-protein interactions in membranes: effect of lipid composition on mobility of spin-labeled cysteine residues in yeast plasma membrane.

In order to gain direct evidence for lipid-dependent protein conformation in membrane, effects of modification of lipid composition on mobility of spin-labeled cysteine residues were investigated in the plasma membrane of the yeast Saccharomyces cerevisiae. Conversion of the bulk of phospholipids to diglycerides by treatment of the membrane with phospholipase C substantially enhanced spectral anisotropy. However, alterations of the viscosity of the lipid-bilayer by enriching the membrane with palmitelaidic or oleic acid had no effect on mobility of spin-labeled cysteine residues. These observations indicate that while the spin-labeled residues are not in direct contact with the lipid core of the membrane, there are lipid-protein interactions to the extent that removal of the polar portion of the bulk of phospholipids induces conformational changes in proteins, which in turn restrict mobility of these residues. It is concluded that conformation of membrane proteins on lipid structure and that phospholipids have a role in preserving the native conformation of proteins.

Adenosine Triphosphatases↗

Reduction of plasma cholesterol by Curcuma comosa extract in hypercholesterolaemic hamsters.

The influence of the extract of Curcuma comosa Roxb. (Zingiberaceae) on lipid metabolism was investigated in hypercholesterolaemic hamsters. Intragastric administration of the ethyl acetate extract of C. comosa rhizome (0-500 mg/kg per day) to hypercholesterolaemic animals for 7 days decreased both plasma triglyceride and cholesterol levels in a dose-dependent manner. The reduction of plasma cholesterol levels was accompanied by a significant increase in the hepatic cholesterol content while the triglyceride content was not significantly changed. The increase of the hepatic cholesterol content was brought about by an expansion of the free cholesterol pool which specifically augments biliary cholesterol excretion. The C. comosa extract also increased plasma high density lipoprotein (HDL)-cholesterol and decreased plasma low density lipoprotein (LDL)-cholesterol. These results suggest that the C. comosa extract exerts a hypolipidaemic action by acceleration of lipid mobilization from extrahepatic tissue to the liver which subsequently increases excretion of cholesterol via the bile for excretion.

Animals↗

Subnanometer actuation of a tethered lipid bilayer monitored with fluorescence resonance energy transfer.

Lipid membrane nanotechnology can play a key role in preserving the function of transmembrane proteins on biofunctional substrates. We show here that rational nanoscopic actuation of a polymer-tethered lipid bilayer can be achieved by modulating the dielectric environment at the membrane-substrate interface. This provides a hydrated platform with increased lipid mobility compared to bilayers supported directly onto silica. We suggest that this construct may be used for promoting the functional reconstitution of transmembrane proteins on planar surfaces for bioanalytical devices.

Avidin↗

A novel adipokinetic octapeptide found in the damselflies Pseudagrion inconspicuum and Ischnura senegalensis.

A member of the adipokinetic hormone family of peptide was identified in the damselflies Pseudagrion inconspicuum and Ischnura senegalensis using a heterologous (in migratory locusts and American cockroaches) and a homologous (in P. inconspicuum) bioassay. After isolation of the peptide by reversed-phase h.p.i.c. of corpora cardiaca, its structure was determined by automated Edman degradation and matrix-assisted laser-desorption ionization m.s. The sequence of a blocked uncharged octapeptide was established: pGlu-Val-Asn-Phe-Thr-Pro-Gly-TrpNH2. One corpus cardiacum of P. inconspicuum contains about 2.4 pmol of this novel peptide. The synthetic peptide was chromatographically indistinguishable from the natural compound, and on injection in low quantities into the damselfly P. inconspicuum, haemolymph levels of lipids were increased. It is concluded that the novel peptide may be involved in controlling lipid mobilization during flight. Hence the peptide is designated Psi-AKH, Pseudagrion inconspicuum adipokinetic hormone.

Amino Acid Sequence↗

Lipolytic response of adipocytes to epinephrine in sedentary and exercise-trained subjects: sex-related differences.

