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Biomedical subjects

A A Boldyrev

Publications and source records attributed to A A Boldyrev.

At least 19 recordsLinked to original sources
PubMed1995

Characterization of the subunit isoforms of duck salt gland Na/K adenosine triphosphatase.

The N-terminal sequences of the alpha and beta subunits from the Na/K-ATPase of duck salt gland have been determined by automated Edman degradation chemistry. These sequences were compared to sequences previously reported for Na/K-ATPase subunits from other sources in order to determine the subunit isoform composition of the salt gland enzyme. The comparisons indicate that the duck salt gland enzyme is composed of an alpha-1 subunit and a beta-1 subunit. This subunit isoform composition is consistent with the involvement of this enzyme in sodium excretion as Na/K-ATPases in other tissues involved in sodium excretion also have this subunit isoform composition.

Amino Acid Sequence↗
PubMed1995

Inhibition of Na+/K(+)-ATPase by phenoxyl radicals of etoposide (VP-16): role of sulfhydryls oxidation.

In the present work, we studied the effects of phenoxyl radicals, generated by tyrosinase-catalyzed oxidation of a phenolic antitumor drug, Etoposide (VP-16), on a purified dog kidney Na+/K(+)-ATPase by characterizing interactions of VP-16 phenoxyl radicals with the enzyme's SH-groups by ESR and correlating the loss of the enzymatic activity with the oxidation of its SH-groups, and oxidation of VP-16. VP-16/tyrosinase caused inhibition of Na+/K(+)-ATPase which was dependent on the incubation time and concentration of tyrosinase. The inhibition of Na+/K(+)-ATPase was accompanied by a decrease of DTNB (5,5'-dithiobis-(2-nitrobenzoic acid)-titratable SH-groups. In the presence of Na+/K(+)-ATPase, a typical ESR signal of the VP-16 phenoxyl radical could be observed only following a lag period the duration of which was proportional to the concentration of the Na+/K(+)-ATPase added. Our HPLC measurements demonstrated that Na+/K(+)-ATPase protected VP-16 against tyrosinase-catalyzed oxidation. Combined these results suggest that redox-cycling of VP-16/VP-16 phenoxyl radical by SH-groups of Na+/K(+)-ATPase occurred. Ascorbate which is known to reduce the VP-16 phenoxyl radicals, protected the enzyme against inactivation, prevented oxidation of the enzyme's SH-groups. Reduction of VP-16 phenoxyl radicals by ascorbate was directly observed by the semidehydroascorbyl radical signal in the ESR spectra. VP-16 phenoxyl radical-induced oxidation of sulfhydryls and inhibition of the Na+/K(+)-ATPase may be responsible for at least some of its clinical side effects (e.g., cardiotoxicity) which can be prevented by ascorbate.

Animals↗
PubMed1995

A comparative study of Na+/K(+)-ATPases of duck salt gland and canine kidney: implications for the enzyme's reaction mechanism.

Highly purified preparations of duck salt gland and canine kidney Na+/K(+)-ATPases with comparable specific activities were used to clarify the causes of previously reported differences between the substrate-velocity curves of these enzymes. When assays were done under identical conditions (pH 7.4; 37 degrees C), and a wide range of closely spaced ATP concentrations were used, the curves of both enzymes exhibited intermediary plateaus, as noted before for the salt gland enzyme. The two enzymes also had the same numbers of phosphorylation and ouabain binding sites, and their catalytic subunits were of the alpha 1 isoform type as revealed by immunostaining with specific antibodies. The findings suggest that the substrate-velocity curves of all widely used Na+/K(+)-ATPases may contain an intermediary plateau which is diagnostic of reaction mechanisms that generate rate equations containing powers of substrate concentration greater than two, e.g., a mechanism involving an oligomer with more than two protomers.

Animals↗
PubMed1995

[Effect of ligands on rotational mobility of Na,K-ATPase].

