Structure elucidation and gene cluster characterization of the O-antigen of Yersinia kristensenii С-134.

Mild acid degradation of the lipopolysaccharide of Yersinia kristensenii C-134 afforded a glycerol teichoic acid-like O-polysaccharide, which was studied by sugar analysis, O-deacetylation and dephosphorylation along with 1D and 2D NMR spectroscopy. The following structure of the O-polysaccharide was established: This structure is related to those of other Y. kristensenii O-polysaccharides studied earlier. The O-antigen gene cluster of Y. kristensenii С-134 was analyzed and found to be consistent with the O-polysaccharide structure established.

Full structure and insight into the gene cluster of the O-specific polysaccharide of Yersinia intermedia H9-36/83 (O:17).

Lipopolysaccharide was isolated from bacteria Yersinia intermedia H9-36/83 (O:17) and degraded with mild acid to give an O-specific polysaccharide, which was isolated by GPC on Sephadex G-50 and studied by sugar analysis and 1D and 2D NMR spectroscopy. The polysaccharide was found to contain 3-deoxy-3-[(R)-3-hydroxybutanoylamino]-d-fucose (d-Fuc3NR3Hb) and the following structure of the heptasaccharide repeating unit was established: The structure established is consistent with the gene content of the O-antigen gene cluster. The O-polysaccharide structure and gene cluster of Y. intermedia are related to those of Hafnia alvei 1211 and Escherichia coli O:103.

Luminescent heterometallic gold-copper alkynyl complexes stabilized by tridentate phosphine.

The reactions between trinuclear gold complex tppmAu(3)Cl(3) (tppm = tris(diphenylphosphino)methane), arylacetylenes HC(2)C(6)H(4)X and Cu(+) under basic conditions result in formation of the heterometallic complexes [tppm(AuC(2)C(6)H(4)X)(3)Cu](+), X = H (1), COOMe (2), CN (3), OMe (4), NH(2) (5). These compounds belong to one structural motif and consist of the heterometallic {(AuC(2)C(6)H(4)X)(3)Cu} core stabilized by the tridentate phosphine. Compounds 1-5 were characterized by polynuclear NMR and IR spectroscopy, ESI-MS and single-crystal X-ray analysis. Luminescence properties of these complexes have been studied and revealed a substantial red shift of the emission maxima with the increase in the electron donicity of the alkynyl ligands substituents in the 550-680 nm range. The theoretical calculations of the electronic structures showed that variations of the substituents on the alkynyl ligands display very little effect on the molecular structural parameters but show appreciable influence on the orbital energies and luminescence characteristics of the compounds under study.

Probing elongating and branching ß-D-galactosyltransferase activities in Leishmania parasites by making use of synthetic phosphoglycans.

Protozoan parasites of the genus Leishmania synthesize lipophosphoglycans (LPGs), phosphoglycans and proteophosphoglycans that contain phosphosaccharide repeat units of [-6)Gal(ß1-4)Man(α1-OPO(3)H-]. The repeat structures are assembled by sequential addition of Manα1-OPO(3)H and ß-Gal. In this study, an UDP-Gal-dependent activity was detected in L. donovani and L. major membranes using synthetic phospho-oligosaccharide fragments of lipophosphoglycan as acceptor substrates. Incubation of a microsomal preparation from L. donovani or L. major parasites with synthetic substrates and UDP-[6-(3)H]Gal resulted in incorporation of radiolabel into these exogenous acceptors. The [(3)H]galactose-labeled products were characterized by degradation into radioactive, low molecular mass fragments upon hydrolysis with mild acid and treatment with ß-galactosidases. We showed that the activity detected with L. donovani membranes is the elongating ß-d-galactosyltransferase associated with LPG phosphosaccharide backbone biosynthesis (eGalT). The eGalT activity showed a requirement for the presence of at least one phosphodiester group in the substrate and it was enhanced dramatically when two or three phosphodiester groups were present. Using the same substrates we detected two types of galactosyltransferase activity in L. major membranes: the elongating ß-d-galactosyltransferase and a branching ß-d-galactosyltransferase (bGalT). Both L. major enzymes required a minimum of one phosphodiester group present in the substrate, but acceptors with two or three phosphodiester groups were found to be superior.

