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Proteomic profile of Cryptococcus gattii biofilm: Metabolic shift and the potential activation of electron chain
Lucélia Santi1, Markus Berger2, Jorge A Guimarães3
1Faculty of Pharmacy, Federal University of Rio Grande do Sul, Porto Alegre, RS, Brazil; Post-Graduation Program of Cellular and Molecular Biology, Federal University of Rio Grande do Sul, Porto Alegre, RS, Brazil; Center of Experimental Research, Clinical Hospital of Porto Alegre, Porto Alegre, RS, Brazil.
Abstract:
Cryptococcus gattii is a primary pathogenic fungus that causes pneumonia. This species is also responsible for an outbreak in Vancouver, Canada, and spreading to the mainland and United States. The use of medical devices is often complicated by infections with biofilm-forming microbes with increased resistance to antimicrobial agents and host defense mechanisms. This study investigated the comparative proteome of C. gattii R265 (VGIIa) grown under planktonic and biofilm conditions. A brief comparison with C. neoformans H99 biofilm and the use of different culture medium and surface were also evaluated. Using Multidimensional Protein Identification Technology (MudPIT), 1819 proteins were identified for both conditions, where 150 (8.2%) were considered differentially regulated (up- or down-regulated and unique in biofilm cells). Overall, the proteomic approach suggests that C. gattii R265 biofilm cells are maintained by the induction of electron transport chain for reoxidation, and by alternative energy metabolites, such as succinate and acetate. SIGNIFICANCE: Since C. gattii is considered a primary pathogen and is one of the most virulent and less susceptible to antifungals, understanding how biofilms are maintained is fundamental to search for new targets to control this important mode of growth that is difficult to eradicate.
Insights
Cryptococcus gattii biofilms utilize an electron transport chain and alternative metabolites like succinate for maintenance. Understanding these mechanisms is key to combating this resilient fungal pathogen.
Area of Science:
- Mycology
- Proteomics
- Medical Microbiology
Background:
- Cryptococcus gattii is a primary fungal pathogen causing pneumonia.
- This fungus is responsible for outbreaks and exhibits increased resistance in biofilms.
- Biofilm formation complicates medical device use and antimicrobial treatment.
Purpose of the Study:
- To comparatively analyze the proteome of C. gattii R265 under planktonic and biofilm conditions.
- To investigate the role of electron transport and alternative metabolites in biofilm maintenance.
- To compare C. gattii biofilms with C. neoformans and evaluate culture conditions.
Main Methods:
- Multidimensional Protein Identification Technology (MudPIT) was employed.
- Proteomic analysis was performed on C. gattii R265 grown in planktonic and biofilm states.
- Comparative analysis included C. neoformans H99 and varied culture media/surfaces.
Main Results:
- 1819 proteins were identified in total.
- 150 proteins (8.2%) were differentially regulated in biofilm cells.
- Biofilm maintenance involves induced electron transport chain and utilization of succinate and acetate.
Conclusions:
- C. gattii biofilm cells are sustained through enhanced oxidative processes and alternative energy sources.
- Identifying targets to disrupt C. gattii biofilm formation is crucial for therapeutic strategies.
- This research provides insights into controlling a difficult-to-eradicate fungal growth mode.
