Agaricus bisporus tyrosinase—II. Characterization of hydroxylase and dehydrogenase activities

• 1.1. Preparative Isoelectric focusing (PIEF) was used to isolate hydroxylasic and dehydrogenasic activities, at different pI.
• 2.2. The fraction at pI 4.7 and 4.9 displays a pure dehydrogenase activity (substrate l-DOPA).
• 3.3. This fraction did not react with tyrosine, either in the spot-test or in absorption spectra (200–620 nm), and did not exhibit any oxygen consumption.
• 4.4. The fraction at pI 4.1 and 4.3 reacted with both l-DOPA and tyrosine as substrate, showing dehydrogenase and hydroxylase activity.
• 5.5. The latter activity was confirmed by the oxygen consumption test, showing that molecular oxygen is used to ortho-hydroxylate tyrosine.

     

Prenatal induction of tyrosine aminotransferase in rat liver

• 1.1. Hepatic tyrosine aminotransferase activity from adult rat can be resolved into four components on hydroxylapatite column.
• 2.2. A similar profile of enzyme distribution can be obtained from late foetal liver.
• 3.3. Insulin administration to pregnant rats result in induction of two isoenzymes of tyrosine aminotransferase in foetal rat liver. Similarly Cyclic AMP injection to foetal rats in utero results in the induction of the same two forms of the enzyme.
• 4.4. Triaminolone injection to foetal rats in utero leads to the induction of three of the isoenzymes of tyrosine aminotransferase.

     

Tyrosine requirement in Tetrahymena revealed by nutritional amino acid imbalances

• 1.1. So far, tyrosine has been considered non-essential for growth and cell multiplication in Tetrahymena pyriformis and T. thermophila.
• 2.2. This report shows, however, that tyrosine becomes essential if phenylalanine becomes limiting.
• 3.3. The results indicate that two different transport systems are involved in uptake of tyrosine and phenylalanine.

     

Oxygen binding by the hemocyanin of Busycon canaliculatum

• 1.1. This study examined the effect of the monoamines dopamine and octopamine, as well as tyrosine on the oxygen affinity and cooperativity of oxygen binding by the hemocyanin of the marine gastropod Busycon canaliculatum. The effect of temperature on hemocyanin oxygen affinity was also examined.
• 2.2. Freezing Busycon hemocyanin did not affect the binding of oxygen.
• 3.3. Dopamine, octopamine and tyrosine had no significant effect on the oxygen affinity or cooperativity of oxygen binding by the hemocyanin of B. canaliculatum.
• 4.4. It was concluded that Busycon hemocyanin either has no binding sites for the two monoamines or for tyrosine, or that binding of the molecules has no functional significance.
• 5.5. Both temperature sensitivity and affinity of hemocyanin-oxygen binding were similar to values previously reported for hemocyanin of Busycon from other localities.

     

Interference of benzo[a]pyrene with cytochrome P-450-mediated steroid metabolism in pyloric caeca microsomes of the sea star Asterias rubens L.

• 1.1. The effects of benzo[a]pyrene (BaP) on the metabolism of progesterone and pregnenolone, and the effects of steroids on BaP metabolism were examined in pyloric caeca microsomes of female Asterias rubens.
• 2.2. The patterns of metabolism of progesterone and pregnenolone in microsomes were similar to those found in previous studies for homogenates and tissue incubations of pyloric caeca.
• 3.3. BaP reduced the rate of hydroxylation of progesterone and pregnenolone, but had no effect on metabolite formation by non-cytochrome P-450-catalysed reactions.
• 4.4. Microsomal BaP hydroxylase activity was reduced by the presence of progesterone, but pregnenolone and testosterone had no such effect.
• 5.5. The reductions in steroid or BaP metabolism were progressive with increasing ratios of the concentration of the interfering compound to that of the assay substrate and were maximally 50% or less at ratios of × 100.
• 6.6. It is concluded that isoenzymic forms of cytochrome P-450 are present, with preferences towards either steroid or BaP metabolism. The implications of the results for the in vivo situation are discussed.

     

Metabolism of testosterone in human lung in vitro

• 1.1. The metabolism of 4-¹⁴C-testosterone in human lung in vitro was investigated.
• 2.2. The metabolism was most pronounced in incubations of homogenated tissue, whereas it was rather restricted in the mitochondrial, microsomal and soluble fraction incubations.
• 3.3. The by far most prominent metabolite in all experiments was androst-4-ene-3,17-dione.
• 4.4. No slfate or glucuronide conjugation took place.

