Introducing fluorescence resonance energy transfer-based biosensors for the analysis of cAMP-PKA signalling in the fungal pathogen Candida glabrata.
Liesbeth DemuyserWouter Van GenechtenHideaki MizunoSonia ColomboPatrick Van DijckPublished in: Cellular microbiology (2018)
The cyclic adenosine monophosphate-protein kinase A (cAMP-PKA) pathway is central to signal transduction in many organisms. In pathogenic fungi such as Candida albicans, this signalling cascade has proven to be involved in several processes, such as virulence, indicating its potential importance in antifungal drug discovery. Candida glabrata is an upcoming pathogen of the same species, yet information regarding the role of cAMP-PKA signalling in virulence is largely lacking. To enable efficient monitoring of cAMP-PKA activity in this pathogen, we here present the usage of two FRET-based biosensors. Both variations in the activity of PKA and the quantity of cAMP can be detected in a time-resolved manner, as we exemplify by glucose-induced activation of the pathway. We also present information on how to adequately process and analyse the data in a mathematically correct and physiologically relevant manner. These sensors will be of great benefit for scientists interested in linking the cAMP-PKA signalling cascade to downstream processes, such as virulence, possibly in a host environment.
Keyphrases
- candida albicans
- biofilm formation
- energy transfer
- protein kinase
- binding protein
- drug discovery
- escherichia coli
- pseudomonas aeruginosa
- staphylococcus aureus
- quantum dots
- antimicrobial resistance
- single molecule
- health information
- healthcare
- type diabetes
- oxidative stress
- cystic fibrosis
- skeletal muscle
- endothelial cells
- metabolic syndrome
- big data
- electronic health record
- blood glucose
- living cells
- blood pressure