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G Dallner

Publications and source records attributed to G Dallner.

At least 37 records · Page 2Linked to original sources
PubMed1998

Analysis of ubiquinone and tocopherol levels in normal and hyperlipidemic human plasma.

The type and amount of lipophilic antioxidants in plasma of hyperlipidemic patients are of great importance since they play a central role in preventing deleterious oxidation of blood lipids and proteins. Isolation and quantitation of lipophilic antioxidants from hyperlipidemic plasma samples meet great obstacles because of increased levels of various intermediary lipid products. This study was designed to develop a rapid and efficient extraction and separation procedure for simultaneous analysis of ubiquinone-9 and -10 as well as alpha-, delta-, and gamma-tocopherol isomers. The levels of ubiquinone-10, alpha- and gamma-tocopherol were analyzed in human plasma samples using high-performance liquid chromatography. Lipid extraction was performed by petroleum ether/methanol/water. After phase separation, ubiquinone was reduced to ubiquinol by sodium borohydride and the lipids were separated on a C18 column. A binary gradient with solvents containing lithium perchlorate was used, and an electrochemical detector was employed for quantitation. This procedure was also efficient for the analysis of antioxidant lipids in samples containing a large number of accumulated and interfering lipid intermediates. Thus, the procedure described here is useful for efficient and rapid quantitation of ubiquinones and tocopherols in human plasma samples, especially those originating from hyperlipidemic patients.

Chromatography, High Pressure Liquid↗
PubMed1998

Gemfibrozil-induced decrease in serum ubiquinone and alpha- and gamma-tocopherol levels in men with combined hyperlipidaemia.

BACKGROUND: Low blood levels of antioxidants are associated with an increased risk of developing coronary artery disease. Lipophilic antioxidants are transported in lipoproteins, and hypolipidaemic therapy may therefore alter their blood concentrations. METHODS: The present randomized, placebo-controlled cross-over study of 21 men with combined hyperlipidaemia examines whether 10-12 weeks of gemfibrozil treatment affects the serum concentrations of the antioxidants ubiquinone-10 or alpha- or gamma-tocopherol. RESULTS: Gemfibrozil treatment lowered plasma triglycerides and both total and very low-density lipoprotein (VLDL)-cholesterol (P < 0.001 for all by ANOVA), whereas high-density lipoprotein (HDL)-cholesterol increased (P < 0.001). The median serum levels of ubiquinone-10 decreased from 1.30 mumol L-1 (interquartile range 0.87-1.71 mumol L-1) with placebo to 0.76 mumol L-1 (0.66-0.95) with gemfibrozil treatment (P < 0.001). Corresponding levels for alpha- and gamma-tocopherol were: 68.5 mumol L-1 (51.1-84.7) vs. 40.8 mumol L-1 (30.3-55.0) and 8.6 mumol L-1 (5.2-16.7) vs. 4.3 mumol L-1 (3.5-7.0) respectively (P < 0.001 for both). The decrease in serum antioxidants was also evident when standardized for total cholesterol (P < 0.05) or LDL-cholesterol (P < 0.001). Normolipaemic control subjects had significantly lower antioxidant levels than placebo-treated patients: ubiquinone 0.63 mumol L-1 (0.41-1.05), alpha-tocopherol 34.3 mumol L-1 (27.3-45.6) and gamma-tocopherol 3.2 mumol L-1 (2.5-4.2) (P < 0.001 for all). The association of antioxidants with lipoprotein lipids was further established by positive correlations between the levels of antioxidants and those of total cholesterol (r = 0.64, P < 0.001) or total triglycerides (r = 0.71, P < 0.001). CONCLUSION: Gemfibrozil treatment of men with combined hyperlipidaemia reduces serum antioxidant levels to the levels seen in healthy normolipidaemic men. The mechanisms and the relevance of this finding remain unclear and need to be addressed in further studies.

Adult↗
PubMed1997

Chloroplastic prenylated proteins.

By in vivo [3H]mevalonate labelling of spinach combined with biochemical analysis, evidence is provided for the existence of protein prenylation in chloroplasts. Approximately 20 prenylated polypeptides were resolved by SDS-PAGE followed by autoradiography. Thermolysin treatment of intact chloroplasts revealed that about 40% of the prenylated polypeptides were associated with the cytoplasmic surface of the outer envelope membrane. The remaining portion was present in thylakoids and/or the inner envelope membrane. The majority of the prenylated polypeptides were associated with larger membrane protein complexes. A farnesyl protein transferase activity was found to be associated with the thylakoid membrane.