Adipose tissue lipolytic activity is increased in endurance-trained subjects, but little is known about the mechanisms of this increase. To understand more fully the mechanisms involved and to discover whether sex-related differences exist, biopsies of fat were performed in the periumbilical region of 20 sedentary subjects (10 women (W) and 10 men (M)) and 20 trained subjects (10 W, 10 M); the in vitro response to epinephrine of the collagenase-isolated fat cells was studied. Glycerol release, chosen as an adipocyte lipolysis indicator, was measured by bioluminescence. Dose-response curves with epinephrine (alpha 2 and beta agonist), with isoproterenol (beta agonist) and epinephrine + propranolol and adenosine deaminase, were studied. Epinephrine-induced lipolysis was enhanced in trained subjects and this was due to an increased efficiency of the beta-adrenergic pathway. However, differences were found between the two sexes. In trained men, the lipolysis increase resulted from the enhancement of the beta-adrenergic pathway efficiency without any significant decrease in the alpha 2-adrenergic pathway efficiency. In trained women, the lipolysis increase was not only due to the enhancement of the beta-adrenergic pathway efficiency (which was greater than in trained men), but also to a significant decrease in the alpha 2-adrenergic pathway efficiency. Despite the decrease, the alpha 2-adrenergic pathway remained more efficient in trained women than in trained men, as was the case in sedentary subjects. It is concluded that endurance training led to better lipid mobilization and that this effect seemed greater in women than in men.

Adenosine Deaminase↗

The effects of high fat feeding during one cycle of reproduction consisting of pregnancy, lactation and recovery on body composition and fat pad cellularity in the rat.

The changes in adipose depot weight, cell size, cell number and body composition during pregnancy, lactation and recovery were studied in Osborne-Mendel rats fed standard or high fat diets. Rats were killed on day 21 of pregnancy, after 21 days of lactation, and after 21 or 22 days of a postlactational recovery period. Nonpregnant control groups were killed at the beginning and at the conclusion of the experimental period. The high fat-fed, mated group was always fatter than similarly treated animals fed standard diets throughout pregnancy and lactation. However, by the end of the recovery period, carcass composition of the animals fed high fat or standard diets and the nonpregnant groups were not statistically different. The weight of the parametrial, retroperitoneal and subscapular depots was higher in the high fat-fed animals at the end of the recovery period, and in the latter two pads, this increase was statistically significant. Thus, despite the extensive lipid mobilization that occurs during lactation, the high fat-fed animals appear to be predisposed to postpartum obesity.

Adipose Tissue↗

Isolation and primary structure of a peptide from the corpora cardiaca of Heliothis zea with adipokinetic activity.

An adipokinetic hormone was isolated from the corpora cardiaca of the corn ear worm moth, Heliothis zea, and purified by reversed phase high performance liquid chromatography. The primary structure, pGlu-Leu-Thr-Phe-Thr-Ser-Ser-Trp-Gly-NH2, was determined by automated gas-phase Edman degradation of the peptide deblocked with pyroglutamic aminopeptidase, and by fast atom bombardment mass spectrometry. The hormone was synthesized and the natural and synthetic material had identical chromatographic, spectroscopic, and biological properties. The peptide was found to have lipid mobilizing activity in H. zea adults.

Amino Acid Sequence↗

Lipid diffusion in giant unilamellar vesicles is more than 2 times faster than in supported phospholipid bilayers under identical conditions.

The lateral diffusion coefficients of a BODIPY tail-labeled lipid in two model systems, namely, free-standing giant unilamellar vesicles (GUVs) and supported phospholipid bilayers (SPBs), were determined by fluorescence correlation spectroscopy (FCS) using the Z-scan approach. For the first time, the performed measurements on 1,2-dioleoyl-sn-glycero-3-phosphocholine (DOPC) bilayers maintain exactly the same experimental conditions for both systems, which allows for a quantitative comparison of lipid diffusion in these two commonly used model membranes. The results obtained revealed that the lipid mobility in free-standing bilayers (D=7.8+/-0.8 microm2 s-1) is significantly higher than in the bilayer created on the solid support (mica) (D=3.1+/-0.3 microm2 s-1).

Diffusion↗

Effects of capsaicin on lipid metabolism in rats fed a high fat diet.

Effects of capsaicin, a pungent principle of hot red pepper, were studied in experiments using male rats fed a diet containing 30% lard. Capsaicin was supplemented at 0.014% of the diet. The level of serum triglyceride was lower when capsaicin was present in the diet than when it was not. Levels of serum cholesterol and pre-beta-lipoprotein were not affected by the supplementation of capsaicin. The perirenal adipose tissue weight was lower when capsaicin was present in the diet than when it was not. Hepatic enzyme activities of glucose-6-phosphate dehydrogenase and adipose lipoprotein lipase were lower in rats fed the 30% lard diet than in those fed a nonpurified diet. Activities of these two enzymes were higher when capsaicin was added to the diet than when it was not. Hepatic acetyl-CoA carboxylase, beta-hydroxyacyl-CoA dehydrogenase, and adipose hormone-sensitive lipase activities were not affected by capsaicin feeding. Lipid absorption was not affected by the supplementation of capsaicin. The perirenal adipose tissue weight and serum triglyceride were decreased as the level of capsaicin in the diet increased up to 0.021%. These results suggest that capsaicin stimulates lipid mobilization from adipose tissue and lowers the perirenal adipose tissue weight and serum triglyceride concentration in lard-fed rats.