     

Lipoxygenase of the thermophilic bacteria thermoactinomyces vulgaris—properties and study on the active site

• 1.1. A lipoxygenase activity was purified from Thermoactinomyces vulgaris and some of its properties were characterized.
• 2.2. The enzyme showed a temperature activity range of 40–55°C with still significant activity over 60°C.
• 3.3. The pH of activity on linoleic acid had a broad range with an optimum at pH 6.0 and a weaker one at pH 11.0.
• 4.4. On arachidonic acid the pattern was narrow bell-shaped with an optimum at pH 6.5.
• 5.5. The purified lipoxygenase from Th. vulgaris showed an apparent K_m of 1 mM and V_max of 0.84 μmol diene/min/mg protein.
• 6.6. It was inhibited by the oxidation products, 9-HPOD and 13-HPOD.
• 7.7. A 160,000 Da molecular weight of the enzyme was determined by molecular filtration. Methionine, tyrosine, tryptophan and cysteine are apparently involved in its activity.

     

Terrestrial dormancy in the turtle Kinosternon flavescens: Respiratory metabolism and dehydration

Experimentally induced terrestrial dormancy resulted in reduced rates of oxygen consumption and carbon dioxide release by the turtle, Kinosternon flavescens.

• 1.2. An unusually low respiratory quotient for dormant turtles indicates retention of carbon dioxide.
• 2.3. Sharply elevated levels of oxygen consumption upon emergence from dormancy suggest payment of an oxygen debt and support previous suspicion of increased anaerobic metabolism.
• 3.4. Most of the weight lost by dormant turtles is attributed to dehydration rather than metabolism.
• 4.5. Suppressed gas exchange apparently reduces water loss from the respiratory tract, an adjustment which may be essential in avoiding severe dehydration.

     

Changes in high-energy phosphate metabolism in the water scorpion,Ranatra chinensis,under cold water-warm water stress

• 1.1. To evaluate changes in high-energy phosphate metabolism in the water scorpion (Ranatra chinensis) under restraint and cold water-warm water stresses, in vivo [³¹P]NMR spectra were obtained.
• 2.2. Under restraint stress, arginine phosphate (Arg-P) decreased by 10% after 1 hr and remained at that level thereafter, while β-ATP showed negligible changes over 6 hr.
• 3.3. As the water temperature gradually increased or decreased, the relative concentration of Arg-P decreased due to enzyme regulation.
• 4.4. Repeated cold water-warm water stress, which consisted of repeated 15 min exposures to cold water (5°C) followed by 15 min exposures to warm water (30°C) caused distinct decreases in Arg-P and β-ATP concentration. These decreases were dependent on the frequency of exposure.
• 5.5. Phosphomonoesters (PME) increased not only with restraint stress but also with cold water-warm water stress.
• 6.6. The effect of cold water-warm water stress on high-energy phosphate metabolism was greater than that of restraint stress.

     

Metabolism of ingested indole compounds in the gut of three species of heteroptera

• 1.1. All kinds of indole compounds used for the experiment were more or less metabolized in the gut of Dolycoris baccarum, Eurydema rugosum and Elasmostethus humeralis.
• 2.2. The metabolic pattern of the bugs was classified into four types (I–IV) for several indole compounds in the same way as for IAA.
• 3.3. The IAA metabolites in the excreta of the three species were probably the high-molecular compound combining with such substances as amino acids, sugars or proteins to some position of indole nucleus.
• 4.4. The crude excreta of E. humeralis fed with each of several indole compounds had a significant auxin activity.
• 5.5. Most of the metabolites of indole-3-acetaldehyde in the excreta of E. humeralis had also a significant auxin activity.
• 6.6. The significance of auxin metabolism in the gut of the bugs and the difference of auxin metabolism between aphids and bugs are discussed.

     

