Mitochondrial aldehyde reductase: identification and characterization in rat liver and kidney cortex.

Aldehyde reductase (EC 1.1.1.2) has been regarded so far as an exclusively cytosolic enzyme. The present investigation shows that mitochondria of rat liver, kidney cortex and, tentatively, heart also contain an enzyme catalyzing oxidation of NADPH by aldehydes, p-nitrobenzaldehyde, methylglyoxal and glyceraldehyde. Activity of the mitochondrial enzyme can only be measured after the organelles are disrupted by sonication or solubilized with nonionic detergents. Mitochondrial aldehyde reductase activity contributed to about 4.6% and 2.5% of the total cellular activity in liver and kidney cortex, respectively. However, the specific activity in liver mitochondria was about one third and in kidney cortex mitochondria one tenth of that in the cytosol of the corresponding organ. The mitochondrial enzyme resembled the cytosolic one by its absolute specificity towards NADPH as the electron donor, a similar profile of aldehydic electron acceptors and identical Km values. Mitochondrial aldehyde reductase differed from the cytosolic enzyme by low sensitivity to known inhibitors of cytosolic aldehyde reductase, AL-1576, AL-4114 and ONO-2235. In liver, about 60% of the mitochondrial activity was tightly bound to the membranes whereas about 40% was present in the mitochondrial matrix. The membrane-bound activity was inactivated by digestion of mitoplasts with trypsin, alpha-chymotrypsin or papain, thus pointing to exposition of the substrate-binding site at the external surface of the inner membrane. On the other hand, latency of the enzyme in intact mitochondria indicates that the NADPH-binding site is located at the inner surface. These data provide the first direct evidence for the existence of aldehyde reductase in mitochondria of some rat tissues.

[Effect of paracetamol on the microsomal oxidation, oxidative phosphorylation and liver mitochondria swelling in rats of various ages]. / Vliianie paratsetamola na mikrosomal'noe okislenie, okislitel'noe fosforilirovanie i nabukhanie mitokhondrii pecheni krys raznogo vozrasta.

The effect of hepatotoxic dose of paracetamol (800 mg per kg, intraperitoneally, once a day during two days) on the system of microsomal oxidation, respiration, oxidative phosphorylation and high amplitude swelling of liver mitochondria was studied on 1-, 4- and 30-months old Wistar male rats. It has been shown, that paracetamol injection leads to the decrease of content of cytochrome P-450, to disorders of the function of monooxygenase system (the aminopyrine-N-demethylase and aniline hydroxylase activities were diminished), mitochondria macrostructure (the mitochondria high amplitude swelling time was decreased) and function (the respiratory control was decreased). These alterations have been observed to manifest to more extent in the liver of young rats as compared with old ones.

[The effect of osmotic pressure on oligomycin-sensitive ATPase activity and the liberation of Mg2+ from liver mitochondria of rats of various ages]. / Vliianie osmoticheskogo davleniia na oligomitsinchuvstvitel'nuiu ATP- aznuiu aktivnost' i osvobozhdenie Mg2+ iz mitokhondrii pecheni krys raznogo vozrasta.

The influence of osmotic pressure of the incubating medium (25-500 mM sucrose) on oligomycin--sensitive, 2,4-dinitrophenyl-stimulated ATP-ase-activity, Mg2+ release and swelling of the liver mitochondria in 1-, 3-, 12-, 24-months Wistar rats is, investigated to determine age changes of structurally functional state of mitochondria. An increase in the sucrose concentration in the medium from 150 to 500 mM causes almost equal and practically absolute inhibition of ATP-ase-activity in different-age groups of rats, regardless of the presence or absence of Mg2+ ions in the medium A fall of the sucrose concentration to 150-25 mM induces a decrease in mitochondria ATP-ase-activity in Mg2+ free medium in 12- and 24-months rats (to 30 and 22%, respectively). No changes are observed in 1- and 3-months animals. Differences in rates of exogenous NADH oxidation by mitochondria of 1- and 12-months rats as a reflection of inner membrane damage degree are not observed under these conditions. Relative changes in ATP-ase-activity in a Mg2+ free medium with sucrose concentration of 25 mM (compared with 150 mM) correlate (r = 0.82) with those of optical density of mitochondria, measured at light wave length of 520 nm. It is obvious that the liver mitochondria of young and old rats sufficiently differ in spontaneous swelling rate in the media with different osmotic pressure: mitochondria of 1-month rats swell much faster than those of old rats. Considerable age differences of osmotic dependence of Mg2+ output from mitochondria are observed. They depend also on peculiarities of spontaneous organelle swelling dynamics.(ABSTRACT TRUNCATED AT 250 WORDS)

[Age-related differences in the effect of osmotic pressure on liver mitochondrial respiration in rats]. / Vozrastnye razlichiia vliianiia osmoticheskogo davleniia na dykhanie mitokhondrii pecheni krys.

The subject under investigation is the influence of osmotic pressure of incubation medium (25-500 mM of sucrose) upon the respiration and the respiration control (RC) of mitochondria of the liver of rats aged 1, 3, 12 and 24 months when oxidizing succinate. In a medium with 0.3 M of sucrose the respiration rate under condition 3 (V3) and RC increased from the age of 1 to 12 months and decreased by 24 months. In a medium with 0.15 M of sucrose the age differences have not been observed. In a uncoupling state the osmotic dependence of the respiration of mitochondria of 1- and 12-month-old rats did not vary. It is assumed that with age there is a change in the rate of structural coupling of the carrier of adenine nucleotides with H(+)-ATP synthetase complex and (or) the viscosity of the matrix.