3-Hydroxyacyl CoA Dehydrogenases↗

Evidence that amylin stimulates lipolysis in vivo: a possible mediator of induced insulin resistance.

The present study investigated the role of amylin in lipid metabolism and its possible implications for insulin resistance. In 5- to 7-h-fasted conscious rats, infusion of rat amylin (5 nmol/h for 4 h) elevated plasma glucose, lactate, and insulin (P <0.05 vs. control, repeated-measures ANOVA) with peak values occurring within 60 min. Despite the insulin rise, plasma nonesterified fatty acids (NEFA) and glycerol were also elevated (P < 0.001 vs. control), and these elevations (80% above basal) were sustained over the 4-h infusion period. Although unaltered in plasma, triglyceride content in liver was increased by 28% (P < 0.001) with a similar tendency in muscle (18%, P = 0.1). Infusion of the rat amylin antagonist amylin-(8-37) (125 nmol/h) induced opposite basal plasma changes to amylin, i.e., lowered plasma NEFA, glycerol, glucose, and insulin levels (all P < 0.05 vs. control); additionally, amylin-(8-37) blocked amylin-induced elevations of these parameters (P < 0.01). Treatment with acipimox (10 mg/kg), an anti-lipolytic agent, before or after amylin infusion blocked amylin's effects on plasma NEFA, glycerol, and insulin but not on glucose and lactate. We conclude that amylin could exert a lipolytic-like action in vivo that is blocked by and is opposite to effects of its antagonist amylin-(8-37). Further studies are warranted to examine the physiological implications of lipid mobilization for amylin-induced insulin resistance.

Amyloid↗

Nutrient intake and energy regulation in physical exercise.

Rates of energy expenditure as well as total daily energy cost can be considerable during periods of exercise. In trained athletes, expenditure can be as high as 380 kJ/min during short-term maximal exercise. Training programmes of several hours' duration lead to a daily nutrient intake of 25-35 MJ in most Olympic sports. The mobilization of the energetic fuels of the body is modulated by the nature of the exercise. ATP and creatine phosphate stores in muscle cells are depleted within seconds during maximal work. Glycogen is the main fuel for heavy exercise of a few minutes' duration where performance capacity is limited by the degree of lactate accumulation and intracellular acidosis. Oxidation of both glucose and free fatty acids supplies the energy needed for exercise lasting more than two minutes, the relative contribution of lipids increasing with a longer duration or a lower intensity of the muscular work. Intramuscular stores of glycogen and triglycerides may be almost completely depleted in long-lasting exercise, e.g. a 100 km run. Under these conditions, glycogen stores in the liver and triglycerides in adipose tissue contribute approximately 70% of the energy need whereas 5-10% of the supply comes from oxidation of amino acids. Although adequate nutrition for exercise could be achieved through the intake of a well-balanced diet, the regulation of energy utilization can be influenced by the sources of food energy, by dietary modifications before exercise or by nutrient supplements during exercise. Intake before exercise of fructose or medium-chain triglycerides, both only weakly insulinogenic compared to glucose, leads to changes in blood substrates and metabolites. However, neither glycogen depletion in the working muscles nor performance capacity was influenced by a single meal containing this particular carbohydrate or lipid. Mobilization of free fatty acids in adipose tissue can be enhanced by caffeine or depressed by nicotinic acid. Since the rate of free fatty acid oxidation in skeletal muscle depends on the blood concentration of this substrate, energy regulation during exercise and work output are considerably influenced by the ingestion of such substances.

Adipose Tissue↗

Release of potassium, lipids, and proteins from nonionic detergent treated chicken red blood cells.

