Biomedical subjects

E Shrago

Publications and source records attributed to E Shrago.

At least 55 records · Page 3Linked to original sources

PubMed1985

Fatty acid activation of the reconstituted brown adipose tissue mitochondria uncoupling protein.

The effect of fatty acids, palmitoyl-CoA, and N',N-dicyclohexylcarbodiimide on the ion conductance of the reconstituted brown adipose tissue mitochondria uncoupling protein was investigated. 1, 5, and 10 microM palmitic acid induced a specific, GDP inhibited, increase in proton conductance in proteoliposomes containing the uncoupling protein but not in proteoliposomes prepared with purified protein extracts of liver mitochondria. 10 microM oleic acid, like palmitic acid, increased proton conductance in proteoliposomes prepared with the uncoupling protein. Palmitoyl-CoA and caprylic acid had no effect on increasing proton conductance. Similar to the observation in mitochondria, there was no effect of palmitic acid on Cl-conductance, but unlike mitochondria its activation by palmitoyl-CoA or inhibition by N',N-dicyclohexylcarbodiimide was lost. The results, obtained in an isolated system, provide support for the contention that long chain fatty acids act as an acute physiological activator of the uncoupling protein.

Adipose Tissue, Brown↗

PubMed1985

Dietary and nutritional aspects of fatty acid binding proteins.

Information on cytosolic fatty acid binding proteins (FABP) related to dietary and pharmacological manipulations is discussed in terms of FABP function. FABP present in liver, heart, intestinal mucosa and omental fat responds to different diets. A parallel change occurs in tissue levels of FABP and metabolism of fatty acids. It seems FABP might play a role in lipid metabolism by interacting with membrane bound enzymes. The available data also support the argument in favor of FABP involvement in intracellular transport, compartmentalization and channeling of fatty acids.

Animals↗

PubMed1985

Adenine nucleotide translocase activity and sensitivity to inhibitors in hepatomas. Comparison of the ADP/ATP carrier in mitochondria and in a purified reconstituted liposome system.

Adenine nucleotide uptake was found to be lower in mitochondria from hepatoma 7777, 7800, and 9618A than in the host livers. Moreover, in the fast-growing hepatoma 7777 the sensitivity of the adenine nucleotide translocase to inhibition by carboxyatractylate and bongkrekic acid was considerably decreased. Purification of the ADP/ATP carrier from hepatoma 7777 mitochondria and its reconstitution into an artificial liposome system reversed the abnormal kinetics in that the adenine nucleotide uptake and response to inhibitors were identical in proteoliposome preparations from host liver and tumor mitochondria. Analysis of the lipids of the hepatoma inner mitochondrial membrane indicated considerable differences from normal in the levels of phospholipids and cholesterol. Most striking was the increase in cholesterol and sphingomyelin of the hepatoma 7777 inner membrane. An artificial liposome system containing cholesterol in addition to the standard phospholipids could produce alterations in kinetics of the purified ADP/ATP carrier from heart mitochondria similar to those seen in the hepatoma 7777. In general, these results support the suggestion that alterations in the lipid environment of the inner mitochondrial membrane rather than intrinsic changes in the carrier protein itself produce the aberrant observations of adenine nucleotide translocase activity in hepatoma mitochondria.

Animals↗

PubMed1985

Regulation of aminotransferase-glutamate dehydrogenase interactions by carbamyl phosphate synthase-I, Mg2+ plus leucine versus citrate and malate.

Citrate, malate, and high levels of ATP dissociate the mitochondrial aspartate aminotransferase-glutamate dehydrogenase complex and have an inhibitory effect on the latter enzyme. These effects are opposed by Mg2+, leucine, Mg2+ plus ATP, and carbamyl phosphate synthase-I. In addition, Mg2+ directly facilitates formation of a complex between glutamate dehydrogenase and the aminotransferase and displaces the aminotransferase from the inner mitochondrial membrane which could enable it to interact with glutamate dehydrogenase in the matrix. Zn2+ also favors an aminotransferase-glutamate dehydrogenase complex. It, however, is a potent inhibitor of and has a high affinity for glutamate dehydrogenase. Leucine, however, enhances binding of Mg2+ and decreases binding of and the effect of Zn2+ on the enzyme. Thus, since both metal ions enhance enzyme-enzyme interaction and Zn2+ is a more potent inhibitor, the addition of leucine in the presence of both metal ions results in activation of glutamate dehydrogenase without disruption of the enzyme-enzyme complex. Furthermore, the combination of leucine plus Mg2+ produces slightly more activation than leucine alone. These results indicate that leucine, carbamyl phosphate synthase-I, and its substrate and cofactor, ATP and Mg2+, operate synergistically to facilitate glutamate dehydrogenase activity and interaction between this enzyme and the aminotransferase. Alternatively, Krebs cycle intermediates, such as citrate and malate, have opposing effects.

