sodium-dodecyl-sulfate and pyridine

Considering the importance of proteins in the structure and function of the cell wall of Candida albicans, we analyzed the cell wall subproteome of this important human pathogen by LC coupled to MS (LC-MS) using different protein extraction procedures. The analyzed samples included material extracted by hydrogen fluoride-pyridine (HF-pyridine), and whole SDS-extracted cell walls. The use of this latter innovative procedure gave similar data as compared to the analysis of HF-pyridine extracted proteins. A total of 21 cell wall proteins predicted to contain a signal peptide were identified, together with a high content of potentially glycosylated Ser/Thr residues, and the presence of a GPI motif in 19 of them. We also identified 66 "atypical" cell wall proteins that lack the above-mentioned characteristics. After tryptic removal of the most accessible proteins in the cell wall, several of the same expected GPI proteins and the most commonly found "atypical" wall proteins were identified. This result suggests that proteins are located not only at the cell wall surface, but are embedded within the cell wall itself. These results, which include new identified cell wall proteins, and comparison of proteins in blastospore and mycelial walls, will help to elucidate the C. albicans cell wall architecture.

Topics: Candida albicans; Cell Wall; Fungal Proteins; Hydrofluoric Acid; Proteome; Pyridines; Sodium Dodecyl Sulfate; Solvents; Tandem Mass Spectrometry

A monolithic capillary column was obtained by in-situ co-polymerization of butyl methacrylate with ethylene dimethacrylate. A mode of capillary electrochromatography (CEC) based on the dynamic adsorption of long chain sodium dodecyl sulfate(SDS) on the monolithic stationary phases was developed. Effects of SDS concentration and concentration of acetonitrile in mobile phase on the electroosmotic flow(EOF) of the column and the plate height on the electroosmotic velocity were investigated. Efficiency of monolithic capillary column with higher than 140,000 plates per meter for neutral aromatic compounds were obtained, and the relative standard deviations observed for dead time(t0) and retention times of neutral solutes were about 0.22% and less than 0.56% for ten consecutive runs, respectively.

Topics: Anions; Benzyl Alcohol; Chromatography, Micellar Electrokinetic Capillary; Electrochemistry; Electrophoresis, Capillary; Pyridines; Sodium Dodecyl Sulfate; Surface-Active Agents; Thiourea

A mode of capillary electrochromatography (CEC), based on the dynamical adsorption of surfactants on the uncharged monolithic stationary phases has been developed. The monolithic stationary phase, obtained by the in situ polymerization of butyl methacrylate with ethylene dimethacrylate, was dynamically modified with an ionic surfactant such as the long-chain quaternary ammonium salt of cetyltrimethylammonium bromide (CTAB) and long-chain sodium sulfate of sodium dodecyl sulfate (SDS). The ionic surfactant was adsorbed on the surface of polymeric monolith by hydrophobic interaction, and the ionic groups used to generate the electroosmotic flow (EOF). The electroosmotic mobility through these capillary columns increased with increasing the content of ionic surfactants in the mobile phase. In this way, the synthesis of the monolithic stationary phase with binary monomers can be controlled more easily than that with ternary monomers, one of which should be an ionic monomer to generate EOF. Furthermore, it is more convenient to change the direction and magnitude of EOF by changing the concentration of cationic or anionic surfactants in this system. An efficiency of monolithic capillary columns with more than 140000 plates per meter for neutral compounds has been obtained, and the relative standard deviations observed for to and retention factors of neutral solutes were about 0.22% and less than 0.56% for ten consecutive runs, respectively. Effects of mobile phase composition on the EOF of the column and the retention values of the neutral solutes were investigated. Simultaneous separation of basic, neutral and acidic compounds has been achieved.

Topics: Anions; Benzyl Alcohol; Cations; Cetrimonium; Cetrimonium Compounds; Electrophoresis, Capillary; Nitriles; Pyridines; Sodium Dodecyl Sulfate; Surface-Active Agents