RESUMO
Bacterial capsular polysaccharides are important virulence factors. Capsular polysaccharides from several important Gram-negative pathogens share a conserved glycolipid terminus containing 3-deoxy-ß-d- manno-oct-2-ulosonic acid (ß-Kdo). The ß-Kdo glycosyltransferases responsible for synthesis of this conserved glycolipid belong to a new family of glycosyltransferases that shares little homology with other such enzymes, thereby representing an attractive antivirulence target. Here, we report the development of a fluorescence polarization-based, high-throughput screening assay (FP-tag) for ß-Kdo glycosyltransferases, and use it to identify a class of marine natural products as lead inhibitors. This "FP-tag" assay should be readily adaptable to high-throughput screens of other glycosyltransferases.
Assuntos
Inibidores Enzimáticos/farmacologia , Glicosiltransferases/antagonistas & inibidores , Química Click , Avaliação Pré-Clínica de Medicamentos , Inibidores Enzimáticos/química , Ensaios de Triagem em Larga Escala , Concentração Inibidora 50 , Açúcares Ácidos/química , Açúcares Ácidos/farmacologiaRESUMO
A convenient method for passivating and functionalizing stainless steel is described. Several methods of coating stainless steel (SS) samples with silica were investigated and of these methods, a thin (less than 15 nm thick) layer of silica created by atomic layer deposition (ALD) was found to give superior performance in electrochemical testing. These interfaces were then used as a platform for further functionalization with molecules of biological interest. Specifically, the SS samples were functionalized with biologically significant carbohydrates [N-acetyl-D-glucosamine (GlcNAc) and D-galactose (Gal)] that contain trialkoxysilane derivatives as chemical handles for linking to the surface. The presence and biological availability of these moieties on the silica coated SS were confirmed by XPS analysis and an enzyme-linked lectin assay (ELLA) using complementary lectins that specifically recognize the surface-bound carbohydrate. This method has the potential of being adapted to the functionalization of stainless steel biomedical implants with other biologically relevant carbohydrates.
Assuntos
Carboidratos/química , Materiais Revestidos Biocompatíveis/química , Aço Inoxidável/química , Polietilenoglicóis/química , Próteses e Implantes , Silanos/química , Dióxido de Silício/químicaRESUMO
Syntheses of buergerinin F (1) and buergerinin G (2) were carried out to establish the absolute stereochemistry of these natural products. A linear sequence was used to synthesize 1 in 15 steps and 9% overall yield from thymidine. Subsequent oxidation of 1 with ruthenium tetroxide afforded 2 in 77% yield.