Adenosine Triphosphate↗

PubMed1985

Partial purification and functional reconstitution of GDP sensitive brown adipose tissue mitochondria uncoupling protein using octyl glucoside.

A partial purification of the uncoupling protein of brown adipose tissue mitochondria (BATM) was achieved by extraction of BATM with 40 mM octyl glucoside, followed by affinity chromatography on ATP-agarose. The isolated protein was functionally reconstituted into liposomes using octyl glucoside dialysis. Proteoliposomes containing the uncoupling protein had an increased proton or chloride conductance when subjected to a valinomycin-induced potassium diffusion potential. The increased ion conductance was consistently found to be inhibited by 200 microM GDP.

Adipose Tissue, Brown↗

PubMed1985

Carnitine palmitoyltransferase. Activation by palmitoyl-CoA and inactivation by malonyl-CoA.

Extraction of rat liver mitochondria twice with 0.5% Triton X-100 in a salt-free medium leaves less than 10% of the carnitine palmitoyltransferase membrane bound. The remaining membrane-bound enzyme is inhibited virtually completely by 10 microM malonyl-CoA. Preincubation of the extracted membranes with palmitoyl-CoA and salts (KCI) for several minutes activates the enzyme and makes it increasingly insensitive to malonyl-CoA. Addition of malonyl-CoA to the preincubation reverses this desensitization. In albumin-containing media salts also decrease the binding of palmitoyl-CoA to albumin and stimulate carnitine palmitoyltransferase by increasing substrate availability in free solution. The reverse reaction shows accelerated desensitization by palmitoylcarnitine and resensitization by malonyl-CoA.

Acyl Coenzyme A↗

PubMed1985

Extraction of tissue long-chain acyl-CoA esters and measurement by reverse-phase high-performance liquid chromatography.

Long-chain acyl-CoA esters were extracted from freeze-clamped livers of fed and fasted rats according to the method of Mancha et al. [M. Mancha, G. B. Stokes, and P. K. Stumpf (1975) Anal. Biochem. 68, 600-608] and analyzed on a radially compressed C18, 5 microns, reverse-phase column using a gradient system consisting of acetonitrile and 25 mM KH2PO4, pH 5.3, at 254 nm. Total analysis time was 25 min. Eight peaks in the extract with carbon chain lengths of 12 to 18, which subsequently disappeared on alkaline hydrolysis, were identified. The major acyl-CoA peaks in the extract in order of increasing retention times were 14:0, 16:1, 18:2, 16:0, 18:1, and 18:0. Total liver long-chain acyl-CoA esters were 108 +/- 11 and 248 +/- 19 nmol/g protein for fed and fasted rats, respectively. On fasting (48 h) the levels of 18:2, 16:0, and 18:1 increased two-to threefold and that of 18:0 sixfold. The advantages of this method are that it not only provides a more direct determination of total tissue long-chain acyl-CoA esters, in that no decomposition of the CoA ester is involved, but it also detects the constituent molecular species.

Acyl Coenzyme A↗

PubMed1985

Brown adipose tissue from fetal rhesus monkey (Macaca mulatta): morphological and biochemical aspects.

Brown adipose tissue (BAT) from fetal rhesus monkeys microscopically resembled adult rodent BAT containing multiocular fat cells with numerous mitochondria. Mitochondrial carnitine palmitoyl transferase activity was lower than that in adult rodents and adenine nucleotide translocase activity was similar to that reported for rats. Rhesus monkey BAT mitochondria (BATM) possess an uncoupling protein that is characteristic of BAT as evidenced by the binding of [3H]GDP, the inhibition by GDP of the high Cl- permeability or rapid alpha-glycerol-3-phosphate oxidation. Electrophoretic analysis of BATM showed the presence of a 32,000 mol.wt protein which was enriched by procedures established for the isolation of BATM uncoupling protein.

Adipose Tissue, Brown↗

PubMed1985

Specific interaction of fatty acyl-CoA esters with brown adipose tissue mitochondria.