The plasma membrane of erythrocytes, as of other cells, is thought to act as the barrier responsible for maintaining intracellular gradients of most ions and small molecular species between the cell and its environment. Controlled application of the nonionic detergent Brij 58 effectively opened the erythrocyte plasma membrane, as judged by electron microscopy and lipid mobilization, but the cytoplasm maintained much of its integrity for about 30 min. Release of K+ correlated well with release of protein into the surrounding medium. The results demonstrate that permeabilization of the erythrocyte plasma membrane does not result in an instantaneous equilibration of small ions, such as K+, between the cell and its environment. A comparison was made between erythrocytes treated with Brij 58 and Triton X-100. The lipid and protein solubilizing actions of Triton X-100 were not as easily separable in time as those of Brij 58. The results of treatment of the erythrocytes with different types of nonionic detergents suggest that the membranolytic and cytoplasmic protein destabilizing actions of nonionic detergents correspond with their hydrophilic-lipophilic balance numbers (HLB values).

Animals↗

Adipokinetic hormone controls lipid metabolism in adults and carbohydrate metabolism in larvae of Manduca sexta.

The peptide hormone which controls activation of fat body glycogen phosphorylase in starving larvae of Manduca sexta was isolated from larval corpora cardiaca and sequenced by FAB tandem mass spectrometry. It was found to be identical with Manduca AKH. This, together with earlier observations, demonstrates that in M. sexta AKH controls glycogen phosphorylase activation in starving larvae while in adults it controls lipid mobilization during flight. Larval corpora cardiaca contain about 10 times less AKH than the corpora cardiaca of adults. The corpora cardiaca of M. sexta appear to contain only one AKH.

Animals↗

A novel adipokinetic peptide in a water boatman (Heteroptera, Corixidae) and its bioanalogue in a saucer bug (Heteroptera, Naucoridae).

The corpora cardiaca (CC) of two water bug species, the water boatman Corixa punctata and the saucer bug Ilyocoris cimicoides, contain a substance that cause hyperlipemia in the migratory locust. The primary sequence of one octapeptide belonging to the adipokinetic hormone (AKH)/red pigment-concentrating hormone (RPCH) family was deduced from the multiple MS(N) electrospray mass data of CC material from each species. Whereas the saucer bug contains the known octapeptide pGlu-Val-Asn-Phe-Ser-Pro-Ser-Trp amide, code-named Anaim-AKH, the water boatman has a novel peptide identified as pGlu-Leu/Ile-Asn-Phe-Ser-Pro-Ser-Trp amide, code-named Corpu-AKH. The ambiguity about the amino acid at position 2, i.e. Leu or Ile, in Corpu-AKH was solved by isolating the peptide in a single-step by reversed-phase HPLC and establishing co-elution with the synthetic peptide containing Leu at position 2. Functionally, the peptides regulate lipid mobilization, as evidenced by an adipokinetic effect after injecting synthetic Anaim-AKH and Corpu-AKH into the respective acceptor species. Swimming activity of I. cimicoides also causes hyperlipemia.

Amino Acid Sequence↗

Impaired ABCA1-dependent lipid efflux and hypoalphalipoproteinemia in human Niemann-Pick type C disease.

The cholesterol trafficking defect in Niemann-Pick type C (NPC) disease leads to impaired regulation of cholesterol esterification, cholesterol synthesis, and low density lipoprotein receptor activity. The ATP-binding cassette transporter A1 (ABCA1), which mediates the rate-limiting step in high density lipoprotein (HDL) particle formation, is also regulated by cell cholesterol content. To determine whether the Niemann-Pick C1 protein alters the expression and activity of ABCA1, we determined the ability of apolipoprotein A-I (apoA-I) to deplete pools of cellular cholesterol and phospholipids in human fibroblasts derived from NPC1+/+, NPC1+/-, and NPC1-/- subjects. Efflux of low density lipoprotein-derived, non-lipoprotein, plasma membrane, and newly synthesized pools of cell cholesterol by apoA-I was diminished in NPC1-/- cells, as was efflux of phosphatidylcholine and sphingomyelin. NPC1+/- cells showed intermediate levels of lipid efflux compared with NPC1+/+ and NPC1-/- cells. Binding of apoA-I to cholesterol-loaded and non-cholesterol-loaded cells was highest for NPC1+/- cells, with NPC1+/+ and NPC1-/- cells showing similar levels of binding. ABCA1 mRNA and protein levels increased in response to cholesterol loading in NPC1+/+ and NPC1+/- cells but showed low levels at base line and in response to cholesterol loading in NPC1-/- cells. Consistent with impaired ABCA1-dependent lipid mobilization to apoA-I for HDL particle formation, we demonstrate for the first time decreased plasma HDL-cholesterol levels in 17 of 21 (81%) NPC1-/- subjects studied. These results indicate that the cholesterol trafficking defect in NPC disease results in reduced activity of ABCA1, which we suggest is responsible for the low HDL-cholesterol in the majority of NPC subjects and partially responsible for the overaccumulation of cellular lipids in this disorder.

