Quantitative Proteome and Acidic Subproteome Profiling of Candida albicans Yeast-to-Hypha Transition

Abstract

Candida albicans yeast-to-hypha morphological transition is involved in the virulence strategy of this opportunistic fungal pathogen. Changes in relative abundance of the Candida proteome related to this process were analyzed using different two-dimensional differential in-gel electrophoresis (2D-DIGE)-based approaches. First, a comparative analysis of yeast and hyphal cytoplasmic proteins allowed the detection of 106 protein spots with significant variation in abundance. Sixty-one of them, corresponding to 46 proteins, were identified. As most of the differentially abundant proteins had an acidic isoelectric point, a large-scale prefractionation approach to analyze the acidic C. albicans subproteome was carried out. Ninety acidic C. albicans proteins were identified by either gel-based or nongel-based approaches. Additionally, different workflows combining preparative isoelectric focusing, Cy labeling, and narrow pH gradient 2-DE gels were tested to analyze the differences in relative protein abundance between yeast and hyphal acidic subproteomes. It was possible to identify 21 differentially abundant acidic proteins; 10 of them were not identified in the previous 2D-DIGE gels. Functional and network interaction analyses of the 56 differentially abundant proteins identified by both approaches rendered an integrated view of metabolic and cellular process reorganization during the yeast-to-hypha transition. With these results, we propose a model of metabolic reorganization.

Proteome changes during yeast-like and pseudohyphal growth in the biofilm-forming yeast Pichia fermentans

“…18S rRNA was used as reference gene Carbohydrate pathway Proteomic analysis showed a different organisation of the carbohydrate pathway in the two morphologies. Several proteins, belonging to glycolysis/gluoconeogenesis pathway, increased in abundance in yeast morphology of P. fermentans as a consequence of the exponential growth phase of the yeast form, and in accordance with data reported by Monteoliva et al (2011). This behaviour suggests that the yeast status is characterised by active aerobic glycolysis.…”

“…This behaviour might indicate that different species of the same protein might be involved in the dimorphic transition with different physiological relevance in the two morphologies (Jungblut et al 2008;Schlüter et al 2009). In Candida albicans, Monteoliva et al (2011) suggested that phosphoglycerate mutase can function as glycolytic enzyme providing energy for the yeast growth morphology, whereas in the filamentous growth, phosphoglycerate mutase might provide glucose donor for the synthesis of β-glucans and chitin. Further investigations should be performed to understand the function of different protein species in the transition of P. fermentans.…”

“…Additionally a proteomic approach could be also useful for elucidating the transition processes in P. fermentans. Proteomics has been successfully applied to study the effect of temperature on the proteome of P. pastoris (Dragosits et al 2009) and the yeast-to-hypha transition in Candida albicans (Monteoliva et al 2011). To the best of our knowledge, a study on the proteome changes in the two morphologies of P. fermentans has not yet been carried out.…”

Combining Proteomic Strategies and Molecular Display Technology for Development of Vaccines against Candida albicans

“…Other proteins in the FBS series showed moderate abundance changes [19]. Based on these results, we suggest that C. albicans employed the following adaptation strategy: first, C. albicans tuned its proteome to adapt to a new environment, in which most proteins were upregulated 2-fold higher than has been suggested by previous studies [23]. Second, a few proteins were upregulated >5-fold, which might suggest that these proteins were critical for adaptation to the novel environment.…”

Phosphoproteomic Analysis of Protein Kinase C Signaling in Saccharomyces cerevisiae Reveals Slt2 Mitogen-activated Protein Kinase (MAPK)-dependent Phosphorylation of Eisosome Core Components

“…In-solution digestion was performed as reported elsewhere (36). Proteins were reduced with 5 mM DTT for 30 min at 37°C and alkylated with 10 mM iodoacetamide for 30 min at 30°C.…”