Alkyl and Aryl Transferases↗
PubMed1997

Peroxisomal impairment in Niemann-Pick type C disease.

Niemann-Pick type C disease (NPC) belongs to the group of lysosomal storage diseases characterized by an accumulation of cholesterol and sphingomyelin. Using a mutant mouse strain, enzymatic markers for lysosomes, mitochondria, microsomes, and peroxisomes were investigated in the liver and brain. Aside from lysosomal changes, we found a sizable decrease of peroxisomal beta-oxidation of fatty acids and catalase activity in the brain and liver. Isolated peroxisomes displayed a significant decrease of these enzyme activities. Furthermore, the only phospholipid change in brain was a decreased content of the plasmalogen form of phosphatidylethanolamine, and the dimethylacetal pattern was also modified. The electron microscopical appearance of peroxisomes did not display any large changes. The defect of peroxisomal enzymes was already present 18 days before the onset of the disease. In contrast, the lysosomal marker enzyme increased in activity only 6 days after appearance of the symptoms. The events of the studied process have previously been considered to be elicited by a lysosomal deficiency, but this study demonstrates disturbances similar to those in a number of peroxisomal diseases. It appears that the peroxisomal impairment is an early event in the process and could be a factor in the development of Niemann-Pick type C disease.

Acid Phosphatase↗
PubMed1997

Protein prenylation in spinach--tissue specificity and greening-induced changes.

Etiolated spinach seedlings, as well as petioles and blades of leaves of green seedlings, were labeled with [3H]mevalonate to study protein prenylation in several plant developmental stages. The polypeptide prenylation pattern of the leaf petiole and the leaf blade differed considerably, although some prenylated proteins were present in both tissues. During greening several prenylated polypeptides in the 30- to 46-kDa molecular mass region and two at 15 kDa became more abundant, while others in the 21.5- to 30-kDa region and one at 62 kDa showed a relative decrease. However, the relative amount of several of the prenylated polypeptides did not appear to be altered during the greening process. In etiolated seedlings, more thioether-linked farnesol than geranylgeraniol was found, whereas in seedlings grown under normal light conditions the converse situation prevailed.

Chlorophyll↗
PubMed1997

Lipid peroxidation and changes in the ubiquinone content and the respiratory chain enzymes of submitochondrial particles.

The relationship between, lipid peroxidation induced by ascorbate and adenosine ADP/Fe3+, and its effect on the respiratory chain activities of beef heart submitochondrial particles has been investigated. Lipid peroxidation, measured as thiobarbituric acid reactive substance formation, resulted in an inhibition of the NADH and succinate oxidase activities. Examination of several partial reactions of the respiratory chain revealed inactivation primarily of those involving endogenous ubiquinone, i.e., NADH- and succinate-ubiquinone1 and cytochrome c reductases. Ubiquinol-cytochrome c reductase, measured with reduced ubiquinone2 as electron donor, was unaffected. The amount of NADH- or succinate-reducible cytochrome b in the presence of cyanide was strongly decreased, but could be recovered by the addition of antimycin. There occurred a substantial decrease of the ubiquinone content in the course of lipid peroxidation, with a linear relationship between this decrease and the NADH and succinate oxidase activities. The results are consistent with the conclusion that the ubiquinone pool undergoes an oxidative modification during lipid peroxidation, to a form that can no longer function as a component of the respiratory chain. Lipid peroxidation also led to a partial inhibition of the succinate dehydrogenase and cytochrome c oxidase activities and a minor decrease of the cytochrome c and cytochrome a contents. Reduction of endogenous ubiquinone prevented lipid peroxidation as well as the concomitant modification of ubiquinone and inactivation of the respiratory chain. These observations suggest that the destruction of ubiquinone through lipid peroxidation is the primary cause of inactivation of the respiratory chain, and emphasize the antioxidant role of ubiquinol in preventing these effects. The possible implications of these findings for regulation of the cellular turnover of ubiquinone by the prevailing oxidative stress are discussed.

Adenosine Diphosphate↗
PubMed1996

Oxidative modification of nicotinamide nucleotide transhydrogenase in submitochondrial particles: effect of endogenous ubiquinol.