ATP Binding Cassette Transporter 1↗

Lipid demixing and protein-protein interactions in the adsorption of charged proteins on mixed membranes.

The adsorption free energy of charged proteins on mixed membranes, containing varying amounts of (oppositely) charged lipids, is calculated based on a mean-field free energy expression that accounts explicitly for the ability of the lipids to demix locally, and for lateral interactions between the adsorbed proteins. Minimization of this free energy functional yields the familiar nonlinear Poisson-Boltzmann equation and the boundary condition at the membrane surface that allows for lipid charge rearrangement. These two self-consistent equations are solved simultaneously. The proteins are modeled as uniformly charged spheres and the (bare) membrane as an ideal two-dimensional binary mixture of charged and neutral lipids. Substantial variations in the lipid charge density profiles are found when highly charged proteins adsorb on weakly charged membranes; the lipids, at a certain demixing entropy penalty, adjust their concentration in the vicinity of the adsorbed protein to achieve optimal charge matching. Lateral repulsive interactions between the adsorbed proteins affect the lipid modulation profile and, at high densities, result in substantial lowering of the binding energy. Adsorption isotherms demonstrating the importance of lipid mobility and protein-protein interactions are calculated using an adsorption equation with a coverage-dependent binding constant. Typically, at bulk-surface equilibrium (i.e., when the membrane surface is "saturated" by adsorbed proteins), the membrane charges are "overcompensated" by the protein charges, because only about half of the protein charges (those on the hemispheres facing the membrane) are involved in charge neutralization. Finally, it is argued that the formation of lipid-protein domains may be enhanced by electrostatic adsorption of proteins, but its origin (e.g., elastic deformations associated with lipid demixing) is not purely electrostatic.

Adsorption↗

Unsaturated fatty acid compositional changes and desaturation during the embryonic development of the chicken (Gallus domesticus).

An investigation has been made to correlate the activities of the delta 9- and delta 6-long chain fatty acid desaturation systems with the increased levels of oleic and arachidonic acids in the liver relative to the yolk sac membrane of the chick embryo during the last week of development. The membrane exhibited high levels of both stearic and linoleic acid desaturation in the early stages of yolk lipid mobilization, the activities of both enzyme systems decreasing with the approach of hatching. Stearic acid desaturation in the liver also decreased with the approach of hatching, but linoleic acid desaturation increased. The observed levels of desaturation in the yolk sac membrane are capable of making a considerable contribution to the accumulations of mono- and polyunsaturated fatty acids in the embryonic liver, the requirement for which does not appear to be satisfied by the yolk lipids. With the approach of hatching and the functional regression of the yolk sac membrane, the role is taken over by the embryonic tissues.

Animals↗

Human adipose tissue blood flow during prolonged exercise, III. Effect of beta-adrenergic blockade, nicotinic acid and glucose infusion.

Subcutaneous adipose tissue blood flow (ATBF) was measured in six male subjects by the 133Xe-washout technique during 3-4 h of exercise at a work load corresponding to an oxygen uptake of about 1.71/min. The measurements were done during control conditions, during blockade of lipolysis by nicotinic acid, during acute i.v. beta-adrenergic blockade by propranolol, and during continuous i.v. infusion of glucose. The most pronounced lipid mobilization and utilization during work was seen in the control experiments where ATBF rose 3-fold on average from the initial rest period to the third hour of work. No increase in lipolysis and no increase in ATBF were found when lipolysis was blocked by nicotinic acid (0.3 g/h). Propranolol treatment (0.15 mg/kg) reduced lipolysis and nearly abolished the increase in ATBF during exercise. Intravenous administration of glucose (about 0.25 g/min) did not influence lipid metabolism (evaluated by the respiratory quotient) nor did it reduce the ATBF response to exercise. These results are inconsistent with the hypothesis that increase in ATBF during exercise is elicited via direct stimulation of vascular beta1-receptors, while they are not in disagreement with the hypothesis that adipose tissue vasodilation during exercise is secondary to metabolic events connected to lipolysis.

Adipose Tissue↗