The ability of low concentrations of long-chain fatty acyl coenzyme A (CoA) esters to act as inhibitors of purine nucleotide action in hamster brown adipose tissue mitochondria was observed to result from a specific interaction. Palmitoyl-CoA was found to be a competitive inhibitor of nucleotide binding with an apparent Ki of 2.46 +/- 1.09 microM for GDP and 2.98 +/- 0.538 microM for ATP and was able to counteract GDP-inhibited mitochondrial swelling. A minimum acyl-CoA carbon chain length of 12 was necessary for any significant inhibition of GDP binding or induction of swelling to be observed. The effect of palmitoyl-CoA on reversing GDP-inhibited chloride permeability of brown adipose tissue mitochondria was found to be the result of a specific interaction with the brown adipose tissue mitochondrial uncoupling protein. Mitochondria pretreated with N,N'-dicyclohexylcarbodiimide, which binds covalently to the uncoupling protein and partially inhibits brown adipose tissue mitochondrial swelling, underwent a nonspecific increase in swelling in the presence of phenylmercuric acetate. However, with palmitoyl-CoA no further increase in permeability could be mediated. In addition, under certain experimental conditions, palmitoyl-CoA was found to partially inhibit the high halide permeability of brown adipose tissue mitochondria but to a lesser extent than that observed with GDP, suggesting it may be acting as a partial agonist.

Acyl Coenzyme A↗

PubMed1985

Purification and characterization of fatty acid binding protein in mammalian lung.

A fatty acid-binding protein (FABP) has been isolated and characterized from rat lung tissue. Rat lung FABP has a slightly higher molecular weight than liver FABP, but immunologically, lung FABP is similar to that of liver FABP. Long chain acyl CoA synthetase, a key enzyme in fatty acid metabolism is stimulated by partially purified lung FABP, suggesting a physiologic role of the protein in the activation of long chain fatty acids in pulmonary tissue.

Animals↗

PubMed1985

Diet and exercise training effects on resting metabolic rate.

The decline in resting metabolic rate (RMR) during periods of caloric restriction is a well documented phenomenon. The purpose of this study was to determine if either 30 min of daily self-selected aerobic activity (Group D) or prescribed exercise training performed every other day (Group P) for 12 weeks can prevent the decrease in RMR during caloric restriction for weight loss. Seventy-eight adult subjects (38 M, 40 F) whose weights were 15-35 percent above the upper limit for age, sex and frame were randomly assigned to three treatment groups. All three groups followed the same 1200, 1500 or 1800 kcal/d (5023, 6279, 7534 kJ) diet exchange plan. Group C (Control) followed no exercise program, while D and P exercised as described above. Maximal oxygen consumption (VO2 max, m10(2)/kg X min-1) predicted from the Bruce Test increased by 0.08 +/- 7, 9 +/- 12 and 12 +/- 9 percent in groups C, D and P, respectively. The increases for D and P were significantly higher than for C (P less than 0.01). RMR (m10(2)/kg X min-1) increased by 2 +/- 9, 4 +/- 7 and 10 +/- 9 percent in groups C, D and P respectively. The percent change for group P was significantly greater than that for groups C and D (P less than 0.05). There was a significant relationship across all subjects between the % delta in VO2 max and % delta in RMR (r = 0.307, P less than 0.01). However, the association between these two variables was stronger for females than for males.(ABSTRACT TRUNCATED AT 250 WORDS)

Adult↗

PubMed1984

Decreases in serum high-density-lipoprotein cholesterol and total cholesterol resulting from naturally produced and recombinant DNA-derived leukocyte interferons.

A three-phase study was conducted to examine the effect of leukocyte interferon administration on serum high-density-lipoprotein (HDL) cholesterol and total cholesterol levels. In the initial phase, human leukocyte interferon decreased HDL cholesterol (P less than 0.05) and total cholesterol (P less than 0.05) levels in patients with breast carcinoma. Decreases began with initiation of the interferon administration, were sustained throughout the period of treatment, and increased toward pretreatment values with discontinuation of interferon. In the second phase of the study, in neither of two comparison groups of women receiving cytotoxic chemotherapy, excluding interferon, did any similar decrease in HDL cholesterol occur. In a third comparison group of women being treated for metastatic breast carcinoma, a predicted and significant (P less than 0.01) drop in HDL cholesterol level without a concomitant lowering of total cholesterol level occurred immediately following the initiation of androgen therapy. To confirm that the observed cholesterol level decreases were associated with interferon rather than a contaminant thereof, analyses were also carried out on samples from a study utilizing interferon rather than a contaminant thereof, analyses were also carried out on samples from a study utilizing interferon produced by recombinant DNA techniques and purified to homogeneity. A similar decrease in HDL cholesterol (P less than 0.05) and total cholesterol (P less than 0.01) was observed. A definite relationship therefore appears to exist between the administration of human leukocyte interferon and decreased plasma levels of HDL cholesterol and total cholesterol.