Synthesis of a quaternary polyprenyl ammonium salt.

Reaction of primary C(55)-allylic alcohol moraprenol (WT(3)C(7-9)-OH, a polyprenol from mulberry leaves) with triethylamine in the presence of phosphorus oxychloride leads to a quaternary ammonium chloride with a good yield (72%) and high cis-stereoselectivity of the terminal isoprene unit. Cationic polyprenyl derivatives may be useful for transfection and immunological studies.

Synthetic glycovaccine protects against the bite of leishmania-infected sand flies.

Leishmaniasis is a vectorborne disease transmitted to human and other mammalian hosts by sand fly bite. In the present study, we show that immunization with Leishmania mexicana promastigote secretory gel (PSG) or with a chemically defined synthetic glycovaccine containing the glycans found in L. mexicana PSG can provide significant protection against challenge by the bite of infected sand flies. Only the glycan from L. mexicana was protective; those from other species did not protect against L. mexicana infection. Furthermore, neither PSG nor the glycovaccine protected against artificial needle challenge, which is traditionally used in antileishmanial vaccine development. Conversely, an antigen preparation that was effective against needle challenge offered no protection against sand fly bite. These findings provide a new target for Leishmania vaccine development and demonstrate the critical role that the vector plays in the evaluation of candidate vaccines for leishmaniasis and other vectorborne diseases.

Parasite glycoconjugates. Part 16: Synthesis of a disaccharide and phosphorylated di- and tri-saccharides from Leishmania lipophosphoglycan.

A neutral disaccharide beta-D-Galp-(1-->4)-alpha-D-Manp and phosphorylated di- and tri-saccharides beta-D-Galp-(1-->3)-[H(2)PO(3)-6]-beta-D-Galp-O[CH(2)](8)CHCH(2) and beta-D-Galp-(1-->3)-[H(2)PO(3)-6]-beta-D-Galp-(1-->4)-alpha-D-Manp, which are fragments of the phosphoglycan portion of the surface lipophosphoglycan from Leishmania donovani (the disaccharide) or Leishmania major (all three compounds), were prepared and used as TLC standards to help the identification and differentiation of the elongating and branching beta-D-galactosyl transferase activities in Leishmania. The phosphosaccharides were synthesised using the H-phosphonate method for phosphorylation.

The structure and the Raman vibrational spectrum of the beryllium aquacation.

The experimental Raman vibrational spectrum of the 5.94 m water solution of the beryllium(II) chloride has been acquired. Theoretical frequencies, infrared and Raman intensities of the vibrational spectrum of the beryllium cation tetrahydrate have been calculated by means of quantum chemical approach. The peaks of the experimental spectrum have been assigned on the basis of the results of the quantum-chemical calculations. It has been shown that the hydrating surrounding of the aquacation increases effectively the frequency of the beryllium-oxygen stretching vibration by 16% in comparison with the free complex.

Synthetic fragments of antigenic lipophosphoglycans from Leishmania major and Leishmania mexicana and their use for characterisation of the Leishmania elongating alpha-D-mannopyranosylphosphate transferase.

The phosphorylated branched heptasaccharides 7 and 8, the octasaccharide 9 and the phosphorylated trisaccharides 5 and 6, which are fragments of the phosphoglycan portion of the surface lipophosphoglycans from Leishmania mexicana (5) or L. major (6-9), were synthesised by using the glycosyl hydrogenphosphonate method for the preparation of phosphodiester bridges. The compounds were tested as acceptor substrates/putative inhibitors for the Leishmania elongating alpha-D-mannosylphosphate transferase.