Effect of conjugation on the incorporation of glycine into Tetrahymena

• 1.1. Conjugation of Tetrahymena enhanced the incorporation of glycine into the nuclear fraction by 500%.
• 2.2. Incorporation of glycine into the microsomal supernatant was augmented by almost 500% by conjugation.
• 3.3. Mitochondrial incorporation was stimulated nearly 3-fold in the conjugating strains while the incorporation of glycine into the microsomes was enhanced approximately 2.5 times.
• 4.4. In the whole cell, glycine incorporation was increased nearly 2-fold by conjugation.
• 5.5. Strong nuclear involvement was indicated by elevated metabolic activity and incorporation of glycine into RNA and DNA.
• 6.6. Stimulation of the metabolism of Tetrahymena by cell communication suggests that the contents of a cell can have a synergistic effect on another cell.
• 7.7. Augmentation of the biosynthetic capacities of cells by fusion is a demonstration of the dominant role of the cell membrane in the regulation and control of cells.
• 8.8. Enhancement of biosynthesis of nuclear proteins in conjugating strains of cells indicates that fusion gives rise to the synthesis of new protein from previously existing protein or protein procursors.
• 9.9. The specific activities of the subcellular fractions after the incorporation of glycine into 2 separated starved strains of Tetrahymena followed the usual pattern of nucleus less than whole cells, whole cells less than mitochondria, mitochondria less than microsomes, but with the microsomal supernatant being much greater than that of the microsomes.

     

The influence of anaerobiosis on the levels of adenosine nucleotides and some glycolytic metabolites in tubifex sp. (annelida,oligochaeta)

• 1.1. In Tubifex sp. the amounts of ATP, ADP and AMP, and of glucose, glucose-1-P, glucose-1-P, glucose-6-P, fructose-6-P and fructose-1, 6-P were measured after experimental anaerobiosis.
• 2.2. The energy charge decreased from 0.84 to 0.07/0.69 within 6–9 hr of anaerobiosis.
• 3.3. During long term anaerobiosis there was no change from 0.70/0.69.
• 4.4. The concentrations of glucose, glucose-6-P and fructose-1,6-P increased somewhat during an initial phase of anaerobiosis.
• 5.5. The data are discussed with respect to the regulation of energy metabolism, especially during the transition of aerobic to anaerobic metabolism.
• 6.6. It is concluded that this transition is accomplished within 6–12 hours.

     

Temperature regulation and metabolic rhythms in populations of the house sparrow,Passer domesticus

• 1.1. The house sparrow, Passer domesticus, has a circadian rhthym of metabolism and body temperature.
• 2.2. Evolutionary adaptation to a hot and humid climate is reflected in the lower metabolism and greater insulation of the Houston population than observed in populations from Ann Arbor, Michigan; Boulder, Colorado and Syracuse, New York.
• 3.3. There are no significant differences in either body temperature or evaporative water loss of all four populations.
• 4.4. The Houston population is able to survive higher ambient temperatures than is found in the Ann Arbor, Michigan or Boulder, Colorado population.

     

The energy costs of temperature regulation in birds: The influence of quick sinusoidal temperature fluctuations on the gaseous metabolism of the japanese quail (Coturnix coturnix japonica)

• 1.1. Quick sinusoidal temperature fluctuations (constant average 10°C) cause an increase in metabolism in comparison to an invariable constant ambient temperature of the same dimension.
• 2.2. At the observed mean value of 10°C metabolism is increased by 0.8% per 1 K/hr based on the values of resting metabolic rate (correlation: M = 53.5 + 0.445 T_a, M in J/K g hr, T_a = ambient temperature change in K/hr) and 0.6% based on the values of activity metabolism (M = 70.4 + 0.425 T_a).
• 3.3. The absolute augmentation of metabolism per 1 K/hr is, by comparison, the same for day and night. Its amount is 0.42 and 0.43 J/K g hr respectively.
• 4.4. In the response of metabolism to temperature fluctuations no differences could be found with respect to the amplitude and frequency modifications of temperature.
• 5.5. The increase of energy consumption is probably caused to a greater extent by “overshoot” of the feedback control system in the course of adjusting metabolism to new levels according to the ambient temperature conditions.
• 6.6. Short term ambient temperature changes (i.e. measuring different temperature levels in one night to test basic metabolism vs ambient temperature) cannot produce reasonable values for basic metabolic rate, since these artificially high values reflect the testing procedure.

     

Ventilatory rate in the butterfish (Pholis gunnellus) as a consequence of temperature and previous emersion

• 1.1. Observation of ventilation in immersed Pholis gunnellus showed a linear relationship between ventilatory rate and temperature between 8 and 20°C.
• 2.2. At 13°C and after 30 min emersion, ventilatory rate was initially lower than prior to emersion, providing evidence of adequate uptake of O₂ for standard metabolism during the emersion period.
• 3.3. This species has a laterally elongate body form with reduced scales and extensive mucus secretion.
• 4.4. During emersion, gaping behaviour probably exposes the gills and extensively vascularised oesophageal regions to air.
• 5.5. These are considered to be morphological and behavioural adaptations by P. gunnellus, to aerial respiration in the intertidal habitats occupied by this species.

