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4.
Biofizika ; 29(4): 640-2, 1984.
Article in Russian | MEDLINE | ID: mdl-6091786

ABSTRACT

Below 37 degrees the mobility of lipids in rat brain synaptic membranes evaluated by the spin probe method is higher in old (24-26 months) than in young (5-6 months) animals. The number of ouabain receptors in membranes increases with age, while maximum rate and Km values are not changed. The relation between structural state of membrane and properties of ouabain binding centers is discussed.


Subject(s)
Aging , Brain/metabolism , Synaptic Membranes/metabolism , Animals , Binding Sites , Brain/enzymology , Cell Membrane/metabolism , Lipid Bilayers/metabolism , Rats , Receptors, Drug/metabolism , Sodium-Potassium-Exchanging ATPase/metabolism , Spin Labels , Synaptic Membranes/enzymology , Temperature
5.
Biofizika ; 28(4): 709-11, 1983.
Article in Russian | MEDLINE | ID: mdl-6615913

ABSTRACT

In old rats in contrast to young and mature ones the treatment of synaptic membranes with SH-reagent N-ethylmaleimide, cholinergic agonist carbamylcholine or both of them decreases the specific binding of labeled muscarinic antagonist 3H-quinuclidinylbenzylate. The age related change of receptor conformation with exposure of thiol groups responsible for antagonist binding is suggested.


Subject(s)
Brain/growth & development , Receptors, Muscarinic/metabolism , Synaptic Membranes/metabolism , Aging , Animals , Brain/metabolism , Ethylmaleimide/pharmacology , Kinetics , Quinuclidinyl Benzilate/metabolism , Rats
6.
Prikl Biokhim Mikrobiol ; 19(2): 209-16, 1983.
Article in Russian | MEDLINE | ID: mdl-6856591

ABSTRACT

Luciferase of fireflies Luciola mingrelica was immobilized on cellulose films activated by cyanuric chloride or sodium periodate. Kinetic properties and the contribution of diffusional obstacles to the kinetics of the immobilized enzyme were examined. External and internal diffusion were found to influence the kinetic parameters. The stability of the enzyme was investigated at 25 degrees C and pH 7.8. Thermoactivation of the immobilized enzyme was shown to proceed in two stages: fast and slow. Dithiotreitol and cystein stabilized the enzyme at the fast stage while salt supplements at both stages. The fast thermoinactivation stage was apparently associated with the oxidation of luciferase SH-groups. It is demonstrated that the immobilized enzyme of Luciola mingrelica can be employed to measure ATP traces with the detection limit 0.1 mM. The enzyme immobilized on cellulose films can be used repeatedly.


Subject(s)
Cellulose/pharmacology , Coleoptera/enzymology , Enzymes, Immobilized/pharmacology , Hot Temperature , Luciferases/pharmacology , Adenosine Triphosphate/analysis , Animals , Diffusion , Drug Stability , Hydrogen-Ion Concentration , Indicators and Reagents/pharmacology , Kinetics
7.
Biofizika ; 27(4): 650-2, 1982.
Article in Russian | MEDLINE | ID: mdl-6289920

ABSTRACT

In the brain synaptic membranes from old rats (24-26 months) in comparison with the mature ones (6-7 months) an increase was shown in disintegration rate with DDS Na, mobility of 5-, 16-doxyl-stearate spin probes and in thermosensitivity of acetylcholinesterase. The total number of SH-groups decreased. While protein composition remained constant lysophosphatidylcholine and saturated fatty acid content in old animals increased. The age-dependent structural rearrangement of synaptic membranes is suggested.


Subject(s)
Brain/growth & development , Synaptic Membranes/physiology , Aging , Animals , Electron Spin Resonance Spectroscopy , Rats , Sodium Dodecyl Sulfate , Temperature
8.
Biokhimiia ; 47(5): 760-6, 1982 May.
Article in Russian | MEDLINE | ID: mdl-7093378

ABSTRACT

It was shown that the dimers of the firefly luciferase possess the catalytic activity, whereas the monomers do not. The dissociation constant (Kd) for active dimers was determined at pH 7.0--8.4 within the temperature range of 15--35 degrees and at MgSO4 and Na2SO4 concentrations varying from 37 to 370 mM and 49 to 490 mM, respectively. Under variable conditions the Kd value changed only insignificantly and made up to 13 nm. The substitution of Na2SO4 for MgSO4 decreased Kd 2.5 times. The effective rate constant for the enzyme inactivation (kin) was increased more than 5-fold, when the luciferase concentration was decreased from 200 down to 3.5 nM in the presence of 37 mM MgSO4. When the concentration of the latter was increased up to 185 mM, the value of kin ceased to depend on the enzyme concentration. The decrease of kin was also observed at an increase in Na2SO4. An inactivation pattern for the enzyme in solution was determined both for the monomer and for the dimer of the enzyme. The equations allowing to calculate the inactivation constant for the monomer (Ki) and dimer (k2) at different pH values, temperatures and salt concentrations were obtained. The enzyme was found to be stabilized by salts more than 10-fold, the stabilizing effect being far more pronounced for the enzyme monomer than for the dimer. The dependence of the effective kin value on pH and temperature was primarily influenced by the dependence of the inactivation rate constant for the dimer.


Subject(s)
Coleoptera/enzymology , Luciferases/metabolism , Animals , Hot Temperature , Kinetics , Macromolecular Substances , Magnesium/pharmacology , Osmolar Concentration , Sodium/pharmacology
9.
Biofizika ; 27(1): 156-8, 1982.
Article in Russian | MEDLINE | ID: mdl-6279169

ABSTRACT

Affinity of beta-adrenoreceptors in the rat brain synaptic membranes to agonists isoproterenol and norepinephrine, as well as to antagonist 125I-hydroxybenzylpindolol is lower in young (1 month) and old (24--26 months) than in mature (8--12 months) rats. Desensitization toward isoproterenol is expressed in the young ones only. In the old but not in other groups simultaneous action of isoproterenol and N-ethylmaleimide decreases the following binding of the antagonist while the same agents added separately produced no effect. It is suggested that beta-adrenoreceptors undergo age-related changes in their conformational state due to modification of the membrane environment.


Subject(s)
Brain/growth & development , Receptors, Adrenergic, beta/metabolism , Receptors, Adrenergic/metabolism , Synaptic Membranes/metabolism , Aging , Animals , Benzyl Alcohols/metabolism , Brain/metabolism , Isoproterenol/metabolism , Norepinephrine/metabolism , Pindolol/analogs & derivatives , Pindolol/metabolism , Rats , Substrate Specificity
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