Interaction of Porin from Yersinia pseudotuberculosis with Different Structural Forms of Endogenous Lipopolysaccharide

The interaction of Yersinia pseudotuberculosis porin solubilized in deoxycholate with the S- and R-forms of endogenous lipopolysaccharide (LPS) was studied by the quenching of intrinsic protein fluorescence. The samples of S-LPS differed both in the length of O-specific polysaccharide (n = 1 and 4) and in the acylation degree of the 3-hydroxytetradecanoic acid residues of the lipid A moiety (12-66%). R-LPS (12%) binding to porin was found to occur with positive cooperativity on two integrated structural regions of the R-LPS macromolecule, namely, core oligosaccharide and lipid A. The mode of porin interaction with low-acylated S-LPSs (15 or 20%) coincided with a model involving three types of binding sites. The shape of Scatchard curves of binding indicates that a complex formation between porin and low-acylated S-LPS is cooperative at low and moderate ligand concentration, whereas at near-saturating LPS concentrations porin binds to LPS independently on two types of binding sites. The O-specific polysaccharide chain in the S-LPS macromolecule increases the affinity of its interaction with porin in comparison with R-LPS–porin binding. A significant increase (to 66%) in the degree of S-LPS acylation substantially changed its porin-binding character: the process becomes anti-cooperative with lowered affinity. Thus, the features of LPS–porin interaction significantly depend on the conformational changes in the LPS molecule due to expanding of its hydrophobic region.     

Structure and physiological activity of lemnan, Lemna minor L. pectin

A pectic polysaccharide, lemnan, was isolated from freshly collected duckweed Lemna minor L. Its sugar chain was shown to be mainly composed of the residues of D-galacturonic acid (64%), galactose, arabinose, xylose, and D-apiose, a branched chain sugar. The high content of D-apiose (25%) indicated that lemnan is an apiogalacturonan type pectin similar to zosteran, a pectic polysaccharide from a sea phanerogam of the Zosteraceae family. The results of partial acidic hydrolysis, pectinase digestion, and NMR studies of lemnan demonstrated that its macromolecule contains regions of the linear alpha-1,4-D-galacturonan and branched apiogalacturonan. The side chains of apiogalacturonan were found to be formed of single and 1,5-linked residues of D-apiofuranose attached to 2- and 3-positions of the D-galacturonic acid residues of the apiogalacturonan backbone. Lemnan was shown to exhibit an immunomodulatory effect by activating the system of phagocytosis.     

The isolation, preliminary study of structure and physiological activity of water-soluble polysaccharides from squeezed berries of Snowball tree Viburnum opulus

Water-soluble polysaccharide fractions VO1-VO4 were isolated from the squeezed berries of snowball tree (Viburnum opulus) by successive extraction with water at various temperatures and pH and with aqueous solutions of ammonium oxalate. These fractions were purified by ion-exchange chromatography on DEAE cellulose, and the homogeneity of the purified polysaccharides was determined by gel filtration on Sephacryl S-500. Acidic polysaccharides close to pectins in their sugar composition were found in all the extracts (fractions VO1-1, VO2-1, VO3-2, and VO4-2). Residues of galacturonic acid, galactose, arabinose, and (to a lesser extent) rhamnose are their main constituents. Neutral polysaccharides composed mainly of galactose and mannose residues were additionally found in fractions extracted with acidified water (pH 4.0) and with aqueous ammonium oxalate solutions. Partial acidic hydrolysis and digestion with pectinase of acidic polysaccharides indicated that their carbohydrate backbone consists of alpha-1,4-linked residues of D-galacturonic acid. NMR spectra of acidic polysaccharides (fractions VO3-2 and VO3-3) confirmed this and demonstrated that their side oligosaccharide chains are composed of beta-1,4-linked galactopyranose residues and of terminal and 2,5- and 3,5-substituted residues of alpha-arabinofuranose at a Gal: Ara ratio of 3:1. Some polysaccharides from V. opulus were found to possess an immunostimulating activity: they enhance phagocytosis, in particular, the phagocytic index and the secretion of lysosomal enzymes with peritoneal macrophages. Calcium ions were found to be necessary for the appearance of the stimulating effect of acidic polysaccharides from V. opulus.     

The effect of ultraviolet radiation on the growth and polysaccharide composition of a callus culture of Silene vulgaris

Ultraviolet radiation (wavelength, 280-315 nm; power, 0.2-13.0 W/m2; exposure, 1 or 3 h) was shown to change the growth of campion callus and the polysaccharide (pectin and arabinogalactan) composition of cell walls. An increase in the concentration of polysaccharides and a decrease in the content of arabinose and galactose residues in pectin and arabinogalactan were noted. For the majority of calluses, growth indices, specific growth rate, and biomass productivity (per 11 medium) were almost the same as in nonirradiated control cells. Maximum values of the growth index and specific growth rate, determined for dry biomass, were observed at a low dose of irradiation (0.2 W/m2) and an exposure of 3 h. A considerable decrease in the content of arabinose and galactose in pectin was noted at high doses of irradiation (exposure, 3 h). Samples of arabinogalactan were characterized by variable arabinose to galactose ratios, which were in the range 1 : (3.4-8.3).     

Capsular antigens of bacteria. Capsular antigens as the basis of vaccines against pathogenic bacteria

The role of bacterial capsular antigens represented in capsular polysaccharides and exoglycans in pathogenicity and virulence of bacteria is discussed in this review. Using capsular antigens for vaccines against severe diseases caused by capsular microorganisms is considered in detail. The use of conjugates of capsular polysaccharides and their fragments with proteins and peptides for vaccine as well as using liposomes as adjuvants for the capsular antigens are described. Data concerning structural elucidation of bacterial capsular antigens are given in the first part of this review. Published in Russian in Biokhimiya, 2006, Vol. 71, No. 9, pp. 1175–1182.     

