[A comparative study of the nonhistone chromosomal proteins (pp23) from rat and bull kidneys]. / Sravnitel'noe izuchenie khromosomnykh negistonovykh belkov (pp23) iz pochek krysy i byka.

The bovine nonhistone protein pp23 differs from that of rat origin both immunologically (precipitation, specific interaction with monoclonal antibodies to rat pp23) and functionally (capability to stimulate expression of hetero-organic antigens on the membrane of intact hepatocytes cultured in suspension). These differences, however, are not recognized in the experiments on stimulation of DNA synthesis in Swiss 3T3 cell resting culture, when a more general cell function (proliferation) is dealt with, rather than a narrow specific one, such as synthesis of tumor-associated hetero-organic antigen of kidney origin.

[The isolation and characteristics of a protein from human and bovine kidneys close in molecular and biochemical properties to non-histone chromosomal antigen PP23]. / Vydelenie i kharakteristika belka iz pochek cheloveka i byka, blizkogo po molekuliarno-biokhimicheskim svoistvam NGB-antigenu PP23.

By ion-exchange gradient chromatography, autophosphorylated protein fractions were separated from nonhistone chromosomal protein (NHCP) samples isolated from bovine and human kidney and liver. Fractions, eluted from the phosphocellulose column with 0.4-0.5 M NaCl, could occur only in the case of kidney rather than liver NHCP-fractionation. The molecular weight of the major phosphorylated protein component, eluted in this zone, is about 20-26 kDa, which is close to the molecular weight of protein pp23, previously observed during similar fractionation of NHCP from rat kidney rather than from liver.

[The isolation of monoclonal antibodies to hepatoma-associated nonhistone chromosomal proteins from rat kidneys]. / Poluchenie monoklonal'nykh antitel k negistonovym belkam khromatina pochek krys, assotsiirovannym s gepatomami.

A monoclonal antibody (5D2) has been produced to a fraction of nonhistone proteins of kidney origin, associated with the rat hepatoma 27 and not obtained in normal liver. The 5D2 antibody was found to precipitate three proteins with molecular weights of 22-23 kDa. Indirect immunofluorescence studies on fixed hepatoma cell-preparations showed speckled nuclear staining patterns. In contrast, the 5D2 antibody was unreactive with intact hepatocytes.

[The fractionation of non-histone chromosomal proteins from rat kidney and liver and their DNA complexes by using the cation exchangers phosphocellulose and SE-Sephadex]. / Fraktsionirovanie khromosomnykh negistonovykh belkov pochek i pecheni krys i ikh kompleksov s DNK s ispol'zovaniem kationitov fosfotselliulozy i SE-sefadeksa.

Results of the gradient chromatography of chromosomal nonhistone proteins of the rat liver and kidney and their complexes with DNA on the phosphocellulose and SE-Sephadex are presented. The profiles of elution of the preparation of the homologous tissues with NaCl linear gradient of 0.1-1.0 M are equal. In the fractions of 0.4-0.5 M NaCl, protein components are discovered only in the samples of kidney.

[The expression of hetero-organic antigens of kidney origin on the surfaces of cultured rat hepatocytes under the influence of the narrow-band fractions of non-histone chromosomal proteins]. / Ekspressiia geteroorgannykh antigenov pochechnoi prirody na poverkhnosti kul'tiviruemykh gepatotsitov krys pod vliianiem uzkikh fraktsii khromosomal'nykh negistonovykh belkov.

Narrow fractions of nonhistone chromosomal proteins (NHCP) eluted with 0.4-0.5 M NaCl from the phosphocellulose column stimulate expression of hetero-organic antigens of kidney origin on the membrane of intact hepatocytes cultured in suspension. These fractions of NHCP were isolated from the intact rats kidney, from cells of hepatoma 27 and Zajdela hepatoma, and from the carcinogenic liver after a single diethylnirozamine injection. The membrane hetero-organic antigens were identified by means of indirect immunofluorescence using specific immune serum.

