PubMed Health⌕ Search

Biomedical subjects

O V Sviridov

Publications and source records attributed to O V Sviridov.

At least 19 recordsLinked to original sources
PubMed1994

[A new property of known proteins: specific binding of thyroid hormones by human plasma apolipoproteins].

The thyroid hormones--thyroxine (T4) and triiodothyronine (T3)--are capable of associating with human plasma high (HDL), low (LDL) and very low density lipoprotein particles (VLDL). Apolipoproteins (apo) act as the hormone-binding components of the lipoprotein particles. The thyroid hormone binding to apolipoproteins is a time-dependent, reversible, saturable and sensitive to specific inhibitors interaction with the structurally isolated site in the protein which is complementary to the ligand. The number of such sites per macromolecule varies from one (apoA-I) to three (apoB-100). Low affinities (Ka approximately 10(5)-10(6) M-1) for T4 are characteristic of apoA-II, apoA-IV, apoE and apoB-100, while apoA-I and its lipid complex, apoA-I-HDL display sufficiently high affinities for the hormone (Ka approximately 10(7)-10(8) M-1). The active site of apoA-I-HDL contains a chemical group interacting with the ionized phenolic ring hydroxy-group of T4, includes cavities capable of accommodating iodine atoms of the ligand molecule, has a hydrophobic nature and low stereospecificity and is located close to the surface of the protein globule in the N-terminal part of polypeptide chain conformationally associated with the polar surface monolayer of the lipid matrix. Three hormone-binding sites in apoB-100 are located in different regions of the polypeptide chain which are distant from each other and lie outside the sites of heparin and cellular LDL receptor binding. ApoB-100 and apoE stimulate T4 entry into fibroblasts, while apoA-I inhibits the interaction of T4 and T3 with human placental plasma membranes.

Apolipoproteins↗
PubMed1994

[Interaction of thyroid hormones with immunoglobulins isolated from human blood serum. I. Parameters of complex formation and the nature of the binding reaction].

The kinetic and equilibrium characteristics of the interaction of thyroxine (T4) with immunoglobulins (Ig) A, G and M as well as with Bence-Jones proteins purified from human blood serum have been investigated. The formation of complexes between T4 and human immunoglobulins has been found to be time-dependent, reversible, saturable and sensitive to specific inhibitors. The L-chain (ae or lambda) is a component of the immunoglobulin molecular structure which appears to be essential and sufficient for the T4 binding. The covalent attachment of the H-chain can increase dramatically the affinity for the thyroid hormone (the mu-chain in IgM) or alter the sensitivity of the binding region to chemical agents and pH (the mu-chain in IgM, the gamma-chain in IgG). The experimental data suggest that the T4-binding IgM does not belong to a pathological type of proteins- anti-T4 autoantibodies-because: (i) the dependence of the T4 binding reaction on the physico-chemical conditions of the environment is typical of normal transport proteins; (ii) the prevalence of the T4-binding IgM in random individual serum samples from healthy subjects is 100%; (iii) the IgM-T4 complex differs structurally from the antigen-antibody complex, being unable to interact with the first complement component. The specific T4-binding properties of normal human serum immunoglobulins could remain so far unrecognized due to the inability of the conventional analytical methods to detect the weak manifestations of the T4-binding activity of these proteins in physiological fluids containing the endogenous inhibitor (Cl-), and/or the exogenous inhibitor (8-anilino-1-naphthalene sulphonic acid).

Autoantibodies↗
PubMed1994

[Interaction of thyroid hormones with immunoglobulins isolated from human blood serum. II. Structural elements and localization of thyroxine-binding segment in the immunoglobulin M molecule].

