Hemocyanin in Oniscus asellus (Isopoda)

• 1.1. An immunological crossreactivity, as demonstrated by Western blotting, exists between O. asellus hemocyanin and anti-keyhole limpet (Megathura crenulata) hemocyanin.
• 2.2. Using the anti-keyhole limpet hemocyanin, the presence of hemocyanin was detected in the hepatopancreas.
• 3.3. The amino acid composition of the hemocyanin of O. asellus was found to be similar to that of other isopods.

     

Biochemical and functional characterization of Rapana thomasiana hemocyanin

• 1.1. The hemocyanin (Hc) of the marine gastropod mollusc Rapana thomasiana was collected from animals living on the west coast of the Black Sea and characterized for its biochemical and functional properties.
• 2.2. This Hc is very similar to other gastropod Hcs as far as amino acid composition, general structure and reactivity of the binuclear copper active site are concerned.
• 3.3. Some peculiarities in the dissociation-reassociation pattern are observed in comparison to other gastropod Hcs, in particular with respect to the ability to form sopramolecular aggregates.
• 4.4. Changes in pH disclose a strong reverse Bohr effect. Different R and T states are required to describe the oxygen binding curves at the different pHs.

     

Functional properties of the two major hemoglobin components from Leporinus friderici (Pisces)

• 1.1. The hemoglobins of Leporinus friderici were separated by liquid chromatography on DEAE-Sepharose in order to isolate the two major electrophoretic components.
• 2.2. The chromatographic fraction I (electrophoretically slow anodic) showed no Bohr effect and no nucleoside triphosphate modulation.
• 3.3. The chromatographic fraction III (electrophoretically fast anodic) showed a normal Bohr effect and addition of nucleoside triphosphate decreased oxygen affinity but did not alter the Bohr effect.
• 4.4. The whole hemolysate showed a normal Bohr effect and phosphate modulation altered both Bohr effect and oxygen affinity.
• 5.5. No or little difference between the effect of adenosine or guanosine triphosphates on hemoglobin function was observed.

     

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.

     

A dopamine- and octopamine-sensitive adenylate cyclase in the nervous system of Octopus vulgaris

• 1.1. Adenylate cyclase activity was assayed in the optic lobe of Octopus vulgaris.
• 2.2. Both octopamine and dopamine stimulate the octopus adenylate cyclase, apparently by competing with the same receptor site.
• 3.3. (±)-2-Amino-6,7-dihydroxy-1,2,3,4-tetrahydronaphthalene-HBr (6,7-ADTN) and a number of phenylethanolamine derivatives stimulate the octopus adenylate cyclase activity.
• 4.4. The dopamine D-1 antagonists R(+)-7-chloro-8-hydroxy-3-methyl-1-phenyl-2,3,4,5-tetrahydro-1H-3-benzazepine-HCl (SCH-23390) and (±)-7-bromo-8-hydroxy-3-methyl-1-phenyl-2,3,4,5-tetrahydro-1H-3-benzazepine-HCl (SKF-83566) are unable to antagonize the effects of dopamine and octopamine, and similarly ineffective is the agonist (±)-1-phenyl-2,3,4,5-tetrahydro-1H-3-benzazepine-7,8-diol-HCl (SKF-38393).
• 5.5. No detectable binding of labelled SCH-23390 occurs on membrane preparations from octopus optic lobe.

     

Seasonal adaptations in oxygen transport in brown trout Salmo trutta fario

• 1.1. Blood parameters determining oxygen capacity and oxygen affinity were measured in brown trout at different times of the year.
• 2.2. Haematological data indicate a slight decrease in blood oxygen capacity during the warm seasons. 3. Oxygen affinity increases significantly during summer and decreases in winter.
• 3.4. The changes in P₅₀ exhibited a positive correlation with the amount of anodic haemoglobin components, and a negative correlation with the amount of cathodic haemoglobin components.
• 4.5. The changes observed in the [ATP]/[Hb] molar ratio were not correlated with oxygen affinity and gave values near one.
• 5.6. We conclude that the oxygen affinity increase could be a physiological adaptation to oxygen transport during the wanner period. A possible mechanism is discussed.

     

A kinetic study of inosine nucleosidase from Azotobacter vinelandii

• 1.1. Inhibition of inosine nucleosidase from Azotobacter vinelandii by ATP and bases can be qualitatively and quantitatively accounted for by the partial noncompetitive inhibition mechanism with ligand exclusion model.
• 2.2. The enzyme has two binding sites for the substrate with equal affinity in the absence of the inhibitor. and two species of the inhibitor sites: I₁- and I₂-sites. The I₁-site may overlap part of each substrate binding sites, and the I₂-site is separated from the substrate sites.
• 3.3. ATP binds to the I₁-site of the enzyme, and prevents the substrate from binding to either of two identical sites, producing the cooperativity with inosine, whereas binding of ATP to the I₂-site causes a noncompetitive inhibition.
• 4.4. Adenine and hypoxanthine bind to the I₂-site of the enzyme, and the EIS complex is partially active, resulting in a partial noncompetitive inhibition with Michaelis-Menten kinetics.