[The effect of preparations of non-histone chromosomal proteins on the expression of hetero-organic membrane antigens of kidney origin in a primary rat hepatocyte culture]. / Vliianie preparatov negistonovykh belkov khromatina na ékspressiiu membrannykh geteroorgannykh antigenov pochechnoi prirody v pervichnoi kul'ture gepatotsitov krys.

The expression of membrane hetero-organic antigens of kidney origin, associated with the rat hepatomas in primary culture of intact adult rat hepatocytes, was investigated by means of the indirect immunofluorescence method using a specific immune serum. These antigens were observed on the membrane of some hepatocytes after their contact with nonhistone chromosomal proteins (NHCP), which were obtained from the kidney of intact rats from cells of hepatoma 27 and Zajdela hepatoma, or from the carcinogenic liver after a single diethylnitrosamine injection. Negative results were obtained after the incubation of hepatocytes in the medium lacking some of NHCP, or in that with NHCP obtained from the liver of intact rats.

[The stimulation of DNA synthesis in resting Swiss 3T3 cells by non-histone chromosomal proteins]. / Stimuliatsiia sinteza DNK v pokoiashchikhsia kletkakh Swiss 3T3 negistonovymi khromosomal'nymi belkami.

Nonhistone chromosomal proteins (NHC), isolated from the kidney and liver of intact rats, the liver of rats treated with hepatocarcinogen DEN and the rat hepatoma, stimulate DNA synthesis in Swiss 3T3 cells in resting culture. The maximum stimulating effect was obtained in the presence of narrow NHC fractions eluted with 0.4-0.5 M NaCl from the phosphocellulose column and identified as hetero-organic NHC protein antigens of the kidney origin associated with hepatocellular tumors.

[Molecular weights of the hetero-organic NHCP antigens of kidney origin associated with rat hepatomas]. / Molekuliarnye massy assotsiirovannykh s gepatomami krys geteroorgannykh NGB-antigenov pochechnoi prirody.

The narrow NHCP protein fractions, possessing a proper phosphoprotein kinase activity, were isolated from kidney of intact rats, hepatoma tissue and liver cells of rats treated with hepatocarcinogen in the process of phosphocellulose gradient chromatography by elution of 0.4-0.5 M NaCl. The gel filtration on Sephadex G-75 and SDS-PAAG electrophoretic data demonstrate that all the NHCP protein fractions mentioned above include a general molecular component with the mass of 23 kD, and display identical antigenic properties. Thus, in accordance with the data obtained, the role of the hetero-organic NHCP protein antigen of kidney origin associated with hepatoma may be played by the general molecular component of NHCP protein fractions possessing properties of a specific chromosomal phosphoprotein kinase.

[Amino acid specificity of the phosphoprotein kinase activity of various fractions of non-histone chromatin proteins from the kidney, hepatoma and liver of rats after a single dose of diethylnitrosamine]. / Aminokislotnaia spetsifichnost' sobstvennoi fosfoproteinkinaznoi aktivnosti nekotorykh fraktsii negistonovykh belkov khromatina pochek, gepatomy i pecheni krys pri odnokratnom deistvii diétilnitrozamina.

Amino acid specificity of phosphorylation of some nuclear non-histone proteins was studied in the fractions, eluted by 0.4-0.5 M NaCl from phosphocellulose columns, using the reaction of 33P transfer, catalized by native phosphoprotein kinase, from gamma-33P-ATP into the proteins. A number of amino acid residues, capable to reaction, were phosphorylated in the 0.4-0.5 M NaCl fraction of non-histone proteins from kidney of intact rats, from cells of hepatoma 27 and from liver cells after a single intraperitoneal administration of diethylnitrosamine. At the same time, methionine residue was markedly phosphorylated in the fraction of 0.4-0.5 M NaCl of intact rat kidney, whereas this effect was not found in the preparations of hepatoma 27 and in cells of "carcinogenous" liver tissue. As distinct from kidney non-histone proteins, proteins from the 0.4-0.5 M NaCl fraction of the latter preparations tended to exhibit tyrosine phosphokinase activity. It is well known that many of oncoproteins exhibited mainly the tyrosine-specific phosphoprotein kinase activity. The data obtained suggest that "tyrosine" shift in the preparations of non-histone proteins from tumoral cells or from rat liver cells, altered under the effect of hepatocarcinogenesis, may be an indication of functional activity of oncogenes.

