[Characteristics of carbohydrate chains of two forms of salmon gonadotropic hormone binding and not binding concanavalin A]. / Kharakteristika uglevodnykh tsepei sviazyvaiushcheisia i ne sviazyvaiushcheisia s konkanavalinom A form gonadotropnogo gormona gorbushi.

A comparative analysis of carbohydrate chains of two forms of hunchback salmon gonadotropin that bind or not with ConA-Sepharose was carried out. It was found that unbound, "protein", ConA(-)-form contains three times less carbohydrates and has slightly different amino acid composition as compared to the bound, normally glycosylated ConA(+)-form. HPLC fractionation showed the oligosaccharides released from both hormone forms to be mainly sialylated. The major oligosaccharides identified in ConA(+)-form are biantennary, fucosylated (approximately 20%) or nonfucosylated; minor bissected chains are also present. In ConA(-)-form the same oligosaccharides and also oligosaccharides with a higher degree of branching (tri- and tetraantennary) were detected. These two hormone forms isolated from pituitary glands of male or female fish differed neither in the type of oligosaccharides nor in their ratio. Essential structural differences in carbohydrate chains of hunchback salmon and sturgeon gonadotropins were distinctly demonstrated.

Carbohydrates in mammalian tryptophanyl-tRNA synthetase.

Homogeneous preparations of bovine tryptophanyl-tRNA synthetase (EC 6.1.1.2) contain monosaccharides (mannose, fucose, galactose, N-acetylglucosamine) as revealed by liquid chromatography. Their content comprises 2.5-3.0% (w/w) of the enzyme composed of two subunits (60 kDa x 2). The same set of sugars was detected in elastase and CNBr-generated fragments (with molecular masses of approx. 40 kDa and 30 kDa, respectively). It is concluded that bovine tryptophanyl-tRNA synthetase, in addition to being a metallo- and phosphoprotein, is also a glycoprotein.

[The structure of carbohydrate chains of human IgG4-paraproteins differing in the ability to bind cell receptors]. / Struktura uglevodnykh tsepei IgG4-paraproteinov cheloveka, razlichaiushchikhsiia po sposobnosti sviazyvat'sia s kletochnymi retseptorami.

Comparative oligosaccharide analysis by HPLC revealed structural differences in the carbohydrate chains of human IgG4 paraproteins, varying in ability to induce the rhesus monkey's passive skin anaphylaxis. An atypical IgG4 paraprotein, which is inactive in this reaction and also does not bind the IgG4-subclass specific monoclonal antibody IH2, has a much higher proportion of the carbohydrate chains lacking terminal galactose residues than two typical IgG4 paraproteins. This structural feature may be one of the reasons for the atypical IgG4 not to bind by the mast cell Fc gamma receptor.

[A simple method of isolating monoclonal immunoglobulin M, possessing rheumatoid activity]. / Prostoi metod vydeleniia monoklonal'nogo immunobloulina M, obladaiushchego revmatoidnoi aktivnost'iu.

A simple procedure for obtaining highly purified preparations of native monoclonal (Waldenström's disease) immunoglobulin M possessing a rheumatoid activity (IgM-RF) has been developed. The method is based on the use of affinity chromatography with a new readily available adsorbent (immunoglobulin G-porous glass) and 3 M LiCl in Tris-buffer pH 8.3-8.4 able to induce the dissociation of the IgM-RF-IgG complex. The IgM-RF preparation thus obtained was characterized in terms of amino acid composition (relative to conventional monoclonal IgM), carbohydrate composition and structure of oligosaccharide moieties of a complex type. It was shown that some dissociation conditions for the IgM-RF-IgG complex routinely used to isolate IgM-RF provoke irreversible denaturation of IgM-RF when applied to a preliminarily purified complex.

[Structure of carbohydrate chains of dimeric molecules and individual subunits of gonadotropin from the Russian sturgeon]. / Struktura uglevodnykh tsepei dimernoi molekuly i otdel'nykh sub''edinits gonadotropina russkogo osetra.