The present paper describes the sensitivity of the mitochondrial nicotinamide nucleotide transhydrogenase (EC 1.6.1.1) to oxidative modification, and the effects of endogenous ubiquinol on this modification. A comparison is made between the effects of treatment with ADP-Fe3+ and ascorbate and with peroxynitrite, using kinetic, electrophoretic, and immunological analyses, together with lipid peroxidation measurements. The transhydrogenase was inactivated by both types of oxidative modification, but apparently through different mechanisms. Ubiquinol protected the enzyme against inactivation only when the modification was caused by ADP-Fe3+ and ascorbate treatment. Kinetic measurements revealed a threefold increase of the Km value of the enzyme for NADPH after exposure to ADP-Fe3+ and ascorbate, and a twofold increase of the Km values for both NADH and NADPH after exposure to peroxynitrite. NAD(H) exerted a protection against trans-hydrogenase inactivation when added to the preincubation in the case of peroxynitrite, but neither NAD(H) or NADP(H) protected in the case of ADP-Fe3+ and ascorbate. Using immunoblotting it was shown that the enzyme became both aggregated and fragmented, although to different extents, depending on the oxidative system used. Again, ubiquinol prevented these effects only in the case of ADP-Fe3+ and ascorbate treatment. Furthermore, there occurred a striking decrease in the 66-kDa trypsin fragment after exposure of the enzyme to ADP-Fe3+ and ascorbate, and of the 48-kDa trypsin fragment after exposure to peroxynitrite. It is concluded that the mitochondrial nicotinamide nucleotide transhydrogenase is sensitive to oxidative stress and that the mechanism underlying this can vary according to the challenge to which the enzyme is exposed. Endogenous ubiquinol may play a role in protecting the enzyme against agents perturbing the lipid phase of the membrane.

Adenosine Diphosphate↗
PubMed1996

Increases in tissue levels of ubiquinone in association with peroxisome proliferation.

Rats were treated with various peroxisome proliferators and concomitant changes in ubiquinone levels were monitored. In addition to clofibrate and di(2-ethylhexyl)phthalate, acetylsalicylic acid, 2-ethylhexanoic acid, thyroxine and dehydroepiandrosterone were used as proliferators. Administration of these compounds increased the contents of ubiquinone in liver and, to some extent, in kidney and muscle. No change in corresponding valued for heart or brain were observed. The treatments did not influence cholesterol levels, but increased the amounts of dolichol in the liver to various extents. Treatment of rats with the catalase inhibitor aminotriazole increased the ubiquinone levels in kidney, heart and muscle but not in liver. Comparison of peroxisomal fatty acid beta-oxidation with ubiquinone amounts in liver homogenates after treatment with a number of peroxisome proliferators demonstrated a direct correlation between these two parameters. Subcellular fractionation of liver after peroxisome proliferation revealed that the ubiquinone level was increased in mitochondria and lysosomes which are the main compartments for this lipid, but an increase was also observed in both peroxisomes and microsomes. The increase in hepatic ubiquinone after treatment with various types of proliferators was related to the decrease in blood cholesterol level. These results show that the volume of the peroxisomal compartment and the ubiquinone content in animal tissues are interrelated.

Amitrole↗
PubMed1996

Lipid composition in scrapie-infected mouse brain: prion infection increases the levels of dolichyl phosphate and ubiquinone.

The neutral and phospholipid composition of mouse brain infected with scrapie prions was investigated. During the later stages of this disease, the level of dolichol decreased by 30% whereas the level of dolichyl phosphate increased by 30%. In terminally ill mice, there was also a 2.5-fold increase in both total ubiquinone and its reduced form. Furthermore, alpha-tocopherol was elevated at this stage by 50%. In contrast, no changes were observed in phospholipid amount, in phospholipid composition, and in phosphatidylethanolamine plasmalogen content during the entire disease process. The fatty acid and aldehyde composition of individual phospholipids remained unaltered as well. No modifications could be detected in cholesterol content. Thus, the majority of membrane lipids in scrapie-infected mouse brain are modified in neither quantity nor structure, but specific changes occur to a few polyisoprenoid lipids. This specificity indicates that, although prions accumulate in lysosomes, the infection process is not associated with a general membrane destruction caused by lysosomal enzyme leakage.

Animals↗
PubMed1996

Influence of dolichol on microsomal membrane functions.

Microsomal membranes from rat liver were extracted with n-pentane in order to remove the lipid products of the mevalonate pathway, dolichol, ubiquinone and cholesterol. Dolichol and cholesterol were subsequently reincorporated into these extracted membranes. Electron microscopic examination demonstrated that extraction did not alter the vesicular membrane structure of the microsomes. The extracted vesicles were permeable to uncharged molecules in the same manner as control microsomes but had an increased permeability for charged molecules. Enzyme denaturation was not observed. The contraction of extracted vesicles was greatly increased when the incubation medium was supplemented with non-penetrating compounds such as polyethylene glycol and was much greater than that of control microsomes. When extracted membranes were reconstituted with dolichol or cholesterol, the original lower degree of contraction was reestablished. The effects of dolichol reincorporation on a number of microsomal enzyme activities were investigated and some limited changes were observed. These results demonstrate that extraction of microsomes with n-pentane and subsequent reincorporation of dolichol is an effective approach for investigating the functions of this lipid. Dolichol and cholesterol both affect microsomal membrane fluidity, but only cholesterol modifies the activities of certain integral microsomal membrane enzymes to a larger extent.