     

Lactoferrin binding to human platelets

• 1.1. Platelets bind specifically lactoferrin.
• 2.2. The lactoferrin binding to the platelets depends on the concentration of labelled lactoferrin, the number of platelets, the time of incubation and pH.
• 3.3. The binding was characterized by two types of binding site: one with high affinity and low capacity, and another with low affinity and high capacity (respectively k_{aff 1} = 13.6 × 10⁹1/mol and about 40 binding sites, and K_{aff 2} = 1.23 × 10⁹1/mol and about 135 binding sites per platelet).
• 4.4. Both human transferrin and bovine lactoferrin compete with human lactoferrin for the receptors.
• 5.5. The presence of lactoferrin receptors on the platelet membrane surface is connected most probably with the effect(s) on the cell function(s) of these cells.

     

Sex-specific androgen binding in spotted hyaena (Crocuta crocuta) plasma

• 1.1. A charcoal adsorption assay demonstrated a large variance in androgen binding ability in female spotted hyaenas.
• 2.2. A positive correlation between plasma androgen binding ability and ovarian steroid concentrations was demonstrated in adult females.
• 3.3. The strong plasma binding affinity for testosterone and dihydrotestosterone (DHT) (nM) together with the lack of cortisol and weaker oestradiol-17β binding suggests that a specific androgen binding substance, possibly a protein, is present in adult females of this species.
• 4.4. The lack of high affinity binding in male spotted hyaenas is unusual and deserves further investigation.
• 5.5. Some androgen binding in all, including males and immature animals suggests that albumin may bind some plasma androgens in this species.

     

Effect of bovine milk antigens and egg lysozyme on the binding of 59Fe-lactoferrin to platelet plasma membranes

• 1.1. Platelets bind specifically to lactoferrin. A significant similarity between human lactoferrin and some bovine milk proteins has been established.
• 2.2. Because of the structural homology of lactoferrin and cows milk proteins they are able to influence lactoferrins regulatory function on the level of its binding to membrane receptors on platelets.
• 3.3. An inhibitory effect of bovine α-lactalbumin and of β-lactoglobulin on lactoferrin-receptor interaction was shown.
• 4.4. Bovine α-lactalbumin competes with lactoferrin for the binding sites.
• 5.5. Scatchard plot analysis of data shows one binding site for lactoferrin in the presence of α-lactalbumin with an affinity constant, Ka = 0.46 × 10⁹ mol/1 and 335 receptors/cell.
• 6.6. The inhibitory effect of β-lactoglobulin reaches 62% and is different for the common fraction ⨿-lactoglobulin and the genetic variants β-lactoglobulin A and B.
• 7.7. β-lactoglobulin does not compete with lactoferrin for the membrane receptors.
• 8.8. Bovine casein and egg lysozyme stimulate ⁵⁹Fe-lactoferrin binding to the receptors. The mechanism of these effects is still unknown.
• 9.9. Tested alimentary antigens are able to interact with lactoferrin and also with some platelet membrane structures.
• 10.10. Established changes in lactoferrin binding to the platelet membrane might be in relation to lactoferrins regulatory function and (or) eliminating mechanisms of these alimentary antigens.

     

Variations in response to environmental hypoxia of different colour forms of the shore crab,Carcinus maenas

• 1.1. The oxygen consumption of red and green Carcinus in normoxic and hypoxic sea water was determined, using an oxygen electrode in a sealed respirometer.
• 2.2. The red crabs had significantly higher “excited” oxygen uptake rates and a lower ability to compensate for hypoxia than the green crabs.
• 3.3. Red Carcinus display an emersion response to declining oxygen at lower oxygen tensions than the green crabs.
• 4.4. Mortality of red crabs exposed to prolonged anoxia was much greater.
• 5.5. The relationship of these findings to the zonation of the two colour forms on the shore is discussed.

     

Soluble and immobilized RP. palustris rhodanese—II. Some particular kinetic behaviour

• 1.1. Kinetic studies were carried out on the soluble and immobilized Rhodanese.
• 2.2. The soluble enzyme showed a typical Michaelis-Menten behaviour, an inhibitory effect was observed at high thiosulphate and cyanide concentrations.
• 3.3. The product sulphite was also an inhibitor, instead thiocyanate increased the enzyme velocity when it was added to the incubation mixture.
• 4.4. A ping-pong mechanism was proposed for Rp. palustris Rhodanese with a stable (free enzyme: E) and an unstable (sulfur substituted enzyme: ES) kinetic enzyme form.
• 5.5. The insolubilized Rhodanese presented an unusual kinetic behaviour, with sigmoid shape substrate profiles and non-linear double reciprocal plots.
• 6.6. From the empirical Hill equation, positive cooperativity (n>1) was found for both substrates.