Carbohydrate chains of gonadotropin from the Russian sturgeon hypophysis, as well as of alpha- and beta-subunits of the hormone, were split off and fractionated by gel-chromatography and HPLC. More than ten oligosaccharides released from the male and female hormones gave almost identical patterns, whereas differences between alpha- and beta-subunits were more noticeable. Basing on the chromatographic properties and monosaccharide compositions of the oligosaccharides isolated and the known structures of N-linked carbohydrates of mammalian hormones, the common carbohydrate chain of sturgeon gonadotropin is as follows: [formula: see text] Some oligomannosidic and/or hybrid chains and small oligosaccharides of the pentasaccharide core type were also found. Carbohydrate chains of fish gonadotropin have fewer sialic acid residues and significantly fewer (if any) sulphate groups than the mammalian hormones.

[The structure of carbohydrate chains of hemagglutinin and neuraminidase of influenza virus B/Leningrad/179/86]. / Struktura uglevodnykh tsepei gemaggliutinina i neiraminidazy virusa grippa B/Leningrad/179/86.

The main surface glycoprotein, hemagglutinin (HA), was obtained by treatment of influenza virus B/Leningrad/179/86 with bromelain. Amino acid and monosaccharide compositions of HA and neuraminidase (NA, earlier isolated from the same virus) were determined, thus showing HA and NA to contain 8-10 and 2 carbohydrate chains, respectively. The carbohydrate fragments were cleaved off by the alkaline LiBH4 treatment, the oligosaccharides released were reduced with NaB3H4 and fractionated by two-step HPLC on Ultrasphere-C18 and Zorbax-NH2 columns. Some higher mannose and complex oligosaccharides were identified in both cases by comparison with nonlabelled oligosaccharides of the known structure. The data obtained show that surface glycoproteins of influenza virus A and B are rather similar with regard to structure and heterogeneity of their carbohydrate chains.

[The structure of oligosaccharide fragments of glycoproteins from influenza virus A/Krasnodar/101/59/(H2N2)--heavy and light chains of hemagglutinin and neuraminidase]. / Struktura oligosakharidnykh fragmentov glikoproteinov virusa grippa A/Krasnodar/101/59(H2N2)--tiazheloi i legkoi tsepei gemaggliutinina i neiraminidazy.

The structure and heterogeneity of carbohydrate chains of hemagglutinin (HA) and neuraminidase (NA), the surface glycoproteins of influenza virus A/Krasnodar/101/59 (H2N2), were investigated. Hemagglutinin was reduced with beta-mercaptoethanol and its heavy (HA1) and light (HA2) chains were separated by gel chromatography. Amino acid and sugar composition of HA1, HA2 and NA was elucidated. The carbohydrate chains of the glycoproteins were cleaved off by the alkaline LiBH4 treatment and oligosaccharides were reduced with NaB[3H]4. They were fractionated by subsequent two-step HPLC on Ultrasphere-C8 and Zorbax-NH2 columns with simultaneous identification using nonlabelled oligosaccharides of known structures. Some of the major oligosaccharides isolated from HA1, HA2 and NA were thus identified as high mannose chains, containing 5-9 mannose residues, and complex chains, first of all biantennary chains having or not having bisecting N-acetylglucosamine and/or fucose residues. The approach which has been developed enables one to study the structure and heterogeneity of carbohydrate chains starting from one nmole of a desialylated N-glycoprotein.

[Effect of carbohydrate components of ristomycin A on platelet aggregation in human plasma]. / Vliianie uglevodnogo sostava ristomitsina A na agregatsiiu trombotsitov plazmy krovi cheloveka.

In the process of the investigation, conditions for specific removal of arabinose in tetrasaccharide of ristomycin A, a glycopeptide antibiotic as well as conditions for simultaneous removal of arabinose and mannose-2 bound to actinoidinic acid were determined. The role of arabinose in manifestation of the ristomycin A ability to induce platelet aggregation was shown to be important. Mannose-2 also had the same ability while its level was somewhat lower.