Animals↗
PubMed1995

Identification of spinach farnesyl protein transferase. Dithiothreitol as an acceptor in vitro.

Spinach seedlings were found to contain farnesyl protein transferase. The enzyme is activated by Zn2+, but not by Mg2+. The pH optimum is approximately 7.0 and maximal activity is obtained at 40-45 degrees C. The apparent Km for the farnesyl diphosphate substrate is 7 microM. Western blotting of soluble proteins with an antiserum raised against mammalian farnesyl protein transferase demonstrated a specific cross-reactivity with the spinach enzyme. The antiserum preferentially recognises the beta-subunit of the heterodimeric farnesyl protein transferase, and the corresponding spinach polypeptide has a molecular mass of 42 kDa on SDS/PAGE. The enzyme can employ dithiothreitol as an acceptor for the farnesyl moiety and catalyses the formation of a thioether linkage between these substrates. On the basis of this discovery, a new method was developed utilising the hydrophobicity of the reaction product, and its interaction with poly(propylene). During in vivo labelling, the plants took up dithiothreitol, which inhibited the incorporation of [3H]mevalonate metabolites into proteins, indicating that dithiothreitol might be isoprenylated in vivo as well as in vitro. However, isoprenylation of some proteins remains unaffected by dithiothreitol suggesting the existence of different isoprenylation mechanisms. Thus, it is demonstrated that plants possess farnesyl protein transferase, which resembles its mammalian and yeast homologues.

Alkyl and Aryl Transferases↗
PubMed1995

Biochemical, physiological and medical aspects of ubiquinone function.

This presentation is a brief review of current knowledge concerning some biochemical, physiological and medical aspects of the function of ubiquinone (coenzyme Q) in mammalian organisms. In addition to its well-established function as a component of the mitochondrial respiratory chain, ubiquinone has in recent years acquired increasing attention with regard to its function in the reduced form (ubiquinol) as an antioxidant. Ubiquinone, partly in the reduced form, occurs in all cellular membranes as well as in blood serum and in serum lipoproteins. Ubiquinol efficiently protects membrane phospholipids and serum low-density lipoprotein from lipid peroxidation, and, as recent data indicate, also mitochondrial membrane proteins and DNA from free-radical induced oxidative damage. These effects of ubiquinol are independent of those of exogenous antioxidants, such as vitamin E, although ubiquinol can also potentiate the effect of vitamin E by regenerating it from its oxidized form. Tissue ubiquinone levels are regulated through the mevalonate pathway, increasing upon various forms of oxidative stress, and decreasing during aging. Drugs inhibiting cholesterol biosynthesis via the mevalonate pathway may inhibit or stimulate ubiquinone biosynthesis, depending on their site of action. Administration of ubiquinone as a dietary supplement seems to lead primarily to increased serum levels, which may account for most of the reported beneficial effects of ubiquinone intake in various instances of experimental and clinical medicine.

Animals↗
PubMed1995

Relationship between the endoplasmic reticulum-Golgi membrane system and ubiquinone biosynthesis.

The involvement of the various segments of the endoplasmic reticulum (ER)-Golgi system in ubiquinone biosynthesis in rat liver was investigated using subcellular fractionation. In addition to preparing rough (R) and smooth microsomes and three different Golgi fractions, a procedure was developed to isolate a smooth vesicle fraction, designated as smooth II (SII) microsomes. The electron micrographs, chemical composition, distribution of marker enzymes, pattern of glycosidases and glycosyltransferases and participation in cholesterol transport suggest that the vesicle components of this latter fraction are intermediary between the endoplasmic reticulum and Golgi system. Both R and smooth I (SI), but not SII microsomes nor Golgi vesicles demonstrate trans-prenyltransferase activity, which synthesizes the side-chain of ubiquinone from geranyl pyrophosphate (GPP). The subsequent enzyme, which transfers solanesyl pyrophosphate (sol-PP) to 4-hydroxybenzoate, is absent from R and SI microsomes, but present in SII microsomes and exhibits high levels of activity in all of the Golgi fractions. Thus, ubiquinone is synthesized sequentially in the ER-Golgi system and thereafter translocated from this compartment to other cellular membranes.