The structural characteristics and topography of the thyroxine T4-binding site on the human immunoglobulin M (IgM) molecule have been studied using affinity binding with T4 structural analogs blocking the formation of the T4-IgM complex with proteins displaying an affinity for individual structural components of IgM and proteolytic fragmentation followed by determination of the T4-binding activity of the isolated fragments. It has been found that IgM has a poor selectivity towards the binding of T4 structural analogs which is characteristic of transport proteins but not of autoantibodies. The T4-binding region of IgM lies outside the variable portion of the Fab-fragment and is apparently constituted by the Cmu 1-domain of the heavy chain and the constant part of the light chain linked via a disulphide bridge. The T4-binding site located within this region possesses no stereospecificity and contains structural elements complementary to the iodine atoms of the outer ring and the aliphatic chain of the iodothyronine molecule.

Binding Sites↗
PubMed1993

[Structural changes of lipid and thyroxine-binding protein zones of lipoprotein particles, containing human apolipoprotein A-1].

The relationship between temperature-induced structural changes in the lipid and protein thyroxine-binding zones of lipoprotein particles containing human apolipoprotein A-1 (apoA-1) as a sole protein component was studied using ESR with probes of two types--spin-labeled thyroxine and 5- or 16-doxylstearic acids. A structural rearrangement in the protein active site at 23-24 degrees was found. In the same temperature range, a dramatic change in the monolayer phospholipid ordered structure was observed. Based on these data, the localization of the apoA-1 thyroxine-binding site in the lipid phase as well as the possibility of regulation of the hormone binding process by changes in the protein-lipid interaction in lipoprotein particles are discussed.

Apolipoprotein A-I↗
PubMed1993

[Simulating and inhibiting effect of individual blood serum transport proteins on thyroid hormone binding with human placental cell membrane].

Binding processes in in vitro systems modelling specific interactions between transport and receptor proteins, thyroid hormones of human placental tissue and the washing blood, have been studied. These systems included syncytiotrophoblast villous membranes, thyroxine (T4), triiodothyronine (T3) and iodothyronine-binding proteins: transthyretin, albumin, immunoglobulins (Ig) M and G, apolipoprotein A-I, and the T4-binding globulin purified from human retroplacental serum. All of the transport proteins at concentrations close to the Ka values of their complexes with thyroid hormones produced inhibitory effects on the binding of [125I]T3 or [125I]T4 to the thyroid hormone membrane receptor. The dependence of [125I]T4 membrane binding on IgM concentration in the system was characteristic of all proteins studied. In the case of T3 such dependence was unique for IgM, and included the phase of the IgM stimulatory effect (10(-11)-10(-9) M) and the phase of inhibition (10(-8)-10(-7) M). In the presence of 30 pM IgM, the concentration of the membrane T3-binding sites increased by 75% with a 2.2-fold decrease of the association constant (Ka). It was shown that IgM interacts specifically with two classes of binding sites on the plasma membranes with Ka(1) = 5.0 x 10(9) M-1, Bmax(1) = 34 fmol/mg of membrane protein and Ka(2) = 2.7 x 10(7) M-1, Bmax(2) = 2.0 pmol/mg of membrane protein. It is suggested that the stimulatory effect of IgM is caused by increases in the number of T3-binding sites on the placental villous membranes as a result of complex formation between IgM and its membrane receptor which displays an enhanced T3-binding activity.

Binding Sites↗
PubMed1992

[Thyroid hormone conjugates with rhodamine B as fluorescent ligands of human plasma transport proteins].