[Radiometric detection of nonvisualizable immunoprecipitates formed by hepatoma-associated hetero-organic renal antigens of 33P-phosphorylated fractions of non-histone chromatin proteins]. / Radiometricheskoe vyiavlenie nevizualiziruemykh immunopretsipitatov, obrazuemykh assotsiirovannymi s gepatomami geteroorgannymi antigenami pochechnoi prirody 33P-fosforilirovannykh fraktsii negistonovykh belkov khromatina.

A study was made of the antigenic properties of nuclear nonhistone proteins (NHP) from rat tissue eluted with 0.4-0.5 M NaCl gradient on phosphocellulose at very low concentrations. 33P-labeled proteins were obtained by phosphorylation of NHP fractions in the presence of [gamma-33P] ATP. Some non-visual precipitates which appeared during the reaction of specific inhibition of precipitation in agarose gel of 33P-labeled NHP-fractions from rat solid hepatoma 27 cells or rat liver after single hepatocarcinogen diethylnitrosamine injection were detected by means of a calculable radiometric method, proposed by the authors. These precipitates appear only if the rabbit immune sera against NHP-DNA complexes of intact rat kidneys exhausted with liver NHP-antigens was used, never appearing in the case of kidney NHP-antigen exhaustion. Therefore the NHP-fractions prepared from "carcinogenic" liver and rat solid hepatoma cells which possess proper phosphoprotein kinase activity and form specific immune precipitates must be identified as hetero-organic antigens of kidney origin.

[Hetero-organic antigens of renal nature in the composition of the nuclear nonhistone proteins in the regenerating rat liver]. / Geteroorgannye antigeny pochechnoi prirody v sostave iadernykh negistonovykh belkov regeneriruiushchei pecheni krys.

In the nuclear non-histone proteins of the rat liver, on the 4th day after a partial hepatoectomy or hepatocarcinogen injection as well as in the hepatocellular tumour cells, some heteroorganic antigens of kidney nature (HAkid) are found and characterized immunochemically. These HAkid can be eluted at 0.4-0.5 M NaCl during gradient chromatography on phosphocellulose. They possess some proper phosphoproteinkinase activity. The appearance of HAkid in rat liver may be considered, on the one hand, as a manifestation of some malignant factor (carcinogen action, tumour cells) and, on the other hand, they are obviously connected with proliferative activity of the regenerative rat liver cells (hepatectomy effect). It is very likely that both the sides of this phenomenological effect of HAkid in rat liver are the consequence of a specific expression along the cell oncogens.

[Detection of the common features in the antigenic structure and the spectra of the phosphoprotein kinase activity proper of the water-soluble fractions of DNA-free nuclear nonhistone proteins from hepatomas and the liver in rats following a single administration of hepatocarcinogens]. / Vyiavlenie obshchikh chert v antigennoi strukture i spektrakh sobstvennoi fosfoproteinkinaznoi aktivnosti vodorastvorimykh fraktsii svobodnykh ot DNK iadernykh negistonovykh belkov gepatom i pecheni krys posle odnokratnogo vvedeniia gepatokantserogenov.