     

Aerobic metabolism,octopine production and phosphoarginine as sources of energy in the phasic and catch adductor muscles of the giant scallop placopecten magellanicus during swimming and the subsequent recovery period

• 1.1. The energy contributions of aerobic metabolism, phosphoarginine, ATP and octopine in the adductor muscles of P. magellanicus were examined during swimming and recovery.
• 2.2. A linear relationship was observed between the size of the phosphoarginine pool and the number of valve snaps. A linear increase in arginine occurred during the same period.
• 3.3. Octopine was formed during the first few hours of recovery, particularly in the phasic muscle.
• 4.4. The restoration of the phosphoarginine pool appeared to be by aerobic metabolism.
• 5.5. It is concluded that the role of octopine formation is to supply energy when the tissues are anoxic and to operate at such a rate as to maintain the basal rate of energy production.

     

Characterization of Colorado potato beetle lipophorin: A hydrocarbon-rich diacylgycerol-poor lipophorin

• 1.1. The lipid composition of lipophorin from the Colorado potato beetle, Leptinotarsa decemlineata Say, was analyzed.
• 2.2. This insect lipophorin contains 44% lipid and is characterized by large amounts of hydrocarbons and small amounts of diacylglycerol.
• 3.3. This is the first observation of a diacylglycerol-poor insect lipophorin in haemolymph.
• 4.4. Since the main energy source for flight in the Colorado potato beetle is proline, the low diacylglycerol content in lipophorin must be related to its peculiar flight metabolism.
• 5.5. This lipophorin, however, can still take up appreciable amounts of diacylglycerol from the locust fat body. Hydrocarbon uptake by this lipophorin was also demonstrated.
• 6.6. The main function of this lipophorin therefore seems to be transport of hydrocarbons from oenocytes to the cuticle.

     

Effects of the nonsteroidal anti-inflammatory drug piroxicam on energy metabolism in the perfused rat liver

• 1.1. The actions of piroxicam, a nonsteroidal and noncarboxylic anti-inflammatory drug, on the metabolism of the isolated perfused rat liver were investigated. The main purpose was to verify if piroxicam is also active on glycogenolysis and energy metabolism, as demonstrated for several carboxylic nonsteroidal anti-inflammatories.
• 2.2. Piroxicam increased oxygen consumption in livers from both fed and fasted rats.
• 3.3. Piroxicam increased glucose release and glycolysis from endogenous glycogen (glycogenolysis).
• 4.4. Gluconeogenesis from lactate plus pyruvate was inhibited.
• 5.5. The action of piroxicam on oxygen consumption was blocked by antimycin A, but not by atractyloside.
• 6.6. The action of piroxicam in the perfused rat liver metabolism seems to be a consequence of its action on mitochondria.
• 7.7. It can be concluded that inhibition of energy metabolism and stimulation of glycogenolysis are not specific properties of carboxylic nonsteroidal anti-inflammatory drugs.

     

Organ specific changes in energy metabolism due to anaerobiosis in the sea mussel Mytilus edulis (L.)

• 1.1. Anaerobic energy metabolism was investigated in different organs of Mytilus edulis and the whole animal.
• 2.2. Succinate accumulates to high levels in most organs but remains low in the hemolymph.
• 3.3. After 16 hours propionate accumulation is observed in all organs. Experimental evidence is not sufficient yet to point out organs that produce more propionate than others.
• 4.4. Acetate is a minor end product.
• 5.5. Acetate and propionate are found in the hemolymph in amounts equal to those in the organs.
• 6.6. Animals incubated in oxygen-free seawater accumulate more end products than animals exposed to air, in the form of volatile fatty acids that are excreted into the incubation water.
• 7.7. Alanine and glutamine increase in the posterior adductor muscle. Aspartate decreases in the total animal, posterior adductor muscle and gills, while in the hemolymph decrease in alanine, asparagine, serine, threonine and proline are observed.

     

Essential tyrosyl residues of human placental alkaline phosphatase

• 1.1. Human placental alkaline phosphatase was inactivated with tetranitromethane in a biphasic process.
• 2.2. Spectral and amino acid analysis demonstrated that the inactivation was due to the conversion of tyrosine residues to 3-nitrotyrosine.
• 3.3. The inactivation process showed saturation kinetics.
• 4.4. Protection of the enzyme against tetranitromethane inactivation was afforded by inorganic phosphate.
• 5.5. The binding affinity between the modified enzyme and inorganic phosphate was decreased.
• 6.6. Our results suggest the involvement of tyrosyl residues in the locus of phosphoryl site of the phosphorylated enzyme forms.