Conjugates of thyroxine (T4) and triiodothyronine (T3) with rhodamine B in which the hormone and the fluorescent dye are linked via a thiourea bond have been synthesized. These conjugates possess an ability to inhibit in a competitive manner the binding of [125I]T4 to three protein preparations: T4-binding globulin (TBG), apolipoprotein A-I (ApoA-I), and high density lipoprotein particles (ApoA-I-HDL) isolated from human serum by T4-Sepharose 4B chromatography and further purified. The following values of association constants have been estimated: for the T4 derivative-3 x 10(7) M-1 (TBG), 4.1 x 10(5) M-1 (ApoA-I), and 4.2 x 10(5) M-1 (ApoA-I-HDL); for the T3 derivative-1.6 x 10(7) M-1 (TBG), 5.3 x 10(5) M-1 (ApoA-I), and 5.4 x 10(5) M-1 (ApoA-I-HDL). The binding of rhodamine B-labeled thyroid hormones to TBG or ApoA-I do not alter significantly the parameters of rhodamine B chromophore absorption and fluorescence. The interaction of the conjugates with ApoI-HDL leads to a significant enhancement of the absorption intensity and a 3 nm blue shift in the absorption maximum as well as to a 1.5-fold increase in the fluorescence band amplitude at 586 nm. Biological and fluorescent properties of T4 and T3 derivatives suggest that these compounds may be a useful tool in fluorescence studies of plasma binding protein-driven transport of thyroid hormones in model biological systems.

Binding, Competitive↗
PubMed1991

[Isolation, identification and properties of a new thyroxine-binding protein--apolipoprotein A-1 from human blood serum].

A protein having a molecular mass of about 25 kWa was isolated by thyroxin (T4)-Sepharose affinity chromatography from human blood serum; its properties were found to be distinct from those of known T4-binding proteins. Using immunodiffusion, radioimmunoassay, lipid analysis, differential precipitation and electrophoresis, it was shown that the isolated protein is a component of high density lipoprotein (HDL) particles and represents an apolipoprotein A-1 (apoA-1). Using cholate-Sepharose chromatography apoA-1 was separated from the lipid moiety and contaminant proteins, and apoA-1 was effectively isolated directly from the blood serum. Apo-A-1-HDL and apoA-1 obtained by affinity chromatography as well as the HDL3 fraction isolated by a standard ultracentrifugation technique, all displayed a T4-binding activity, the affinities for the hormones being of the same order of magnitude. The T4 interaction with these preparations induced difference UV-absorption signals, altered the characteristics of apoA-1 intrinsic fluorescence without affecting the circular dichroism of the protein-hormone system. The binding of spin-labelled T4 to apo-1, apoA-1-HDL or HDI3 caused substantial changes in the shape of the ESR spectrum and an increase in the apparent rotational correlation time. The mobility of the radical fragment of spin-labelled T4 depended on the composition and properties of the protein preparation. The electron spectroscopy data suggest that the T4-HDL interaction occurs via specific mechanisms and that the molecular structures of the complexes formed thereby are not characteristic of other known T4-binding proteins.

Apolipoprotein A-I↗
PubMed1990

[Affinity chromatography of thyroxine-binding proteins from human serum].

The affinity matrix prepared by the attachment of L-thyroxine (T4) to epichlorohydrine-activated Sepharose 4B biospecifically absorbs the T4-binding globulin (TBG), 25K and 80/27K proteins, immunoglobulin G (IgG) and albumin (HSA) from human normal and retroplacental sera. The absorbed protein patterns were shown to depend on the immobilized T4 concentration, pH, temperature and incubation time. The potent eluents desorbing 85-100% of the protein are 1 mM NaOH, 3 M NH4SCN, 10(-5) M T4 or 3 mM 8-anilinonaphthalene-1-sulfonic acid (ANS) for TBG; NaOH, NH4SCN, 3 mM MgCl2 or 12mM sodium cholate for 25K protein and HSA; NaOH, NH4SCN or MgCl2 for the 80/27K and 25K proteins and IgG. Moreover, T4 desorbs small amounts (6-8%) of the 80/27K and 25K proteins, while sodium cholate elutes about 6% of TBG. The eluted from T4-Sepharose 4B and further purified TBG, 25K and 80/27K proteins display different [125I]T4-binding activities within the pH range from 2 to 9 and differ by their resistance to thermal inactivation at 50-80 degrees C. Double radial immunodiffusion analysis with the use of antisera to TBG, 25K, 80/27K, HSA and IgG demonstrated that the proteins share no common antigenic determinants. It was concluded that the novel 25K and 80/27K proteins represent endogenous components of the human blood thyroid hormone-binding protein system rather than fragments or aggregates of the known T4-binding proteins.