By means of the reaction of specific inhibition of precipitation in agarose gel and using the rabbit immune sera against nuclear nonhistone proteins (NHP) in the NHP-DNA complexes isolated from the rat kidney it is shown that the single intraperitoneal injection of hepatocarcinogens, 4-dimethylaminoazobenzene (DAB) or N-nitrosodiethylamine (DEN) induces the appearance of hetero-organic antigens of the kidney nature in the NHP pattern of the liver. These antigens identical, as it proved to be, to the same NHP-antigens from the cells of transplantable rat hepatoma 27 and ascitic Zajdela hepatoma could be found in the NHP pattern of the rat liver during 1 to 12 and 1 to 64 days after the DAB and DEN injection, respectively. In all cases when the hetero-organic NHP-antigens were found the profiles of proper phosphoprotein kinase activity of the fractions of NHP eluted by 0.4-0.5 M NaCl using gradient chromatography on phosphocellulose contained peaks that were not characteristic to the liver NHP but were typical of NHP from the intact rat kidney. In the SDS-PAAG electrophoresis the mentioned fractions form one line with the molecular weight 15000-20000.

[Isolation of nonhistone proteins from animal cellular chromatin]. / O vydelenii negistonovykh belkov iz khromatina kletok zhivotnykh.

Difficulties of various methods of isolation of DNA-free non-histone proteins (NHP) from different animal tissues and tumor cells are discussed. As a result, a simple and effective way of obtaining clean preparations of NHP possessing phosphoproteinkinase and immunological activity has been described.

[Comparative study of the immunochemical properties of the 5th lactate dehydrogenase isoenzyme from rat liver and skeletal muscle. II. Isolation and characteristics of LDH-5 preparations from rat liver. General analysis of the results]. / Sravnitel'noe izuchenie immunokhimicheskikh svoistv piatogo izozima laktatdegidrogenazy iz skeletnykh myshts i pecheni krys. II. Poluchenie i kharakteristika preparatov LDG-5 iz pecheni krysys. Obshchii analiz rezul'tatov.

The method of isolation of chromatographically homogenic LDG-5 preparation from the rat liver was elaborated. The preparations obtained in the present study and LDG-5 preparations from skeletal muscles have a common component which is testified by results of the precipitation test in agar. In the rat liver there is an enzymatically non-active antigenic component, which, according to some of its physico-chemical parameters, is very similar with chemically modified forms of LDG-5 dimers.

[Conformation of trypsin molecules in aqueous solutions containing 2-chloroethanol]. / Konformatsiia molekul tripsina v vodnykh rastvorakh, soderzhashchikh 2-khlorétanol.

Changes in the macromolecular parameters of trypsin in the presence of 2-chloroethanol in aqueous solutions have been studied by means of optical and hydrodynamic methods. At temperature--dependent volume fraction of 2-chloroethanol in solution upsilon t < 0.30 the globular structure of trypsin is destroyed but the regularity of polypeptide chains within the limits of secondary structure is maintained. At 0.30 < upsilon < 0.80 the solvation envelope of macromolecules is kept constant mainly owing to hydration, but the solubilization takes place only at upsilon < 0.30. At upsilon < 0.80 spiralization sharply increases and reaches in pure 2-chloroethanol its maximum value (50%). The intrinsic viscosity moreover reaches only half the whole value [eta]coil--[eta]glob.

[Comparative study of the immunochemical properties of the 5th lactate dehydrogenase isoenzyme from rat skeletal muscles and liver. I. The isolation and characteristics of LDH-5 preparations from skeletal muscles]. / Sravnitel'noe izuchenie immunokhimicheskikh svoistv piatogo izozima laktatdegidrogenazy iz skeletnykh myshts i pecheni krys. I. Poluchenie i kharakteristika preparatov LDG-5 iz skeletnykh myshts.

Individual LDH-5 preparations from rat skeletal muscles have been obtained by differential fractionation with (NH4)2SO4 and precisional ion exchange chromatography. According to the data of ultracentrifugation, electrophoresis in acrilamide gel, enzyme-electrophoresis and agar precipitation reaction with antimuscle serum, these preparations have no admixtures of other isoenzymes of foreign proteins. Examination of antigenic properties of these LDH-5 preparations may suggest the availability of common antigenic determinates in the liver extract and in the LDH-5 preparation.