     

Purification and partial characterization of an AMP deaminase from the marine invertebrate Palaemon serratus

• 1.1. AMP deaminase from Palaemon serratus tail muscle was partially purified by chromatography on cellulose phosphate.
• 2.2. Muscle homogenates expressed very low enzyme activities and the presence of ATP was necessary to detect AMP deaminase. The specific activity and substrate affinity of the purified enzyme were also very low.
• 3.3. The purified prawn muscle AMP deaminase was contaminated by contractile proteins, one of the major contaminants being actin.
• 4.4. The enzyme displayed a very high affinity for actomyosin which was only partially abolished by pyrophosphate.

     

Comparative study of hemoglobins from different Artemia populations. The influence of temperature on the oxygen equilibria

• 1.1. The P50 values of extracellular hemoglobin (Hb) of five Artemia populations from different geographical origin are affected by temperature.
• 2.2. The free oxygen binding energy is high for all the populations (ΔH between −34.7 and −56.2kj/mol).
• 3.3. A possible correlation between thermal sensitivity of Hb and the ambient temperature of the habitat must be considered very carefully.
• 4.4. The occurence of different quantities of Hb1 (αα chains) Hb2 (αβ chains) and Hb3 (ββ chains) in the different populations possibly influences thermal sensitivity.

     

Characterization of diverse hemolymph lectins in the cotton caterpillar Spodoptera littoralis

• 1.1. Hemolymph lectins (agglutinins) of the cotton caterpillar Spodoptera littoralis were analyzed by agglutination, cross-absorption and carbohydrate-hemagglutination inhibition using several vertebrate erythrocytes.
• 2.2. Lectins were found to interact, with all tested erythrocytes, by binding to carbohydrate moieties but showing no definite specificity.
• 3.3. Disulphide bonds were probably absent as 2-ME treatment was ineffective.
• 4.4. By cross-absorption studies, we have proposed that the hemolymph contains multiple lectins.

     

Catalytic and inhibitory properties of a major molecular form of acetylcholinesterase isolated from Lygus hesperus knight (Hemiptera: Miridae)

• 1.1. The major form of acetylcholinesterase (AChE) from Lygus hesperus demonstrated a greater affinity to selected substrates than unresolved AChE.
• 2.2. The turnover numbers of the native AChE were 7000 min⁻¹ for acetylthiocoline, 4800 for acetyl-(β-methyl) thiocholine, 3000 for propionylthiocholine, and 390 for S-butyrylthiocholine.
• 3.3. Each molecule of the major form had two active sites and each subunit had one active site.
• 4.4. Paraoxon or dichlorvos had a higher affinity to the major AChE form than to the unresolved AChE, resulting in a higher potency for the inhibition.
• 5.5. Some references of comparison are also made with AChE from other animal species.

     

Monoclonal antibodies neutralizing the haemolytic activity of box jellyfish (Chironex fleckeri) tentacle extracts

• 1.1. Three monoclonal antibodies have been produced which neutralize in vitro the haemolytic activity present in tentacle extracts of the box jellyfish (Chironex fleckeri).
• 2.2. Two of these monoclonal antibodies bound specifically to a component of relative molecular mass 50,000 in tentacle extract on Western blots.
• 3.3. This binding only occurred when the extracts were electrophoresed under non-reducing conditions.
• 4.4. The third monoclonal antibody did not display binding to Western blots of tentacle extract under any of our experimental conditions.

     

Respiratory mechanisms and metabolic adaptations of an intertidal crab,Chasmagnathus granulata (Dana, 1851)

• 1.1. The ventilatory mechanism, gill area, sites of oxygen uptake, oxygen consumption and activity of a crab from south Brazil, Chasmagnathus granulata, were investigated.
• 2.2. The oxygen uptake seems to be restricted to the gill lamellae.
• 3.3. The gill area varies with the wet body weight, being relatively higher in smaller animals. There is not a significative reduction of the gill area in relation to species of the infralittoral zone.
• 4.4. C. granulata presents a mechanism for recirculating the water of its branchial chamber when exposed to atmospheric air.
• 5.5. The oxygen consumption and activity are reduced when the animals are exposed to atmospheric air. The reduction in the oxygen consumption may be related to the poorly adapted respiratory system, while the decrease in activity may be a mechanism for saving energy during this hypoxic period.

     

Laboratory studies on the oxygen consumption of the marine teleost,Lichia amia (Linnaeus, 1758)

• 1.1. The oxygen consumption of the marine teleost, Lichia amia was investigated under controlled laboratory conditions.
• 2.2. The routine oxygen consumption showed a strong circadian rhythm with the fish being mainly active during the light period.
• 3.3. The specific mass exponent (dimension: μg O₂/g/hr) is temperature independent and ranges from 0.27–0.29.
• 4.4. Starving the fish results in a mean decrease in active, routine and standard oxygen consumption of 21%, 24% and 20%, respectively.
• 5.5. Feecling led to an increase in the oxygen consumption of the teleosts, with the mean metabolic rate over the 24 hr that followed, being 58% and 50% higher for fish that had been starved for 162hr and 40 hr, respectively.
• 6.6. Apparent SDA showed some variation and ranged from 6.0 to 35.5%.
• 7.7. The results obtained are generally in agreement with those recorded for other teleosts.

     

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.