Chromatography, Affinity↗
PubMed1990

[Specific binding of iodothyronines with apolipoprotein A-1 in high density lipoprotein particle isolated from human serum by affinity chromatography on thyroxine-sepharose].

The kinetic and equilibrium characteristics of interaction of thyroxine (T4) and its structural analogs with a high density lipoprotein (HDL) fraction isolated from human serum by T4-Sepharose affinity chromatography and containing apolipoprotein A-I (apo A-I) as a sole protein component, were studied. The binding of [125I]T4 to apo A-I-HDL reached a maximum after 40 min and did not change during the next 80 min of incubation at 0 degrees--22 degrees C. Dissociation of [125I]T4 induced by the addition of excess unlabeled T4 to the complex solution proceeded more intensely on a time scale at 0--2 degrees C than at 22 degrees C. Incubation of apo A-I-HDL with increasing concentrations of T4 showed that the binding is saturable. The data analysis using different computer programs revealed the presence in apo A-I-HDL of a single class of binding sites with K alpha = (4.0 +/- 2.1).10(-7) M- and Bmax = 1.7 +/- 0.8 nmol T4/mg of protein. Naturally occurring iodothyronines, their analogs and D-isomers of thyroid hormones competed with [125I]T4 for the binding sites on apo A-I-HDL with the following inhibitory potencies: L-T4 = D-T4 greater than or equal to 3,3',5-triiodo-L-thyronine = 3,3',5-triiodo-D-thyronine greater than 3,5-diiodo-L-thyronine = 3,3',5- triiodothyroacetic acid greater than 3,3',5-triiodothyropropionic acid greater than or equal to 3,5-diiodo-L-thyrosine.(ABSTRACT TRUNCATED AT 250 WORDS)

Apolipoprotein A-I↗
PubMed1989

[New aspects of isolation and purification of thyroxine-binding globulin from human biological fluids].

Based on the new data concerning the multicomponent system of thyroxine-binding proteins in human plasma, some methodological aspects of isolation and purification of thyroxine-binding globulin (TBG) were examined, and a simple two-step procedure for TBG purification was developed. Normal human blood serum, retroplacental serum and amniotic fluid were used as TBG sources. The procedure includes affinity chromatography and adsorption chromatography on a hydroxyapatite column. A biospecific adsorbent was synthesized by stepwise binding of epichlorohydrin and thyroxine to Sepharose. The yield of pure TBG varied from 60 to 80%, depending on the TBG source used. The properties of TBG preparations from retroplacental serum and amniotic fluid were identical; both preparations contained a pregnancy-associated molecular variant, TBG-1. Two novel serum thyroxine-binding proteins were detected, isolated and partly characterized.

Amino Acids↗
PubMed1988

[Effect of structural changes in thyroxine-binding globulin on its biological activity and immunochemical properties].

Using spectroscopic, electrophoretic and microcalorimetric techniques, the changes in the spatial structure of human thyroxine-binding globulin (TBG) induced by exposure of protein solutions to high temperatures (45-90 degrees C) and low pH (2.5-6.0) were studied. Simultaneously the biological activity and immunoreactivity of TBG samples were measured. The structural changes were manifested at 52 degrees C or at pH 4.0 and were then aggravated with a rise in temperature or a decrease of pH. The circular dichroism spectra showed that the molecular ellipticity had a maximum decrease (by 10%) at 218-222 nm. In fluorescence spectra excitable at 280 nm the band half-width increased by 4-6 nm; their intensity decreased by 30-40%, whereas the position of the maxima did not change significantly. After addition of an equimolar amount of thyroxine to inactivated TBG the protein fluorescence was quenched by 25-40%. The electrophoregrams of treated preparations contained additional protein bands possessing no biological activity, whose mobility was less than that of native TBG. Microcalorimetric assays of native TBG revealed a thermoabsorption peak with a maximum at 62.5 degrees C and a half-width of 7.1 degrees C. The thermodynamic parameters of melting of TBG spatial structure were consistent with a model of a two-domain structure of the molecule. The biological activity and immunoreactivity of TBG showed a coordinated decrease with a rise in the degree of protein denaturation, However, the formation of TBG complex with antibodies did not screen the thyroxine-binding center of TBG and did not alter its affinity. Possible mechanisms of structural transition of TBG and its effect on the biological properties of TBG are discussed.

