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Biomedical subjects

A A Akhrem

Publications and source records attributed to A A Akhrem.

At least 109 records · Page 6Linked to original sources
PubMed1979

Methylation analysis in glycoprotein chemistry. General procedure for quantification of the products of solvolysis of permethylated glycopeptides and glycoproteins.

A technique for the gas chromatographic analysis of the products of solvolysis of permethylated glycopeptides and glycoproteins has been developed. It involves methanolysis of a permethylated compound, quantitative transformation of methyl ethers of methyl glycosides into the corresponding O-trimethylsilyl(TMS)-O-methylalditols [2-deoxy-2-(N-methyl)acetamido-O-TMS-O-methylalditols in the case of hexosamine derivatives] and gas chromatographic quantification using a single column packed with 0.4% OV-225 on surface-modified Chromosorb.

Acetylglucosamine↗
PubMed1979

[Characterization of human retroplacental blood plasma proteins isolated by biospecific chromatography on immobilized cortisol].

Human retroplacental blood plasma proteins with affinity for cortisol were isolated by biospecific chromatography and identified by electrophoretic and immunochemical methods as alpha1- and beta1-globulins and IgG. IgM and IgA immunoglobulins. A high specific affinity for cortisol (Kas = 1,5 . 10(8) M-1 at 23 degrees C) and progesterone (Kas = 2,0 . 10(8) M-1 at 23 degrees C) was observed only for alpha-globulin; other proteins had a low affinity for cortisol. The molecular weight of alpha1-globulin (transcortin) was found to be 50,000-55,000. The amino acid and monosaccharide compositions of this glycoprotein were studied. Its N-terminal and C-terminal amino acid sequences are: Met-Asx-Pro-Asx-Ala- and (Val, Gln)-Leu, respectively. It was concluded that under normal physiological conditions and during pregnancy transcortin is the only specific corticosteroid-binding plasma protein. A complete removal of bound cortisol from the protein mixture and subsequent hydroxylapatite chromatography resulted in homogeneous transcortin retaining more than 90% of its binding capacity. The formation of the transcortin-steroid complex and its complete dissociation are accompanied by conformational changes of the protein globule. Significant changes of the spectral properties of the tryptophane residue of protein and the steroid delta4-3-keto group are indicative of the possibility of their direct interaction.

Amino Acid Sequence↗
PubMed1979

[Reversible precipitation of the glucocorticoid receptor from rat liver by La3+ ions].

It was shown that La3+ ions are capable of precipitating the glucocorticoid receptor from rat liver. Treatment of cytosol containing either free receptor or the hormonereceptor complex with 0,005 M La(NO3)3 results in recptor precipitation. The pellet is readily dissolved in buffer with EDTA. Transcortin and transcortin-like proteins are not affected by La3+ ions. The lanthanium-treated receptor does not lose its ability to bind DNA and chromatin in vitro. It is suggested that precipitation by La3+ ions can be used for separation of the receptor from transcortin and transcortin-like proteins as well as for evaluation of binding parameters for steroid-receptor complexes in rat liver cytosol.

Animals↗
PubMed1979

Isolation, structural organization and mechanism of action of mitochondrial steroid hydroxylating systems.

A scheme for the isolation of individual proteins of the 20S,22R-cholesterol hydroxylating system is presented. Physico-chemical and structural parameters of cytochrome P-450 are furnished. Self-reconstitution of indivudal proteins of the system into an enzyme unit is shown. Furthermore a regulation process of electron transport in the 20S,22R-cholesterol system is considered.

Adrenodoxin↗
PubMed1979

The mechanism of hydroperoxide-dependent reactions with participation of cytochrome P-450.

Cytochrome P-450 destruction kinetics by cumene hydroperoxide (CHP) has been studied at 25 degrees C in phosphate buffer, pH 7.25-7.50, in various systems: intact and induced rat or rabbit microsomes, highly purified LM2- and LM2- and LM4-forms of cytochrome P-450 from rabbit liver microsomes. The destruction kinetics is characterized by three phases in all systems. The CHP-influenced cytochrome P-450 destruction is a radical chain process with linear termination of the chains. The acidic phospholipids, phosphatidylserine and phosphatidylinositol and total microsomal phospholipids containing the acidic lipid components activate cytochrome P-450 in the hydroxylation of aniline and naphthalene by CHP. Phosphatidylcholine and sphingomyelin have no effect upon the cytochrome P-450 activity in the type I and II substrates oxidation by CHP. The phase transitions of the microsomal phospholipids influence the interaction of cytochrome P-450 with its reductase, altering the activation energy of type I substrates oxidation. The type II substrate oxidation is not affected by phase transitions in the full microsomal hydroxylating system.

