Sequential Fractionation and Two-dimensional Gel Analysis Unravels the Complexity of the

The cell wall proteins of Candida albicans play a key role in morphogenesis and pathogenesis and might be poten-tial target sites for new specific antifungal drugs. How-ever, these proteins are difficult to analyze because of their high heterogeneity, interconnections with wall po-lysaccharides (mannan, glucan, and chitin), low abun-dance, low solubility, and hydrophobic nature. Here we report a subproteomic approach for the study of the cell wall proteins (CWPs) from C. albicans yeast and hyphal forms. Most of the mannoproteins present in this com-partment were extracted by cell wall fractionation accord-ing to the type of interactions that they establish with other structural components. CWPs were solubilized from isolated cell walls by hot SDS and dithiothreitol treatment followed by extraction either by mild alkali conditions o