[The structure of complex carbohydrate chains of hemagglutinin from influenza viruses A/Kiev/59/79 (H1N1), A/Chile/1/83/25(H1N1) and X/79(H3N2)]. / Struktura uglevodnykh tsepei kompleksnogo tipa v gemaggliutinine virusov grippa A/Kiev/59/79(H1N1), A/Chili/1/83/25(H1N1) i X/79(H3N2).

An earlier developed method of identification of oligosaccharides by HPLC was used for studying the carbohydrate chains of three hemagglutinins from various influenza virus strains. The structures of main oligosaccharides of the complex type were elucidated on the basis of their chromatographic characteristics and monosaccharide composition. Oligosaccharide patterns varied in the above hemagglutinin samples but in all cases the major complex chains were fucosylated and nonfucosylated biatennary chains; bisected and triantennary chains were also found.

The carbohydrate chains of influenza virus hemagglutinin.

The major surface antigen of influenza virus A/Leningrad/385/80 (H3N2), H3 hemagglutinin, as well as its heavy and light subunits were obtained by bromelain treatment, followed by gel chromatography. Carbohydrate chains were split off from both subunits by lithium borohydride-lithium hydroxide in aqueous 2-methyl-2-propanol, and individual oligosaccharides isolated. The main oligosaccharides, whose structure was determined by 1H-n.m.r. spectroscopy and chemical methods, are of the ordinary oligomannoside and complex types. It was found that, in spite of the great difference in number of glycosylation sites in heavy and light subunits, the amount and even relative abundance of variants of carbohydrate chains in both subunits are very similar.

[Comparative study of carbohydrate chains of H1 hemagglutinins of influenza viruses A/Kiev/59/79 and X/Leningrad/54/1 using oligosaccharide maps]. / Sravnitel'noe izuchenie uglevodnykh tsepei H1 gemaggliutinina virusov grippa A/Kiev/59/79 i X/Leningrad/54/1 s pomoshch'iu oligosakharidnykh kart.

For comparative study of carbohydrate chains of N-glycoproteins, method of "oligosaccharide maps" has been developed. It consists in fractionation of reduced oligosaccharide fragments by gel-chromatography and HPLC on reverse phase and amino columns. Using two HPLC retention time values for each oligosaccharide, two-dimensional maps for both variants of H1 hemagglutinin were constructed. The monosaccharide composition of the majority of oligosaccharides isolated was also elucidated. The carbohydrate chain's patterns for the H1 hemagglutinin variants were found to be similar but to differ considerably from those for H3 hemagglutinin. The data obtained show that the glycosylation pattern depends on virus strain, i.e. on the structure of the polypeptide chain of hemagglutinin.

[Structure of major complex carbohydrate chains of influenza virus A/Leningrad/385/80 (H3N2) hemagglutinin]. / Stroenie glavnykh uglevodnykh tsepei kompleksnogo tipa gemaggliutinina virusa grippa A/Lenongrad/385/80 (H3N2).

The structure of four oligosaccharides which are the main carbohydrate chains of hemagglutinin of influenza virus A/Leningrad/385/80 (H3N2) has been elucidated. It was shown by means of enzymatic and mild acid hydrolysis, Smith degradation and acetolysis that the oligosaccharides have very similar structures (noncomplete triantennary) and differ from each other only in the number (0, 1 or 2) and position of fucose residues. The peculiarities of glycosylation of H3 hemagglutinin from different strains of influenza virus were discussed.

[Structure of major oligomannoside chains of influenza virus A/Leningrad/385/80 (H3 N2) hemagglutinin]. / Stroenie glavnykh oligomannozidnykh tsepei gemaggliutinina virusa grippa A/Leningrad/385/80 (H3 N2).

The structure of four main oligomannosidic carbohydrate chains isolated from influenza virus A/Leningrad/385/80 (H3N2) hemagglutinin has been elucidated using 1H NMR spectroscopy. The data obtained suggest that splitting off four alpha 1-2 linked mannose residues under alpha-mannosidase action is the limiting and selective stage of transformation of high mannose carbohydrate chain to complex chain during biosynthesis of glycoproteins.