Animals↗
PubMed1978

[Kinetics of N-dealkylation of amines with participation of microsomal cytochrome P-450].

Within the temperature range of 20-37 degrees C the kinetics of the demethylation reactions of a variety of amines with participation of hepatic microsomal cytochrome P-450, NADPH and O2 has been studied. Catalytical rate constants for all the substrates have been determined in generalized forms. Activation parameters deltaH* and deltaS* are also estimated. There is a linear relationship between deltaH* and deltaS*: deltaH*=20.7 kcal+366 degrees K deltaS*. Compensation relationship is characterized by a coefficient alpha=366 degrees/Taverage=1.21. The nature of the limiting step of the enzymatic amine demethylation involving cytochrome P-450 of hepatic microsomes is discussed.

Amines↗
PubMed1978

[Spectral parameters of the cytochrome c and hemoglobin interaction with methanol and aniline].

At 20 degrees C, in a phosphate buffer, pH 5,8--8,0, methanol and aniline interactions with hemoglobin and cytochrome c were studied using the difference spectrophotometry method. The difference absorption spectra are characterized by following values of lambdamax and lambdamin (nm): I--MeOH--hemoglobin (405 and 420), II-MeOH--cytochrome c (405--406 and 419--422), III--aniline--cytochrome c (421--410 and 401--396). The values of lambdamax and lambdamin for system III are shifted in the region of shorter wavelengths from 421 to 410 nm and from 401 to 396 nm, respectively within the pH range of 5,8--7,95. From difference spectra for systems I, II, III the dissociation constants of complexes obtained, Ks were calculated. Log Ks is linearly dependent on pH. System I is characterized by two values of Ks at all pH. The Ks values were calculated in general form from the dependences obtained. The nature of the complexes is discussed.

Aniline Compounds↗
PubMed1978

[Interaction of aliphatid alcohols with cytochrome P-450 from rat liver microsomes].

The interaction of alyphatic alcohols and cyclohexanol with cytochrome P-450 in microsomes has been investigated. All alchohols induced the modified 11 type spectral changes by mixing with microsomes. These changes are characterized by lambdamax = 412 and lambdamin = 380-382 nm in difference spectra. The dissociation constants of the alcohol cytochrome P-450 complexes are determined. On this dissociation constants influence pH and Triton X-100 presence. The interaction of the alcohols with cytochrome P-450 in phosphate buffer pH = 6,0 in the detergents absence is characterized by one dissociation constant for MeOH, EtOH, n-BuOH and cyclohexanol and by two dissociation constants for i-PrOH, i-BuOH and tert.-BuOH. The interaction of the alcohols with cytochrome P-450 in Tris-HCL-buffer (pH 7.5) in the Triton X-100 presence is characterized for all above alcohols by the dissociations constants, which are described by Taft equation with coefficient rho =-1.55. This fact confirms the interaction of alcohols HO-groups with heme iron of cytochrome P-450. The scheme of interaction of alcohols with cytochrome P-450 is discussed.

Animals↗
PubMed1978

[Kinetic and spectral parameters of the amines oxidative N-dealkylation with participation of the liver microsomal cytochrome P-450. Amines oxidation with organic hydroperoxides].

Kinetics of demethylation of a number of amines involving hepatic microsomal cytochrome P-450 and organic hydroperoxides (tret-butyl- and cumylhydroperoxide) have been investigated. Decomposition rate constants for the substrate-cytochrome P-450-ROOH complexes have been determined in a generalized form. Activation parameters, deltaH* and deltaS*, are calculated for decomposition of the complexes. There is a linear relation between deltaH* and deltaS*: deltaH*=18.7 kcal + 333 degrees K deltaS*. Compensation relationship is characterized by the value of alpha=333 degrees K/Taverage=1.11. The nature of the limiting step in the cytochrome P-450-NADPH-O2-system and the cytochrome P-450-ROOH-system is discussed.

Amines↗
PubMed1978

[Analysis of the interaction of glucocorticoid receptors obtained from rat liver with DNA in a aqueous dextran-polyethylene glycol biphasic system].

The binding of the receptor-glucocorticoid complexes from rat liver cells to DNA has been studied in an aqueous dextran-polyethylene glycol biphasic system. It has been shown that the capacity of DNA for binding hormone-receptor complexes is very large, and that no saturation is observed at physiological concentrations of the glucocorticoid receptor. The linear binding curve is indicative of low affinity of the DNA acceptor sites for the complexes. Cell cytosol contains substances which inhibit the binding of the receptor-dexamethasone complexes to homologous DNA.

Animals↗