Isolation of Polysaccharides from the Callus Culture of Lemna minor L.

Two fractions that included acid arabinogalactan and pectin were extracted from the callus culture of duckweed plants (Lemna minorL.) with water and ammonium oxalate. Residues of galactose and arabinose (ratio, (2.0–2.5) : 1) were the major constituents of acid arabinogalactan. The pectin fraction contained primarily residues of glycuronic acids, galactose, and arabinose. The percentages of arabinogalactan and pectin were similar. The yield of polysaccharide fractions did not depend on the method used for their isolation. Extraction with water, treatment of the biomass with aqueous formalin and dilute hydrochloric acid, and extraction with aqueous ammonium oxalate allowed us to obtain the pectin polysaccharide with the highest purity.     

Mitochondrial Phylogenomics of Modern and Ancient Equids

The genus Equus is richly represented in the fossil record, yet our understanding of taxonomic relationships within this genus remains limited. To estimate the phylogenetic relationships among modern horses, zebras, asses and donkeys, we generated the first data set including complete mitochondrial sequences from all seven extant lineages within the genus Equus. Bayesian and Maximum Likelihood phylogenetic inference confirms that zebras are monophyletic within the genus, and the Plains and Grevy’s zebras form a well-supported monophyletic group. Using ancient DNA techniques, we further characterize the complete mitochondrial genomes of three extinct equid lineages (the New World stilt-legged horses, NWSLH; the subgenus Sussemionus; and the Quagga, Equus quagga quagga). Comparisons with extant taxa confirm the NWSLH as being part of the caballines, and the Quagga and Plains zebras as being conspecific. However, the evolutionary relationships among the non-caballine lineages, including the now-extinct subgenus Sussemionus, remain unresolved, most likely due to extremely rapid radiation within this group. The closest living outgroups (rhinos and tapirs) were found to be too phylogenetically distant to calibrate reliable molecular clocks. Additional mitochondrial genome sequence data, including radiocarbon dated ancient equids, will be required before revisiting the exact timing of the lineage radiation leading up to modern equids, which for now were found to have possibly shared a common ancestor as far as up to 4 Million years ago (Mya).     

Effect of the rol genes from Agrobacterium Rhizogenes on the content and structure of pectic substances and glycanase activity in Rubia Cordifolia transgenic cell cultures

The expression of the rolB gene was found to increase the pectic yield in Rubia cordifolia cells, while the rolC gene inhibited the pectin production, which correlated with its expression level. The expression of the rolA, rolB, and rolC genes led to an increase in the content of arabinogalactan (AG) in cells. The increase in the expression of the rolB and rolC genes resulted in a more significant reduction in the content of arabinose residues in pectin, which was accompanied by an increased activity of α-L-arabinofuranosidase in cells. Moreover, the amount of galactose residues in pectin increased with the enhancement of the rolB expression due to a decrease in the activity of β-galactosidase in cells. The content of galacturonic acid residues in pectin from transgenic cultures decreased in the following order: rolC > rolB > rolA. The amount of arabinose residues in AG decreased independently of the gene type. The amount of arabinose residues in AG was found to be considerably reduced when the rolB expression level was increased.     

Structural study of bergenan,a pectin from <Emphasis Type="Italic">Bergenia crassifolia</Emphasis>

A pectin polysaccharide named bergenan was isolated from the freshly collected leaves of the leather bergenia Bergenia crassifolia by extraction with an aqueous solution of ammonium oxalate. The main component of its carbohydrate chain was shown to be the residues of D-galacturonic acid (about 80%). In addition, the polysaccharide contains the residues of galactose, arabinose, and rhamnose; their total content is less than 15%. It was shown that the bergenan samples from bergenia leaves collected at different vegetation periods (from July to September) do not substantially differ either in monosaccharide composition or in the viscosity of their aqueous solutions. The results of enzymatic hydrolysis by α-1,4-galacturonase (pectinase), partial acidic hydrolysis, NMR spectroscopy, and methylation with subsequent analysis of the results by GC-MS indicate that the bergenan macromolecule contains the regions of a linear α-1,4-D-galactopyranosyluronan and rhamnogalacturonan-I (RG-I). Galacturonan responds for a greater part of the macromolecule. A considerable amount of its constitutent galacturonic acid residues are present as methyl esters. The side chains in RG-I are attached to the rhamnopyranose residues of the backbone by a 1,4-linkages and are composed of the residues of terminal arabinofuranose and galactopyranose, 1,5-linked α-arabinofuranose, and 1,4- and 1,6-linked β-galactopyranose. The branching points of the side chains of the RG-I molecule are 3,4- and 3,6-di-O-substituted galactose residues.     

Pectin substances of the callus culture of Silene vulgaris (M.) G

Pectin-protein fraction SVC was isolated from the callus culture of the bladder campion (Silene vulgaris). The main components in it were residues of D-galacturonic acid, galactose, arabinose, rhamnose, and protein. Using ion-exchange chromatography, ultrafiltration, and acid and enzymatic hydrolysis, it was shown that SVC contained a mixture of molecules of linear pectin, branched pectin polysaccharide, and pectin-protein polymer. A fragment of the linear chain of galacturonan amounted to more than half of the entire carbohydrate silenan chain. The branched area of the macromolecule is represented by rhamnogalacturonan I. The pectin-protein polymer consisted mainly of protein and weakly branched pectin fragments with molecular mass of more than 300 kDa.