Studying submicrosecond protein folding kinetics using a photolabile caging strategy and time-resolved photoacoustic calorimetry.

Kinetic measurement of protein folding is limited by the method used to trigger folding. Traditional methods, such as stopped flow, have a long mixing dead time and cannot be used to monitor fast folding processes. Here, we report a compound, 4-(bromomethyl)-6,7-dimethoxycoumarin, that can be used as a "photolabile cage" to study the early stages of protein folding. The folding process of a protein, RD1, including kinetics, enthalpy, and volume change, was studied by the combined use of a phototriggered caging strategy and time-resolved photoacoustic calorimetry. The cage caused unfolding of the photolabile protein, and then a pulse UV laser (∼10(-9) s) was used to break the cage, leaving the protein free to refold and allowing the resolving of two folding events on a nanosecond time scale. This strategy is especially good for monitoring fast folding proteins that cannot be studied by traditional methods.

Characterization of Cell Wall Proteins in Saccharomyces cerevisiae Clinical Isolates Elucidates Hsp150p in Virulence.

The budding yeast Saccharomyces cerevisiae has recently been described as an emerging opportunistic fungal pathogen. Fungal cell wall mannoproteins have been demonstrated to be involved in adhesion to inert surfaces and might be engaged in virulence. In this study, we observed four clinical isolates of S. cerevisiae with relatively hydrophobic cell surfaces. Yeast cell wall subproteome was evaluated quantitatively by liquid chromatography/tandem mass spectrometry. We identified totally 25 cell wall proteins (CWPs) from log-phase cells, within which 15 CWPs were quantified. The abundance of Scw10p, Pst1p, and Hsp150p/Pir2p were at least 2 folds higher in the clinical isolates than in S288c lab strain. Hsp150p is one of the members in Pir family conserved in pathogenic fungi Candida glabrata and Candida albicans. Overexpression of Hsp150p in lab strain increased cell wall integrity and potentially enhanced the virulence of yeast. Altogether, these results demonstrated that quantitative cell wall subproteome was analyzed in clinical isolates of S. cerevisiae, and several CWPs, especially Hsp150p, were found to be expressed at higher levels which presumably contribute to strain virulence and fungal pathogenicity.

A novel chemical oxo-precipitation (COP) process for efficient remediation of boron wastewater at room temperature.

Chemical oxo-precipitation (COP), which combines treatment with an oxidant and precipitation using metal salts, was developed for treating boron-containing water under milder conditions (room temperature, pH 10) than those of conventional coagulation processes. The concentration of boron compounds was 1000mg-BL(-1). They included boric acid (H3BO3) and perborate (NaBO3). Precipitation using calcium chloride eliminated 80% of the boron from the perborate solution, but was unable to treat boric acid. COP uses hydrogen peroxide (H2O2) to pretreat boric acid, substantially increasing the removal of boron from boric acid solution by chemical precipitation from less than 5% to 80%. Furthermore, of alkaline earth metals, barium ions are the most efficient precipitant, and can increase the 80% boron removal to 98.5% at [H2O2]/[B] and [Ba]/[B] molar ratios of 2 and 1, respectively. The residual boron in the end water of COP contained 15ppm-B: this value cannot be achieved using conventional coagulation processes.