[Heterogeneity of carbohydrate fragments in heavy and light chains of influenza virus A/Leningrad/385/80 (H3N2) hemagglutinin]. / Geterogennost' uglevodnykh fragmentov v tiazheloi i legkoi tsepiakh gemaggliutinina virusa grippa A/Leningrad/385/80 (H3N2).

Comparative analysis of carbohydrate chains variations in influenza virus A/Leningrad/385/80 (H3N2) hemagglutinin (HA) and its heavy (HA1) and light (HA2) chains has been carried out. The carbohydrate chains of these three glycoproteins were eliminated by reductive cleavage of N-glucosaminidic linkages under LiBH4 - tert-BuOH treatment. Fractionation of the oligosaccharides thus obtained by means of gel chromatography and HPLC resulted in isolation of 21 individual oligosaccharides from each glycoprotein. Their monosaccharide composition revealed almost identical pattern of high-mannose as well as complex chains in HA1 and HA2 in spite of different number (6-7 in HA1 and only 1 in HA2) of glycosilated sites. The possibility of a great number of both high-mannose and complex chains attached at the same site of glycoprotein is shown.

Some properties of cardiac troponin T structure.

Troponin T is eluted in multiple peaks when the whole bovine cardiac troponin complex is subjected to DEAE-cellulose chromatography in the presence of 8 M-urea. The heterogeneity observed is due to the presence of two forms of troponin T, differing in their Mr values, amino acid content, degree of phosphorylation and aggregation. Cardiac troponin T contains up to 0.8 mol of phosphate/mol of protein. Rabbit skeletal-muscle troponin T kinase phosphorylates the single site located in the N-terminal pentapeptide of cardiac troponin T. The composition of this peptide, (Ser,Asx,Glx,Glx)Val, is similar to that of skeletal-muscle troponin T. The single thiol group of cardiac troponin T is located some 50-70 residues from the N-terminus. The C-terminal sequence of cardiac troponin T is Trp-Lys, i.e. as is the case of skeletal-muscle troponin T.

[Structure of glyceraldehyde-3-phosphate dehydrogenase from rat muscle. Localization of arginine residues modified by 2,3-butanedione]. / Issledovanie struktury glitseral'degid-3-fosfatdegidrogenazy iz myshts krysy. Lokalizatsiia argininovykh ostatkov, modifitsiruemykh 2,3-butandionom.

Modification of rat skeletal muscle hologlyceraldehyde 3-phosphate dehydrogenase by [2,3-14C]butanedione was carried out. The conditions for obtaining preparative amounts of the modified protein and for the maintenance of the modification product stability at various steps of treatment were elaborated. Two radioactive peptides containing modified arginine residues were isolated from the trypsin hydrolysate of the enzyme modified by [2,3-14C] butanedione and oxidizied by performic acid, and their amino acid sequence was established. The data obtained suggest that the essential arginine residue occupies position 134 in the primary structure of the rat muscle enzyme.

[Serratia marcescens endonuclease. Properties of the enzyme]. / Endonukleaza Serratia marcescens. Kharakteristika fermenta.

Some physico-chemical properties of endonuclease (EC 3.1.4.9) from Serratia marcescens were studied and the amino acid composition of the enzyme was determined. The protein molecule was shown to contain one SH-group and one S-S-bond, which renders it different from the well studied nuclease (EC 3.1.4.7) from Staph. pyogenes. The conditions for reconstitution of the S-S-bond by dithioerythritol for quantitative estimation of cysteine residues of the endonuclease molecule were selected. The N-terminal amino acid was found to be threonine. The UV spectra for the enzyme are typical for proteins; A 0,1% 1cm,280nm is 1.46, epsilon 25 degrees 280nm,pH7,4 is 47292 M-1 cm-1. The sedimentation coefficient in phosphate buffer sW, 20 degrees is 3.4 S, pI is 6.5 and 7.5.