Binding Sites, Antibody↗
PubMed1986

[Radioimmunologic system for the quantitative determination of thyroxine in human blood serum].

A methodological approach to development of a radioimmunological system for quantitative assay of thyroxine (T4) in human blood serum is described. It implies investigation of every component of the system and matching of the components by the physico-chemical properties and concentrations. An antiserum to the T4 conjugate was prepared with bovine serum albumin. The hapten "fitting density" was 30 mol per 1 mol of protein. The antiserum had a titer of 5000, was highly specific and showed affinity to T4, Ka (7.6 +/- 1.0) X 10(8) M-1. The characteristics of the antiserum did not depend on concentration of the hydrogen ions in solution at pH 6.0-9.0 and the ionic strength of the buffer solutions within M 0.05-1.0. 8-Aniline-1-naphthalene sulfonic acid (ANA) in concentrations of 0.05-0.1 per cent completely inhibited binding of T4 to endogenic proteins of the blood serum and had no effect on the antiserum interaction with T4. This provided performance of the radioimmunological assays at 0.1 per cent concentration of ANA. [125I]T4 with the specific activity of 40 TBk/g was used in the study. The quality of [125I] T4 was estimated by its immune reactivity. The standard solutions of T4 were prepared with donor blood serum as the basis. It was shown that quantitative assays of T4 in blood serum within clinically significant ranges of the hormone concentration are possible at 1:5000 final dilution of the antiserum, the [125I] T4 activity of 20 000-30 000 imp/min per a specimen and 5 per cent content of the blood serum in the analytical specimen. A simple and reliable technique for T4 assay in human blood serum is developed.

Animals↗
PubMed1985

[Comparative characteristics of molecular forms of human thyroxine-binding globulin].

Two molecular variants, of thyroxin-binding globulin (TBG), TBG-1 and TBG-2, were obtained from human retroplacental blood by fractionation of pure TBG on concanavalin A-Sepharose. It was found that both variants are immunologically identical, have similar molecular weights, amino acid composition and spectral properties, and possess the same affinity for thyroid hormones. However, TBG-1 and TBG-2 differed in charge upon isoelectrofocusing and had different monosaccharide composition. The existence of two molecular variants of TBG in pregnancy is probably due to the peculiarities of the polypeptide chain glycosylation during TBG biosynthesis.

Amino Acid Sequence↗
PubMed1983

[Purification and physico-chemical properties of the sex steroid-binding globulin of human blood].

A new technique for purification of the human sex hormone-binding globulin (SHBG) is described. This technique includes affinity chromatography of blood serum on cortisol-Sepharose, (NH4)2SO4 fractionation, gel filtration on a Bio-Gel P-300 column and chromatography on a concanavalin A-Sepharose 4B column. From 21 of retroplacental serum 10 mg of pure SHBG (25% yield) has been obtained. Upon gel filtration SHBG behaved as a biopolymer with Mr of 120,000. The molecular weight of SHBG as determined by electrophoresis was shown to be equal to 50,000. SHBG has a sedimentation constant of s20, w of 4.7S, pI of 5.75, extinction coefficient A1%(280,1cm) = 10,5 and association constants of 4.5 X 10(8) and 3.5 X 10(6) M-1 for 5 alpha-dihydrotestosterone and cortisol, respectively. The amino acid and carbohydrate contents of SHBG were determined.

Chromatography, Affinity↗
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↗