Aged↗

PubMed1983

An adenine nucleotide-phosphoenolpyruvate counter-transport system in C3 and C4 plant chloroplasts.

A rapid counter-exchange between ATP and phosphoenolpyruvate (PEP) has been demonstrated in pea and maize mesophyll chloroplasts. Chloroplasts preloaded with either [14C] ATP or [14C] PEP readily exchange the radioactive compound with the externally added anions, ATP or PEP, whereas, cold external Pi counter-transports only with internal [14C] PEP. Flooding the system with cold Pi, however, will significantly reduce the counter-transport of external cold PEP with internal [14C] ATP. This ATP-PEP exchange is also markedly decreased by lowering the incubation temperature. The results indicate that the ATP-PEP counter-exchange could represent a key transport system in plant chloroplasts and may be particularly important in the photosynthesis of C4 plants. Furthermore, they provide information required to elucidate the mechanism of the ATP-PEP counter-transport system.

Adenosine Triphosphate↗

PubMed1983

Effect of thyroidectomy on the kinetics of ADP-ATP translocation in liver mitochondria.

The role of adenine nucleotide translocase (AdNT) in the reduced oxidative metabolism of hypothyroidism has been examined. Both AdNT and respiratory activities in liver mitochondria of thyroidectomized rats were 30% below normal. Mitochondrial AdNT activities were determined by the back-exchange method of Pfaff and Klingenberg (Eur. J. Biochem. 6, 66, 1968). The Km and Vmax of the enzyme were temperature dependent. At physiological temperature, the Km and Vmax of the normal rat AdNT were 10 microM (for external ADP) and 4.73% s-1 (percentage efflux of the labeled adenine nucleotides), respectively. AdNT in hypothyroid rat liver mitochondria exhibited a 25-35% lower Vmax and 75% higher Km when assayed over the temperature range 0 to 37 degrees C. Dixonplot studies indicated that the AdNT in hypothyroidism was two- to threefold more sensitive to atractylate and palmitoyl-CoA inhibitions. In contrast the ADP-ATP translocase in hypothyroidism was more resistant than the control carrier to bongkrekate inhibition. The decrease in the transport of ADP, which is consistent with the decreased oxidative activity associated with hypothyroidism, apparently occurs secondary to changes in the lipid matrix of the inner mitochondrial membrane (F. L. Hoch (1977) Arch. Biochem. Biophys. 178, 535.).

Adenosine Diphosphate↗

PubMed1983

Modification of liver mitochondrial lipids and of adenine nucleotide translocase and oxidative phosphorylation by cold adaptation.

The decrease in respiration rate following thyroidectomy is preceded by changes in the lipid composition of the mitochondrial membrane (Hoch, F.L., Subramanian, C., Dhopeshwarkar, G.A. and Mead, J.F. (1981) Lipids 16, 328-334) and in concert, changes in the kinetic parameters of the adenine nucleotide translocase (Mak, I.T., Shrago, E. and Elson, C.E. (1981) Fed. Proc. 40, 398). To demonstrate that physiological adaptation also involves this sequence of events, rats were housed at 8 degrees C for 3-4 weeks. Cold adaptation resulted in a modest (5%) increase in the unsaturation index for the mitochondrial fatty acids comprised of a significant increase in arachidonic acid and a reciprocal decrease in linoleic acid. Phospholipid analysis indicated that cold adaptation increased the mitochondrial phosphatidylethanolamine and reciprocally decreased the phosphatidylcholine content. Concomitantly, cold adaptation resulted in 25-30% increases in rat liver mitochondrial respiratory activities without changing the respiratory control or ADP/O ratios. The kinetic parameters of the adenine nucleotide translocase were determined by the back-exchange method (Pfaff, E. and Klingenberg, M. (1968) Eur. J. Biochem. 6, 66-79). At 0-4 and 10 degrees C, the Vmax and Km of the cold-adapted rat liver adenine nucleotide translocase were not distinguishable from the control values. The Ki values determined by Dixon plot studies for atractylate and palmitoyl-CoA were also comparable between the two groups. However, at 25 and 37 degrees C, cold-adapted rat liver adenine nucleotide translocase exhibited a 20% increase in Vmax and a 20% decrease in Km for external ADP. The results suggest that one adaption to a cold environment involves hormone-mediated changes in the lipid composition in the mitochondrial membranes which in turn modulate the adenine nucleotide translocase and subsequent respiratory activities.

Acclimatization↗

PubMed1983

The influence of linoleic acid intake on the kinetics of adenine nucleotide translocase.

