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[The effect of carbocromen on the respiratory function of rat heart mitochondria].

Rat heart mitochondria were isolated by a slightly modified method of Sordahl. The influence of carbocromen (Intensaín) on the respiratory function of the tightly coupled or uncoupled mitochondria was tested in the presence of the respiratory substrates pyruvate, alpha-ketoglutarate, D,L-palmitoylcarnitine, L-palmitoylcarnitine, and D,L-acetylcarnitine. Carbocromen (0.089--0.89 mmol/l) did not change the P/O ratio, but reduced the rate of oxygen consumption (QO2) of the coupled mitochondria by about 20% when palmitoylcarnitine was used as respiratory substrate. With ADP in excess (5 mmol/l) carbocromen decreased QO2 in state ST3 by about 35%. This drug induced reduction of QO2 is still much more evident in uncoupled mitochondria (18% with 0.045 mmol/l carbocromen, 70% with 0.89 mmol/l carbocromen). In the presence of pyruvate as substrate the respiratory function of the coupled and uncoupled mitochondria remained nearly unchanged after adding the drug. It is discussed that carbocromen could interact with the complex enzyme dependent transport of long-chain fatty acids through the mitochondrial membrane.

Adenosine Diphosphate

Studies on the ATPase complex from beef-heart mitochondria. I. Isolation and characterization of an oligomycin-sensitive and an olgiomycin-insensitive ATPase complex from beef-heart mitochondria.

1. A new method for the isolation of the oliogomycin-sensitive ATPase from beef-heart mitochondria is described. 2. A Triton-soluble ATPase complex was isolated as a by-product of the standard procedure, or as the main product when the submitochondrial particles were pretreated with 1% Triton. The ATPase activity of this complex is sensitive neither to oligomycin nor to dicyclohexylcarbodiimide. 3. The ATPase activity of the oligomycin-sensitive ATPase complex is nearly completely dependent on added phospholipids. The highest activation was found with asolectin. 4. The oligomycin-sensitive complex can be integrated into phospholipid vesicles resulting in an ATP- and Mg2+-dependent energization of the vesicles as monitored with the fluorescent dye 9-amino-6-chloro-2-methoxyacridine. 5. Aurovertin-binding studies based on fluorescence measurement reveal the presence of 1.5 mumol aurovertin-binding sites per g protein for the oligomycin-sensitive complex and about 2.2 mumol for the oligomycin-insensitive complex. 6. The preparation of the oligomycin-sensitive complex contains at least 6--7 polypeptides in addition to those derived from F1. One of these polypeptides, with an apparent molecular weight of 31 000, is virtually absent from the oligomycin-insensitive complex. 7. Some of these polypeptides have been identified and isolated.

Adenosine Triphosphatases

Shark heart mitochondria: effects of external osmolality on respiration.

Shark mitochondrial respiration was studied in media with osmolalities between 160 and 1500 milliosmoles. The respiratory control ratio, a marker for functional integrity of the isolated mitochondria, was maximal at 1000 millismoles and decreased during hypotonic or hypertonic exposure. Shark mitochondria function best at their native tonicity, a value that produces abnormal function in mammalian mitochondria.

Adenosine Diphosphate

Swelling and contraction of heart mitochondria suspended in ammonium phosphate.

Bovine heart mitochondria which have been allowed to swell in isotonic NH4+ phosphate contract in response to initiation of oxidative phosphorylation. The contraction occurs optimally at pH 6.0 and appears from inhibition studies to result from Pi uptake being slower than removal of internal Pi via phosphorylation of external ADP. Similar results are obtained when K+ + nigericin is substituted for NH4+. Mersalyl inhibition of Pi transport in respiring, nonphosphorylating mitochondria which have been allowed to swell in NH4+ phosphate reveals a contractile process having an alkaline pH optimum. This contraction resembles closely the contraction observed in salts of strong acids and presumably occurs by electrophoretic ejection of Pi anions driven by electrogenic H+ ejection.

Adenosine Diphosphate

[Oxidation of erucic acid and erucyl-CoA by isolated rat heart mitochondria: comparison to oleic acid].

The oxidation of [14 14-C] or [1 14-C] erucic acid by isolated mitochondria from Rat heart has been studied and compared with that of [10 14-C] oleic acid in varying conditions of incubation. Erucic acid is converted to CO2 and acid-soluble compounds much more slowly than oleic acid. The acid-soluble compounds which have been identified are acylcarnitines, ketone bodies and intermediates from the Krebs cycle; they are found in similar proportions for both substrates. Moreover, the oxidation rate of erucyl-CoA is comparable, if not equal, to that of oleyl-CoA in the same conditions. These results are discussed here. They lead to the conclusion that erucic acid is oxidized by isolated Rat heart mitochondria through the beta oxidation pathway, and that its oxidation is limited owing to its slow activation rate.