Phosphorescence anisotropy of eosin-5'-isothiocyanate labelled Na,K-ATPase purified from duck salt glands has been studied. The initial anisotropy value is 0.235 +/- 0.015 (room temperature) and does not depend on the enzyme conformation (sodium or potassium). The experimental curve is fitted into a two-exponential curve with residual term, the fast component corresponds to the rotational mobility of the functional unit of Na,K-ATPase (promoter), while the slow one--to that of larger associates. In the presence of ligands modifying the conformational state of Na,K-ATPase (sodium, potassium, ATP) the rotational mobility of the fast component does not change in contrast with the slow one. A comparison of the enzyme rotational mobility in the presence of ligands simulating different steps of hydrolytic cycle suggests that interprotomer interactions are changed in the course of hydrolytic cycle: the fraction of larger associates increases at the step of the enzyme interactions with potassium ions, whereas their mobility in the bilayer enhances sharply after interaction with ATP. In the presence of the 2% non-ionic detergent, C12E9, the initial anisotropy value decreases down to 0.1; the residual term disappears thereby, while the curve is still two-exponential. However, the difference in the rotational mobility of sodium and potassium conformers diminishes. At the same time, the ratios between protomers and oligomers in the presence of sodium and potassium become approximated. This indicates that in the presence of the detergent high molecular weight associates are solubilized, the mobility of the both protomers and oligomers of Na,K-ATPase increases, while the difference between the mobilities of sodium and potassium conformers is disappeared.

Animals↗
PubMed1995

[Carnosine metabolism in excitable tissues: biological significance].

The paper provides the data characterizing the antioxidative activity of carnosine and its related compounds by using various complexity models wherein primarily various active oxygen forms are generated. Though the agent is able to interact with various radicals, it is more effective against a hydroxyl radical than homocarnosine, anserine, and acetylcarnosine. Among the compounds studied, acetylcarnosine and anserine provide the best protection against a superoxide anion, anserine is the most effective agent in suppressing the myeloperoxidase reaction. Homocarnosine is more effective than the other carnosine-related compounds, in protecting thymine from the damaging effect of ultraviolet irradiation. It is suggested that metabolism of these compounds makes various carnosine derivatives appear in relation to the tissue specificity of formation and conversion of active oxygen forms.

Animals↗
PubMed1992

[The effect of carnosine on Ca-channels in rabbit skeletal muscle sarcoplasmic reticulum].

Carnosine (beta-alanyl-L-histidine), which is present in millimolar concentrations in skeletal muscles, induces Ca2+ release from the heavy fraction of rabbit skeletal muscle sarcoplasmic reticulum by activation ruthenium red-sensitive Ca-release channels. The effect of carnosine is dose-dependent, which indicates the presence of saturable carnosine-binding sites in the Ca-release channel molecule. The half-maximal Ca2+ release is observed in the presence of 8.7 mM carnosine. At the same time, carnosine addition to the medium increases the affinity of sarcoplasmic reticulum Ca-channels for the Ca-release activators, caffeine and adenine nucleotides. It is concluded that carnosine is an endogenous regulator of skeletal muscle sarcoplasmic reticulum Ca-channels which modulates the affinity of these channels for different ligands.

Adenine Nucleotides↗
PubMed1992

[Characteristics of chloramine complexes of carnosine with hypochlorite anion].

Carnosine interaction with CIO results in the formation of a stable chloramine complex. The binding of the whole bulk of hypochlorite to carnosine is completed within one minute of incubation. During subsequent 2-hour incubation no more than 15% of the chloramine complex is destroyed; this property of carnosine makes it similar to taurine. Unlike histidine and beta-alanine, glutathione rapidly interacts with hypochlorite. However, in contrast with these compounds and carnosine, glutathione does not form stable chloramine complexes with CIO. The putative role of myeloperoxidase in the development of senile human lens opacities is discussed.