Dietary enrichment of mitochondrial polyenoic fatty acid content was associated with increased respiratory activity (Abuirmeileh and Elson). The influence of the membrane lipid composition on the adenine nucleotide translocase (AdNT) was studied in rats that were fed diets formulated with beef tallow (BT) to provide low or safflower oil (SO) to provide high contents of linoleic acid. The phosphatidylcholine/phosphatidylethanolamine ratio was 40% higher in the SO mitochondria due primarily to an increase in phosphatidylcholine. SO mitochondria exhibited 25% higher state 3 respiration, 50% higher state 4 respiration and 13% higher net ADP-dependent respiration than did the BT mitochondria. The relative RCR and ADP/O values of the SO mitochondria were slightly but significantly (p less than 0.01) lower. The kinetics of the AdNT were determined by the back exchange method (Pfaff and Klingenberg). At all assay temperatures, SO mitochondria exhibited a higher Km for ADP. However, addition of 5 mM carnitine to the assay mixture increased the affinity of the SO-AdNT giving Km values similar to that of the BT-AdNT. Washing the mitochondria with fat-free BSA had a similar, but lesser, effect. At 25 C and 37 C, the Vmax of the SO-AdNT were increased by 11-15% (p less than 0.05) which was independent of either BSA-wash or the presence of carnitine. According to Dixon plot studies, the SO-AdNT had a comparable Ki for atractylate but a slightly lower Ki for palmitoyl-CoA inhibition. The accumulation of acyl-CoA esters in the SO mitochondria was not ruled out. The overall results suggest that changes in mitochondrial membrane lipids accommodated an increased Vmax of the SO-AdNT, which in turn accounted for that part of the increased state 3 respiration dependent on ADP translocation.

Animals↗

PubMed1983

Total cholesterol and HDL-cholesterol changes during acute, moderate-intensity exercise in men and women.

Chronic endurance exercise training has been associated with decreased levels of total cholesterol and increased HDL-cholesterol. To our knowledge rapid changes in cholesterol and HDL-cholesterol during acute exercise have not been described under controlled conditions. We studied 28 subjects (14 males and 14 females) during bicycle exercise for 40 min at a work intensity of 55% of their maximal oxygen consumption. Total and HDL-cholesterol levels were measured (and LDL-cholesterol calculated) at rest, 10, 20, 30, and 40 min of exercise, and 15 min postexercise. There was a significant (p less than 0.001) increase in HDL-cholesterol levels at 10 min of exercise (58.8 +/- 13.9 mg/dl, mean +/- SD) above rest (53.1 +/- 13.4 mg/dl) for all subjects. This increase persisted (p less than 0.001) at all time points throughout the exercise session, but declined by 15 min postexercise. There was a small, insignificant decline in LDL-cholesterol. It is concluded that apparent favorable changes in lipoprotein patterns occur acutely, and are sustained during short-term, moderate intensity exercise. Analyses of these changes appears necessary if the biochemical mechanisms which underlie these metabolic alterations are to be elucidated.

Adolescent↗

PubMed1983

Effects of acute moderate-intensity exercise on carnitine metabolism in men and women.

The purpose of this investigation was to describe the dynamics of carnitine metabolism during an acute episode of exercise. Twenty-eight subjects (14 male; 14 female) exercised for 40 min on a bicycle ergometer at 55% of their maximal aerobic capacities. Blood samples were obtained at rest, 10, 20, 30, and 40 min of exercise, and 15-min postexercise. Muscle biopsies of the vastus lateralis were performed before and after exercise. Results demonstrated that the percent of acylated plasma carnitine increased significantly (P less than 0.05) across all subjects from 17.3% at rest to 22.3% by 40 min of exercise and continued to increase to 22.8% 15-min postexercise. Total muscle carnitine levels fell significantly (P less than 0.001) across all subjects from 4.21 (1.27) (means +/- SD) mumol/g wet weight at rest to 3.29 (1.27) mumol/g wet weight after exercise. Well-trained males and females had almost identical levels of muscle carnitine [4.35(1.86) and 4.34 (0.64) mumol/g wet weight, respectively]. These levels were somewhat higher but not significantly higher than their moderately trained counterparts [3.86(1.34) and 4.28(1.18) males and females, respectively]. Carnitine palmitoyl transferase (E.C. 2.3.1.21) activity also declined significantly (P less than 0.05) across all subjects after exercise. This study is the first to demonstrate a potential loss of acylated carnitine forms from muscle to plasma during acute exercise, possibly reflecting an increase in carnitine turnover. Alterations in carnitine status may represent another metabolic adaptation to chronic exercise training.

Adult↗