Animals

[Effect of hydrocortisone on the ribonucleic acid biosynthesis in the liver mitochondria, heart and spleen of rats with alloxan diabetes].

Mitochondrial RNA biosynthesis in diabetic rats under the influence of hydrocortisone (0.5 or 2 U per 100 g for 7 days) increased in the liver and decreased in the spleen. Heart mitochondrial RNA biosynthesis increased after hydrocortisone injection in a dose of 0.5 U per 100 g, but decreased after hormone administration in a dose of 2.5 U per 100 g. Thus, hydrocortisone played an important role in the changes of RNA biosynthesis in the liver, heart, and spleen mitochondria of diabetic animals.

Animals

Modulation of Ca2+ efflux from heart mitochondria.

The efflux of Ca2+ from rat heart mitochondria has been examined by using Ruthenium Red to inhibit active uptake after predetermined loadings with Ca2+. The efflux is proportional to the internal Ca2+ load; it is increased by Na+ applied when the mitochondria are respiring and this effect is inhibited by oligomycin. The efflux of Ca2+ is diminished by ATP and by ADP, with the latter the more effective. Both active uptake and efflux of Ca2+ are slowed by bongkrekic acid; this action has a time lag. The lower efflux found with the nucleotides and with bongkrekic acid seems to correspond to the more condensed state seen in the electron microscope when these agents are applied [Stoner & Sirak (1973) J. Cell Biol. 56, 51-64, 65-73]. The results are discussed in relation to the less-permeable state being contingent upon nucleotide binding to the membrane.

Adenine Nucleotides

[Anion-sensitive ATPase of the rat heart mitochondria].

The properties of anion-sensitive ATPase of rat heart mitochondria were studied. Na2CO3, NaHCO3 and Na2SO3 stimualted ATPase activity by 69, 41 and 110%, respectively. Azide, tiocinate and perchlorate inhibited bicarbonate-stimulated ATPase. Bivalent cations increased ATPase activity in such a sequence: Zn2+ greater than or equal to Cd2+ greater than or equal to Co2+ greater than or equal to Mg2+ greater than or equal to Mn2+ greater than Ni2+. In the presence of bicarbonate and sulfite. ATPase activity was maximally stimulated with magnesium. Ni2+ and Ca2+-ions inhibited Mg2+-dependent activity of bicarbonate-stimulated ATPase. AMP uninhibited ATPase activity. The 4 mM concentration of ADP inhibited activity of HCO-3-ATPase. Activity of ATPases decreased at lower temperatures. The properties of anion-sensitive ATPase of rat heart mitochondria and that of HCO-2-ATPase of other cells are discussed.

Adenine Nucleotides

Acute effects of acetaldehyde and ethanol on rat heart mitochondria.

The effects of acute exposure of rat heart mitochondria to ethanol (87, 65, and 45 mM) and acetaldehyde (3, 1, and 0.3 mM) were studied using both glutamate and pyruvate/malate substrates. Mitochondria assayed with pyruvate/malate substrate showed no apparent effects of acute exposure to or 30 min preincubation with ethanol at the three concentrations tested. With glutamate substrate, acute ethanol at 87 and 65 mM produced significant decreases in respiratory control ratio (RCR). With 65 and 45 mM acute ethanol, mitochondrial oxygen consumption (QO2) was significantly decreased. Acute acetaldehyde significantly decreased RCR and QO2 of mitochondria tested with both substrates. The depressive effect was more pronounced with pyruvate/malate substrate than with glutamate substrate. With pyruvate/malate substrate, the ADP/O ratio was also decreased with 3 and 1 mM acetaldehyde. Preincubation with acetaldehyde had no effect on mitochondrial function except for significantly decreased RCR after preincubation with 3 mM acetaldehyde followed by assay with glutamate substrate. The data indicate a depressant effect of acetaldehyde and ethanol on cardiac mitochondria that may contribute to abnormal cardiac biochemistry and function characteristic of alcoholic cardiomyopathy.

Acetaldehyde

Influence of Mg2+-ions on the properties of rat heart mitochondria in dependence on the preparation.

1. The preparation of rat heart mitochondria with Potter-Elvehjem homogenizer results in mitochondria showing stimualtion of respiration induced by Mg2+. This stimualtion is neither caused by adherent hexokinase nor by energy-dependent magnesium accumulation (Mg2+ content in the presence of 10 mM glutamate: 22 nmoles/mg protein; in the presence of glutamate plus antimycin A 21 nmoles/mg protein). 2. The effect of added magnesium is excluded by addition of carboxyatractyloside. This demonstrates the activity of an ATPase outside of the mitochondrial inner membrane. 3. A simple and rapid method for the preparation of Mg2+-insensitive rat heart mitochondria is presented. The minced heart is pressed through a normal syringe and then treated with trypsin. 4. A comparison of mitochondria of both preparations shows that there is no difference in magnesium content and no energy-dependent magnesium influx.

Adenylate Kinase