Animals↗
PubMed1995

Estimation of dolichol and cholesterol synthesis in microsomes and peroxisomes isolated from rat liver.

The participation of peroxisomal and microsomal fractions from rat liver in dolichol and cholesterol synthesis was investigated using marker enzymes. Recovery was 8% for peroxisomes and 33% for microsomes, with virtually no cross-contamination between these fractions. Using these data, it was calculated that the peroxisomal branch-point enzyme activities for dolichol and cholesterol biosynthesis, i.e. cis-prenyltransferase and squalene synthase, were 25% and 12%, respectively, of the total homogenate activity. Treatment with mevinolin increased the peroxisomal contribution in the case of both enzymes, to levels almost equal to that of their microsomal counterparts. These results indicate that peroxisomes play a role in the biosynthesis of isoprenoid lipids and that the extent of this participation is increased extensively when peroxisomes are induced by various treatments.

Animals↗
PubMed1995

Isoprenylation of plant proteins in vivo. Isoprenylated proteins are abundant in the mitochondria and nuclei of spinach.

Protein isoprenylation in vivo is demonstrated using spinach seedlings labeled with [3H]mevalonate. This report provides evidence for the occurrence of a large number of isoprenylated proteins in plants. Seedlings, without roots, were labeled quantitatively through the cut stem. Mevinolin treatment of the seedlings resulted in increased incorporation of radiolabel into proteins. Approximately 30 labeled bands could be detected after autoradiography of SDS-polyacrylamide gel electrophoresis-separated polypeptides, ranging in molecular mass from 6 to 200 kDa. Methyl iodide hydrolysis resulted in the release of covalently bound farnesol, geranylgeraniol, phytol, and some unidentified isoprenoid compounds from mevalonate-labeled proteins. It was found that all cellular fractions contained some isoprenylated proteins, although most were located in the mitochondria and nuclei. Subfractionation of the nucleus revealed that the majority of isoprenylated proteins in this compartment were components of the nuclear matrix. The results demonstrate that in vivo labeling of a complex organism can be performed using a plant system in order to study protein isoprenylation and distribution of modified proteins in different cellular compartments.

Cell Nucleus↗
PubMed1995

Endogenous ubiquinol prevents protein modification accompanying lipid peroxidation in beef heart submitochondrial particles.

This article is a study of the relationship between lipid peroxidation and protein modification in beef heart submitochondrial particles, and the protective effect of endogenous ubiquinol (reduced coenzyme Q) against these effects. ADP-Fe3+ and ascorbate were used to initiate lipid peroxidation and protein modification, which were monitored by measuring TBARS and protein carbonylation, respectively. Endogenous ubiquinone was reduced by the addition of succinate and antimycin. The parameters investigated included extraction and reincorporation of ubiquinone, and comparison of the effect of ubiquinol with those of various antioxidant compounds and enzymes, as well as the iron chelator EDTA. Under all conditions employed there was a close correlation between lipid peroxidation and protein carbonylation, and the inhibition of these effects by endogenous ubiquinol. SDS-PAGE analysis revealed a differential effect on individual protein components and its prevention by ubiquinol. Conceivable mechanisms behind the observed oxidative modifications of membrane phospholipids and proteins and of the role of ubiquinol in preventing these effects are considered.

Animals↗
PubMed1995

Coenzymes Q9 and Q10 in skeletal and cardiac muscle in tumour-bearing exercising rats.

Physical exercise increases metabolic rate, and induces both adaptational biogenesis of mitochondria in skeletal muscle and an increase in antioxidant capacity. The onset of experimental anorexia and cachexia can be delayed by voluntary exercise. As skeletal muscle is the main target for cancer cachexia, we determined the levels of coenzymes Q9 and Q10 in skeletal muscle from tumour-bearing exercising rats, and compared them to those of sedentary tumour-bearers and controls. Both tumour-bearing groups had increased levels of coenzymes Q9 and Q10 in the anterior tibial muscle (P < 0.05 for exercised animals). In the soleus muscle, only the tumour-bearing exercising animals demonstrated an increase in the levels of both coenzymes (P < 0.05). In cardiac muscle, the presence of tumour and exercise reduced the levels of coenzymes below that of sedentary controls. Exercise counteracted the anaemia in the tumour-bearing host (P < 0.05). In conclusion, the increase in antioxidant capacity in skeletal muscle indicates a defence mechanism in the tumour-bearing hosts which is augmented by physical exercise.

Animals↗