Animals↗
PubMed1992

[The effect of carnosine and trolox on cellular chemiluminescence of leukocytes].

The effects of hydrophilic antioxidant carnosine, trolox (6-hydroxy-2.5.7.8-tetramethylchroman-2-carboxylic acid), and superoxide dismutase on the myeloperoxidase activity of leukocytes, superoxide anion and active oxygen species generation have been studied. Physiological concentrations of carnosine have been shown to decrease the ability of human leukocytes to produce chemiluminescence as a result of myeloperoxidase activation. However, the chemiluminescence induced by the generation of the superoxide or its derivatives is unaffected by this process. Trolox does not inhibit the induction of superoxide-dependent chemiluminescence of leukocytes either.

Antioxidants↗
PubMed1992

[Direct measurement of the interaction of carnosine and its analogs with free radicals].

An ESR study of interactions of carnosine and its derivatives with free radicals has been carried out. In model systems the spin trap OH. radical adduct generation has been shown to decrease significantly in the presence of carnosine in a pronounced concentration-dependent manner. A comparative study of effects of some other histidine-containing dipeptides on this process has revealed a similarity in anserine, homocarnosine, and acetylcarnosine actions.

Animals↗
PubMed1992

[Carnosine: biological role and prospects for use in medicine].

The biological role of the histidine-containing dipeptide carnosine (beta-alanyl-L-histidine) has been reviewed. The properties and putative biological role of the dipeptide in vertebrate tissues are considered. The antioxidative activity of carnosine and related compounds is described. The author's conception of the membranoprotective effect of carnosine on cells, tissues, and whole organism has been formulated. The properties of carnosine as an antistressory radioprotective agent are discussed. The data presented suggest that carnosine is a perspective immunomodulating tool which has many applications in medicine.

Adjuvants, Immunologic↗
PubMed1992

[Carnosine and anserine in working muscles--study using proton NMR spectroscopy].

NMR spectroscopy was used to study carnosine and anserine metabolism in rat tissues under intensive muscle loading. Muscle loading was accompanied by the dipeptide (predominantly anserine) accumulation in muscle tissues. Preliminary per os administration of carnosine (250 mg/kg of body mass) did not increase the dipeptide content in muscle tissues but diminished the lactate content in rat muscles under intensive muscle loading.

Animals↗
PubMed1991

[Nonhyperbolic kinetics of Na,K-ATPase--a new viewpoint].

The kinetics of the 130 kDa monomer obtained by treatment of duck salt gland Na,K-ATPase with C12E8 was compared with that of the membrane-bound enzyme. The shapes of the substrate-velocity curves for the membrane-bound and solubilized forms were quite different: a hyperbolic one for the monomeric Na,K-ATPase and a nonhyperbolic one for the native enzyme. A reaction scheme for ATP hydrolysis based on a comparative analysis of kinetic properties of these two forms is proposed. Experimental evidence in favour of this hypothesis is presented.

Animals↗
PubMed1991

The mechanism of the modifying effect of ATP on Na(+)-K+ ATPase.

On the basis of a review of the literature and a study of the molecular and kinetic properties of Na(+)-K+ ATPase, a model is proposed that explains the regulation of the activity of the enzyme by ATP in terms of an acceleration of the E2----E1 transition. It is presumed that the transition occurs via a short-lived oligomer whose formation is accelerated by ATP. In the context of this model, the non-Michaelis-Menton kinetics of the enzyme can be explained by interprotomer interactions. After solubilization of the enzyme with octaethylene glycol dodecyl ether, the hydrolysis of ATP follows ordinary Michaelis-Menton kinetics. The validity of the model is also supported by radiation-inactivation experiments with a nucleotide (GTP) which does not accelerate the E2----E1 transition, as well as by experiments with a low concentration of ATP. In both situations, the size of the molecular target corresponds to the monomeric form of the enzyme.

Adenosine Triphosphate↗